TY - JOUR AB - Background: The impact of surgical resection on the survival and functional outcomes of patients with multiple brain metastases remains a critical question in neuro-oncology. Methods: This retrospective study involved 160 patients who underwent surgical resection of brain metastases from 2017 to 2020. Patients were classified by the number of metastases—single, oligometastatic, or multiple—and whether complete removal of the main symptomatic lesion was achieved. Outcomes such as survival rates, complications, and functional status were assessed. Results: Among the patients, 48.1% had a single metastasis, 21.9% were oligometastatic, and 30% had multiple metastases. Survival did not differ by number of metastases when the main lesion was completely resected and remaining lesions were radiated (p = 0.6172). Complete resection increased mean survival to 15.74 months compared with 4.5 months without it. Additionally, patients who underwent complete resection experienced fewer seizures (16.2% vs. 32.6%, p = 0.019), implying a lower seizure risk. Functional independence was maintained post-surgery. Conclusions: While multiple brain metastases are generally associated with poor outcomes, a trend toward longer survival was observed after complete resection of the index metastasis, though this was not statistically significant. Radiation of residual lesions remains important to support prognosis. Reducing the tumor volume is key to lowering seizure risk. This study supports the role of aggressive surgical interventions, paired with radiation, to potentially enhance outcomes in patients with multiple brain metastases. AU - Goldberg, M.* AU - Kraus, L.M.* AU - Vatcheva, C.* AU - Bernhardt, D.* AU - Combs, S.E. AU - Negwer, C.* AU - Meyer, B.* AU - Wagner, A.* C1 - 75768 C2 - 57969 SP - 3281 - 3281 TI - Evaluating the impact of index metastasis resection in patients with multiple brain metastases. JO - Cancers VL - 17 IS - 20 PY - 2025 SN - 2072-6694 ER - TY - JOUR AB - OBJECTIVES: The present study aimed to compare the tumor growth delay between conventional radiotherapy (CRT) and the spatially fractionated modalities of microbeam radiation therapy (MRT) and minibeam radiation therapy (MBRT). In addition, we also determined the influence of beam width and the peak-to-valley dose ratio (PVDR) on tumor regrowth. METHODS: A549, a human non-small-cell lung cancer cell line, was implanted subcutaneously into the hind leg of female CD1-Foxn1nu mice. The animals were irradiated with sham, CRT, MRT, or MBRT. The spatially fractionated fields were created using two specially designed multislit collimators with a beam width of 50 μm and a center-to-center distance (CTC) of 400 μm for MRT and a beam width of 500 μm and 2000 μm CTC for MBRT. Additionally, the concept of the equivalent uniform dose (EUD) was chosen in our study. A dose of 20 Gy was applied to all groups with a PVDR of 20 for MBRT and MRT. Tumor growth was recorded until the tumors reached at least a volume that was at least three-fold of their initial value, and the growth delay was calculated. RESULTS: We saw a significant reduction in tumor regrowth following all radiation modalities. A growth delay of 11.1 ± 8 days was observed for CRT compared to the sham, whereas MBRT showed a delay of 20.2 ± 7.3 days. The most pronounced delay was observed in mice irradiated with MRT PVDR 20, with 34.9 ± 26.3 days of delay. CONCLUSIONS: The current study highlights the fact that MRT and MBRT modalities show a significant tumor growth delay in comparison to CRT at equivalent uniform doses. AU - Subramanian, N. AU - Čolić, A. AU - Santiago Franco, M. AU - Stolz, J. AU - Ahmed, M. AU - Bicher, S. AU - Winter, J. AU - Lindner, R. AU - Raulefs, S. AU - Combs, S.E. AU - Bartzsch, S. AU - Schmid, T.E. C1 - 73005 C2 - 56891 TI - Superior anti-tumor response after microbeam and minibeam radiation therapy in a lung cancer mouse model. JO - Cancers VL - 17 IS - 1 PY - 2025 SN - 2072-6694 ER - TY - JOUR AB - Microbeam radiation therapy (MRT) is a still pre-clinical form of spatially fractionated radiotherapy, which uses an array of micrometer-wide, planar beams of X-ray radiation. The dose modulation in MRT has proven effective in the treatment of tumors while being well tolerated by normal tissue. Research on understanding the underlying biological mechanisms mostly requires large third-generation synchrotrons. In this study, we aimed to develop a preclinical treatment environment that would allow MRT independent of synchrotrons. We built a compact microbeam setup for pre-clinical experiments within a small animal irradiator and present in vivo MRT application, including treatment planning, dosimetry, and animal positioning. The brain of an immobilized mouse was treated with MRT, excised, and immunohistochemically stained against γH2AX for DNA double-strand breaks. We developed a comprehensive treatment planning system by adjusting an existing dose calculation algorithm to our setup and attaching it to the open-source software 3D-Slicer. Predicted doses in treatment planning agreed within 10% with film dosimetry readings. We demonstrated the feasibility of MRT exposures in vivo at a compact source and showed that the microbeam pattern is observable in histological sections of a mouse brain. The platform developed in this study will be used for pre-clinical research of MRT. AU - Ahmed, M. AU - Bicher, S. AU - Combs, S.E. AU - Lindner, R. AU - Raulefs, S. AU - Schmid, T.E. AU - Spasova, S. AU - Stolz, J. AU - Wilkens, J.J.* AU - Winter, J. AU - Bartzsch, S. C1 - 69937 C2 - 55326 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - In vivo microbeam radiation therapy at a conventional small animal irradiator. JO - Cancers VL - 16 IS - 3 PB - Mdpi PY - 2024 SN - 2072-6694 ER - TY - JOUR AB - Objective: Radiotherapy, which is commonly used for the local control of thoracic cancers, also induces chronic inflammatory responses in the microvasculature of surrounding normal tissues such as the lung and heart that contribute to fatal radiation-induced lung diseases (RILDs) such as pneumonitis and fibrosis. In this study, we investigated the potential of cannabidiol (CBD) to attenuate the irradiation damage to the vasculature. Methods: We investigated the ability of CBD to protect a murine endothelial cell (EC) line (H5V) and primary lung ECs isolated from C57BL/6 mice from irradiation-induced damage in vitro and lung ECs (luECs) in vivo, by measuring the induction of oxidative stress, DNA damage, apoptosis (in vitro), and induction of inflammatory and pro-angiogenic markers (in vivo). Results: We demonstrated that a non-lethal dose of CBD reduces the irradiation-induced oxidative stress and early apoptosis of lung ECs by upregulating the expression of the cytoprotective mediator heme-oxygenase-1 (HO-1). The radiation-induced increased expression of inflammatory (ICAM-2, MCAM) and pro-angiogenic (VE-cadherin, Endoglin) markers was significantly reduced by a continuous daily treatment of C57BL/6 mice with CBD (i.p. 20 mg/kg body weight), 2 weeks before and 2 weeks after a partial irradiation of the lung (less than 20% of the lung volume) with 16 Gy. Conclusions: CBD has the potential to improve the clinical outcome of radiotherapy by reducing toxic side effects on the microvasculature of the lung. AU - Bauer, L.* AU - Alkotub, Ab. AU - Ballmann, M.* AU - Hasanzadeh Kafshgari, M.* AU - Rammes, G.* AU - Multhoff, G.* C1 - 72313 C2 - 56590 TI - Cannabidiol (CBD) protects lung endothelial cells from irradiation-induced oxidative stress and inflammation in vitro and in vivo. JO - Cancers VL - 16 IS - 21 PY - 2024 SN - 2072-6694 ER - TY - JOUR AB - Latin Americans have a rich genetic make-up that translates into heterogeneous fractions of the autosomal genome in runs of homozygosity (FROH) and heterogeneous types and proportions of indigenous American ancestry. While autozygosity has been linked to several human diseases, very little is known about the relationship between inbreeding, genetic ancestry, and cancer risk in Latin Americans. Chile has one of the highest incidences of gallbladder cancer (GBC) in the world, and we investigated the association between inbreeding, GBC, gallstone disease (GSD), and body mass index (BMI) in 4029 genetically admixed Chileans. We calculated individual FROH above 1.5 Mb and weighted polygenic risk scores for GSD, and applied multiple logistic regression to assess the association between homozygosity and GBC risk. We found that homozygosity was due to a heterogeneous mixture of genetic drift and consanguinity in the study population. Although we found no association between homozygosity and overall GBC risk, we detected interactions of FROH with sex, age, and genetic risk of GSD that affected GBC risk. Specifically, the increase in GBC risk per 1% FROH was 19% in men (p-value = 0.002), 30% in those under 60 years of age (p-value = 0.001), and 12% in those with a genetic risk of GSD above the median (p-value = 0.01). The present study highlighted the complex interplay between inbreeding, genetic ancestry, and genetic risk of GSD in the development of GBC. The applied methodology and our findings underscored the importance of considering the population-specific genetic architecture, along with sex- and age-specific effects, when investigating the genetic basis of complex traits in Latin Americans. AU - Ceballos, F.C.* AU - Boekstegers, F.* AU - Scherer, D.* AU - Barahona Ponce, C.* AU - Marcelain, K.* AU - Garate-Calderon, V.* AU - Waldenberger, M. AU - Morales, E.* AU - Rojas, A.* AU - Muñoz, C.* AU - Retamales, J.* AU - de Toro, G.* AU - Vera Kortmann, A.* AU - Barajas, O.* AU - Rivera, M.T.* AU - Cortes, A.* AU - Loader, D.* AU - Saavedra, J.* AU - Gutiérrez, L.* AU - Ortega, A.* AU - Bertrán, M.E.* AU - Bartolotti, L.* AU - Gabler, F.* AU - Campos, M.* AU - Alvarado, J.* AU - Moisán, F.* AU - Spencer, L.* AU - Nervi, B.* AU - Carvajal-Hausdorf, D.* AU - Losada, H.* AU - Almau, M.* AU - Fernandez, P.* AU - Olloquequi, J.* AU - Salinas, P.* AU - Lorenzo Bermejo, J.* C1 - 72939 C2 - 56789 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Inbreeding and gallbladder cancer risk: Homozygosity associations adjusted for indigenous american ancestry, BMI, and genetic risk of gallstone disease. JO - Cancers VL - 16 IS - 24 PB - Mdpi PY - 2024 SN - 2072-6694 ER - TY - JOUR AB - Our study aimed to identify predictors for the effectiveness of tumor regression in lung cancer patients undergoing neoadjuvant treatment and cancer resections. Patients admitted between 2016 and 2022 were included in the study. Based on the histology of the tumor, patients were categorized into a lung adenocarcinoma group (LUAD) and squamous cell carcinoma group (SQCA). Ninety-five patients with non-small-cell lung cancer were included in the study. A total of 58 (61.1%) and 37 (38.9%) patients were included in the LUAD and SQCA groups, respectively. Additionally, 9 (9.5%), 56 (58.9%), and 30 (31.6%) patients were categorized with a tumor regression score of I, II, and III, respectively. In multivariable analyses, histology of the primary tumor (SQCA), lymph node size in the preoperative CT scan (>1.7 cm), and absolute tumor size reduction after neoadjuvant treatment (>2.6 cm) independently predict effectiveness of tumor regression (OR [95% confidence interval, p-value] of 6.88 [2.40-19.77, p < 0.0001], 3.13 [1.11-8.83, p = 0.0310], and 3.76 [1.20-11.81, p = 0.0233], respectively). Age > 70 years, extended resection > one lobe, and tumor recurrence or metastasis were identified as significant independent predictors of reduced overall survival. Assessment of tumor size before and after neoadjuvant treatment might help to identify high-risk patients with decreased survival and to improve patient management and care. AU - Damirov, F.* AU - Stoleriu, M.-G. AU - Manapov, F. AU - Boedeker, E.* AU - Dreher, S.* AU - Gerz, S.* AU - Hehr, T.* AU - Sandner, E.* AU - Ott, G.* AU - Hatz, R. AU - Preissler, G. C1 - 71564 C2 - 56307 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Perioperative predictive factors for tumor regression and survival in non-Small cell lung cancer patients undergoing neoadjuvant treatment and lung resection. JO - Cancers VL - 16 IS - 16 PB - Mdpi PY - 2024 SN - 2072-6694 ER - TY - JOUR AB - GBM WHO CNS Grade 4 represents a major challenge for oncology due to its aggressive behavior. Conventional imaging has restrictions in detecting tumor recurrence. This prospective study aims to identify gene-based biomarkers in whole blood instead of isolating exosomes for the early detection of tumor recurrence. Blood samples (n = 33) were collected from seven GBM patients at time points before and after surgery as well as upon tumor recurrence. Four tumor tissue samples were assessed in parallel. Next-generation sequencing (NGS), including mRNA-seq and small RNA-seq, was used to analyze gene expression profiles in blood samples and tumor tissues. A novel filtering pipeline was invented to narrow down potential candidate genes. In total, between 6-93 mRNA and 1-19 small RNA candidates could be identified among the seven patients. The overlap of genes between the patients was minimal, indicating significant inter-individual variance among GBM patients. In summary, this prospective study supports the applicability of gene expression measurements in whole blood for the detection of tumor recurrence. It might provide an alternative to the challenging workflow of liquid biopsy after laborious exosome isolation from whole blood. AU - Muhtadi, R.* AU - Bernhardt, D.* AU - Multhoff, G.* AU - Hönikl, L.* AU - Combs, S.E. AU - Krieg, S.M.* AU - Gempt, J.* AU - Meyer, B.* AU - Barsegian, V.* AU - Lindemann, M.* AU - Kasper, M.* AU - Stewart, S.* AU - Port, M.* AU - Abend, M.* AU - Diehl, C. AU - Ostheim, P.* C1 - 71127 C2 - 56020 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Liquid biopsy in whole blood for identification of gene expression patterns (mRNA and miRNA) associated with recurrence of glioblastoma WHO CNS grade 4. JO - Cancers VL - 16 IS - 13 PB - Mdpi PY - 2024 SN - 2072-6694 ER - TY - JOUR AB - Chimeric antigen receptor-T cells have spearheaded the field of adoptive cell therapy and have shown remarkable results in treating hematological neoplasia. Because of the different biology of solid tumors compared to hematological tumors, response rates of CAR-T cells could not be transferred to solid entities yet. CAR engineering has added co-stimulatory domains, transgenic cytokines and switch receptors to improve performance and persistence in a hostile tumor microenvironment, but because of the inherent cell type limitations of CAR-T cells, including HLA incompatibility, toxicities (cytokine release syndrome, neurotoxicity) and high costs due to the logistically challenging preparation process for autologous cells, the use of alternative immune cells is gaining traction. NK cells and γδ T cells that do not need HLA compatibility or macrophages and dendritic cells with additional properties such as phagocytosis or antigen presentation are increasingly seen as cellular vehicles with potential for application. As these cells possess distinct properties, clinicians and researchers need a thorough understanding of their peculiarities and commonalities. This review will compare these different cell types and their specific modes of action seen upon CAR activation. AU - Nguyen, N.T.T.* AU - Müller, R.* AU - Briukhovetska, D.* AU - Weber, J.* AU - Feucht, J.* AU - Künkele, A.* AU - Hudecek, M.* AU - Kobold, S. C1 - 71310 C2 - 56066 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - The spectrum of CAR cellular effectors: Modes of action in anti-tumor immunity. JO - Cancers VL - 16 IS - 14 PB - Mdpi PY - 2024 SN - 2072-6694 ER - TY - JOUR AB - BACKGROUND/OBJECTIVES: Dysgeusia contributes to the derangement of nutritional status in patients with cancer as well as worsening the quality of life. There has been a lack of effective treatments for taste disorders provided by the pharmaceutical industry. METHODS: This was a pilot randomized, parallel, triple-blind, and placebo-controlled intervention clinical trial in which 31 malnourished patients with cancer and dysgeusia receiving antineoplastic treatment were randomized into three arms [standard dose of DMB (150 mg DMB/tablet), high dose of DMB (300 mg DMB/tablet) or placebo (300 mg freeze-dried strawberry)] for three months. Patients consumed a DMB or placebo tablet before each main meal. Using the nanopore methodology, we analyzed the oral microbiome of patients with cancer using saliva samples. RESULTS: All patients with cancer and dysgeusia had dysbiosis in terms of lower bacterial diversity and richness. DMB consumption was associated with changes in oral microbiome composition. Neither selected bacteria nor taste perception, type of diet, and cytokine levels were associated with mucositis. Likewise, alcohol and tobacco consumption as well as general and digestive toxicity due to systemic therapy were not associated with specific changes of the oral microbiome, according to logistic binary regression. The standard dose of DMB resulted in a lower abundance of Veillonella compared with the high DMB dose and placebo at 3 months after intervention with DMB. In particular, some species such as Streptococcus parasanguinis, Veillonella parvula, and Streptococcus mutans were less abundant in the DMB standard-dose group. Additionally, the consumption of a standard dose of DMB revealed a negative association between the concentrations of TNF-α and the abundance of species such as Streptococcus thermophilus, Streptococcus pneumoniae, Streptococcus dysgalactiae and Streptococcus agalactiae. CONCLUSIONS: Accordingly, regular DMB consumption could modify the oral microbiome in patients with cancer and dysgeusia, which may contribute to maintaining an appropriate immune response. However, as the present pilot study involved a small number of participants, further studies are necessary to draw robust conclusions from the data. AU - Plaza-Díaz, J.* AU - Ruiz Ojeda, F.J. AU - López-Plaza, B.* AU - Brandimonte-Hernández, M.* AU - Álvarez-Mercado, A.I.* AU - Arcos-Castellanos, L.* AU - Feliú-Batlle, J.* AU - Hummel, T.* AU - Palma-Milla, S.* AU - Gil, A.* C1 - 72028 C2 - 56552 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Effect of a novel food rich in miraculin on the oral microbiome of malnourished oncologic patients with dysgeusia. JO - Cancers VL - 16 IS - 19 PB - Mdpi PY - 2024 SN - 2072-6694 ER - TY - JOUR AB - Glioblastoma (GBM) constitutes the most common primary brain tumor and it remains incurable despite therapeutic advances. The high infiltration/invasion potential of GBM cells is considered to be one of the reasons for the inevitable recurrence of the disease. Radiotherapy (RT) is part of the standard care for patients with GBM, and its benefits on overall survival are extensively reported. However, numerous preclinical studies show that X-ray irradiation can enhance the motility of GBM cells. In the present review, we bring together state-of-the-art research on the impact of radiation on GBM cell motility. The mechanisms through which irradiation impacts the brain tumor microenvironment and the tumor cells themselves, leading to more aggressive/invasive tumors, are described. Finally, we summarize potential pharmacological strategies to overcome this problem. Clinical data validating the occurrence of these processes are urgently needed as they could be of great value for patient outcomes. With this comprehensive review, we expect to highlight the need for methods which allow for monitoring the post-irradiation invasive behavior of GBM in patients. AU - Santiago Franco, M. AU - Raulefs, S. AU - Schilling, D. AU - Combs, S.E. AU - Schmid, T.E. C1 - 72837 C2 - 56742 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Impact of radiation on invasion and migration of glioma in vitro and in vivo. JO - Cancers VL - 16 IS - 23 PB - Mdpi PY - 2024 SN - 2072-6694 ER - TY - JOUR AB - Background: Although the associations between genetic variations and lung cancer risk have been explored, the epigenetic consequences of DNA methylation in lung cancer development are largely unknown. Here, the genetically predicted DNA methylation markers associated with non–small cell lung cancer (NSCLC) risk by a two-stage case-control design were investigated. Methods: The genetic prediction models for methylation levels based on genetic and methylation data of 1595 subjects from the Framingham Heart Study were established. The prediction models were applied to a fixed-effect meta-analysis of screening data sets with 27,120 NSCLC cases and 27,355 controls to identify the methylation markers, which were then replicated in independent data sets with 7844 lung cancer cases and 421,224 controls. Also performed was a multi-omics functional annotation for the identified CpGs by integrating genomics, epigenomics, and transcriptomics and investigation of the potential regulation pathways. Results: Of the 29,894 CpG sites passing the quality control, 39 CpGs associated with NSCLC risk (Bonferroni-corrected p ≤ 1.67 × 10−6) were originally identified. Of these, 16 CpGs remained significant in the validation stage (Bonferroni-corrected p ≤ 1.28 × 10−3), including four novel CpGs. Multi-omics functional annotation showed nine of 16 CpGs were potentially functional biomarkers for NSCLC risk. Thirty-five genes within a 1-Mb window of 12 CpGs that might be involved in regulatory pathways of NSCLC risk were identified. Conclusions: Sixteen promising DNA methylation markers associated with NSCLC were identified. Changes of the methylation level at these CpGs might influence the development of NSCLC by regulating the expression of genes nearby. Plain Language Summary: The epigenetic consequences of DNA methylation in lung cancer development are still largely unknown. This study used summary data of large-scale genome-wide association studies to investigate the associations between genetically predicted levels of methylation biomarkers and non–small cell lung cancer risk at the first time. This study looked at how well larotrectinib worked in adult patients with sarcomas caused by TRK fusion proteins. These findings will provide a unique insight into the epigenetic susceptibility mechanisms of lung cancer. AU - Zhao, X.* AU - Yang, M.* AU - Fan, J.* AU - Wang, M.* AU - Wang, Y.* AU - Qin, N.* AU - Zhu, M.* AU - Jiang, Y.* AU - Gorlova, O.Y.* AU - Gorlov, I.P.* AU - Albanes, D.* AU - Lam, S.* AU - Tardón, A.* AU - Chen, C.* AU - Goodman, G.E.* AU - Bojesen, S.E.* AU - Landi, M.T.* AU - Johansson, M.* AU - Risch, A.* AU - Wichmann, H.-E. AU - Bickeböller, H.* AU - Christiani, D.C.* AU - Rennert, G.* AU - Arnold, S.M.* AU - Brennan, P.* AU - Field, J.K.* AU - Shete, S.* AU - Le Marchand, L.* AU - Liu, G.* AU - Hung, R.J.* AU - Andrew, A.S.* AU - Kiemeney, L.A.* AU - Zienolddiny, S.* AU - Grankvist, K.* AU - Caporaso, N.E.* AU - Woll, P.J.* AU - Lazarus, P.* AU - Schabath, M.B.* AU - Aldrich, M.C.* AU - Patel, A.V.* AU - Davies, M.P.A.* AU - Ma, H.* AU - Jin, G.* AU - Hu, Z.* AU - Amos, C.I.* AU - Shen, H.* AU - Dai, J.* C1 - 68923 C2 - 53677 CY - 111 River St, Hoboken 07030-5774, Nj Usa SP - 913–926 TI - Identification of genetically predicted DNA methylation markers associated with non–small cell lung cancer risk among 34,964 cases and 448,579 controls. JO - Cancers VL - 130 PB - Wiley PY - 2024 SN - 2072-6694 ER - TY - JOUR AB - Historically, CD8+ T cells have been considered the most relevant effector cells involved in the immune response against tumors and have therefore been the focus of most cancer immunotherapy approaches. However, CD4+ T cells and their secreted factors also play a crucial role in the tumor microenvironment and can orchestrate both pro- and antitumoral immune responses. Depending on the cytokine milieu to which they are exposed, CD4+ T cells can differentiate into several phenotypically different subsets with very divergent effects on tumor progression. In this review, we provide an overview of the current knowledge about the role of the different T helper subsets in the immune system, with special emphasis on their implication in antitumoral immune responses. Furthermore, we also summarize therapeutic applications of each subset and its associated cytokines in the adoptive cell therapy of cancer. AU - Andreu-Sanz, D.* AU - Kobold, S. C1 - 67654 C2 - 53961 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Role and potential of different T helper cell subsets in adoptive cell therapy. JO - Cancers VL - 15 IS - 6 PB - Mdpi PY - 2023 SN - 2072-6694 ER - TY - JOUR AB - Recent epidemiologic studies support an association between chronic low-dose radiation exposure and the development of cardiovascular disease (CVD). The molecular mechanisms underlying the adverse effect of chronic low dose exposure are not fully understood. To address this issue, we have investigated changes in the heart proteome of ApoE deficient (ApoE-/-) C57Bl/6 female mice chronically irradiated for 300 days at a very low dose rate (1 mGy/day) or at a low dose rate (20 mGy/day), resulting in cumulative whole-body doses of 0.3 Gy or 6.0 Gy, respectively. The heart proteomes were compared to those of age-matched sham-irradiated ApoE-/- mice using label-free quantitative proteomics. Radiation-induced proteome changes were further validated using immunoblotting, enzyme activity assays, immunohistochemistry or targeted transcriptomics. The analyses showed persistent alterations in the cardiac proteome at both dose rates; however, the effect was more pronounced following higher dose rates. The altered proteins were involved in cardiac energy metabolism, ECM remodelling, oxidative stress, and ageing signalling pathways. The changes in PPARα, SIRT, AMPK, and mTOR signalling pathways were found at both dose rates and in a dose-dependent manner, whereas more changes in glycolysis and ECM remodelling were detected at the lower dose rate. These data provide strong evidence for the possible risk of cardiac injury following chronic low dose irradiation and show that several affected pathways following chronic irradiation overlap with those of ageing-associated heart pathology. AU - Azimzadeh, O.* AU - Merl-Pham, J. AU - Subramanian, V.* AU - Oleksenko, K. AU - Krumm, F.* AU - Mancuso, M.* AU - Pasquali, E.* AU - Tanaka, I.B.* AU - Tanaka, S.* AU - Atkinson, M.J. AU - Tapio, S. AU - Moertl, S.* C1 - 68106 C2 - 54584 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Late effects of chronic low dose rate total body irradiation on the heart proteome of ApoE-/- mice resemble premature cardiac ageing. JO - Cancers VL - 15 IS - 13 PB - Mdpi PY - 2023 SN - 2072-6694 ER - TY - JOUR AB - PURPOSE: Identification of molecularly-defined cancer subgroups and targeting tumor-specific vulnerabilities have a strong potential to improve treatment response and patient outcomes but remain an unmet challenge of high clinical relevance, especially in head and neck squamous cell carcinoma (HNSC). EXPERIMENTAL DESIGN: We established a UCHL1-related gene set to identify and molecularly characterize a UCHL1-related subgroup within TCGA-HNSC by integrative analysis of multi-omics data. An extreme gradient boosting model was trained on TCGA-HNSC based on GSVA scores for gene sets of the MSigDB to robustly predict UCHL1-related cancers in other solid tumors and cancer cell lines derived thereof. Potential vulnerabilities of UCHL1-related cancer cells were elucidated by an in-silico drug screening approach. RESULTS: We established a 497-gene set, which stratified the TCGA-HNSC cohort into distinct subgroups with a UCHL1-related or other phenotype. UCHL1-related HNSC were characterized by higher frequencies of genomic alterations, which was also evident for UCHL1-related cancers of other solid tumors predicted by the classification model. These data indicated an impaired maintenance of genomic integrity and vulnerability for DNA-damaging treatment, which was supported by a favorable prognosis of UCHL1-related tumors after radiotherapy, and a higher sensitivity of UCHL1-related cancer cells to irradiation or DNA-damaging compounds (e.g., Oxaliplatin). CONCLUSION: Our study established UCHL1-related cancers as a novel subgroup across most solid tumor entities with a unique molecular phenotype and DNA-damaging treatment as a specific vulnerability, which requires further proof-of-concept in pre-clinical models and future clinical trials. AU - Burkart, S.* AU - Weusthof, C.* AU - Khorani, K.* AU - Steen, S.* AU - Stögbauer, F.* AU - Unger, K. AU - Hess J. AU - Zitzelsberger, H. AU - Belka, C. AU - Kurth, I.* AU - Hess, J.* C1 - 67653 C2 - 53960 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - A novel subgroup of UCHL1-related cancers is associated with genomic instability and sensitivity to DNA-damaging treatment. JO - Cancers VL - 15 IS - 6 PB - Mdpi PY - 2023 SN - 2072-6694 ER - TY - JOUR AB - BACKGROUND: Resection of brain metastases (BM) close to motor structures is challenging for treatment. Navigated transcranial magnetic stimulation (nTMS) motor mapping, combined with diffusion tensor imaging (DTI)-based fiber tracking (DTI-FTmot.TMS), is a valuable tool in neurosurgery to preserve motor function. This study aimed to assess the practicability of DTI-FTmot.TMS for local adjuvant radiotherapy (RT) planning of BM. METHODS: Presurgically generated DTI-FTmot.TMS-based corticospinal tract (CST) reconstructions (FTmot.TMS) of 24 patients with 25 BM resected during later surgery were incorporated into the RT planning system. Completed fractionated stereotactic intensity-modulated RT (IMRT) plans were retrospectively analyzed and adapted to preserve FTmot.TMS. RESULTS: In regular plans, mean dose (Dmean) of complete FTmot.TMS was 5.2 ± 2.4 Gy. Regarding planning risk volume (PRV-FTTMS) portions outside of the planning target volume (PTV) within the 17.5 Gy (50%) isodose line, the DTI-FTmot.TMS Dmean was significantly reduced by 33.0% (range, 5.9-57.6%) from 23.4 ± 3.3 Gy to 15.9 ± 4.7 Gy (p < 0.001). There was no significant decline in the effective treatment dose, with PTV Dmean 35.6 ± 0.9 Gy vs. 36.0 ± 1.2 Gy (p = 0.063) after adaption. CONCLUSIONS: The DTI-FTmot.TMS-based CST reconstructions could be implemented in adjuvant IMRT planning of BM. A significant dose reduction regarding motor structures within critical dose levels seems possible. AU - Diehl, C. AU - Rosenkranz, E.* AU - Schwendner, M.J.* AU - Mißlbeck, M.* AU - Sollmann, N.* AU - Ille, S.* AU - Meyer, B.* AU - Combs, S.E. AU - Krieg, S.M.* C1 - 67176 C2 - 53513 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Dose reduction to motor structures in adjuvant fractionated stereotactic radiotherapy of brain metastases: nTMS-derived DTI-based motor fiber tracking in treatment planning. JO - Cancers VL - 15 IS - 1 PB - Mdpi PY - 2023 SN - 2072-6694 ER - TY - JOUR AB - BACKGROUND: Resection followed by local radiation therapy (RT) is the standard of care for symptomatic brain metastases. However, the optimal technique, fractionation scheme and dose are still being debated. Lately, low-energy X-ray intraoperative RT (lex-IORT) has been of increasing interest. METHOD: Eighteen consecutive patients undergoing BM resection followed by immediate lex-IORT with 16-30 Gy applied to the spherical applicator were retrospectively analyzed. Demographic, RT-specific, radiographic and clinical data were reviewed to evaluate the effectiveness and safety of IORT for BM. Descriptive statistics and Kaplan-Meyer analysis were applied. RESULTS: The mean follow-up time was 10.8 months (range, 0-39 months). The estimated local control (LC), distant brain control (DBC) and overall survival (OS) at 12 months post IORT were 92.9% (95%-CI 79.3-100%), 71.4% (95%-CI 50.2-92.6%) and 58.0% (95%-CI 34.1-81.9%), respectively. Two patients developed radiation necrosis (11.1%) and wound infection (CTCAE grade III); both had additional adjuvant treatment after IORT. For five patients (27.8%), the time to the start or continuation of systemic treatment was ≤15 days and hence shorter than wound healing and adjuvant RT would have required. CONCLUSION: In accordance with previous series, this study demonstrates the effectiveness and safety of IORT in the management of brain metastases despite the small cohort and the retrospective characteristic of this analysis. AU - Diehl, C. AU - Pigorsch, S.U. AU - Gempt, J.* AU - Krieg, S.M.* AU - Reitz, S.* AU - Waltenberger, M.* AU - Barz, M.* AU - Meyer, H.S.* AU - Wagner, A.* AU - Wilkens, J.* AU - Wiestler, B.* AU - Zimmer, C.* AU - Meyer, B.* AU - Combs, S.E. C1 - 67177 C2 - 53516 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Low-energy X-ray intraoperative radiation therapy (Lex-IORT) for resected brain metastases: A single-institution experience. JO - Cancers VL - 15 IS - 1 PB - Mdpi PY - 2023 SN - 2072-6694 ER - TY - JOUR AB - Postsurgical radiotherapy (RT) has been early proven to prevent local tumor recurrence, initially performed with whole brain RT (WBRT). Subsequent to disadvantageous cognitive sequalae for the patient and the broad distribution of modern linear accelerators, focal irradiation of the tumor has omitted WBRT in most cases. In many studies, the effectiveness of local RT of the resection cavity, either as single-fraction stereotactic radiosurgery (SRS) or hypo-fractionated stereotactic RT (hFSRT), has been demonstrated to be effective and safe. However, whereas prospective high-level incidence is still lacking on which dose and fractionation scheme is the best choice for the patient, further ablative techniques have come into play. Neoadjuvant SRS (N-SRS) prior to resection combines straightforward target delineation with an accelerated post-surgical phase, allowing an earlier start of systemic treatment or rehabilitation as indicated. In addition, low-energy intraoperative RT (IORT) on the surgical bed has been introduced as another alternative to external beam RT, offering sterilization of the cavity surface with steep dose gradients towards the healthy brain. This consensus paper summarizes current local treatment strategies for resectable brain metastases regarding available data and patient-centered decision-making. AU - Diehl, C. AU - Giordano, F.A.* AU - Grosu, A.L.* AU - Ille, S.* AU - Kahl, K.H.* AU - Onken, J.* AU - Rieken, S.* AU - Sarria, G.R.* AU - Shiban, E.* AU - Wagner, A.* AU - Beck, J.* AU - Brehmer, S.* AU - Ganslandt, O.* AU - Hamed, M.* AU - Meyer, B.* AU - Munter, M.* AU - Raabe, A.* AU - Rohde, V.* AU - Schaller, K.* AU - Schilling, D. AU - Schneider, M.* AU - Sperk, E.* AU - Thomé, C.* AU - Vajkoczy, P.* AU - Vatter, H.* AU - Combs, S.E. C1 - 68053 C2 - 54531 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Opportunities and alternatives of modern radiation oncology and surgery for the management of resectable brain metastases. JO - Cancers VL - 15 IS - 14 PB - Mdpi PY - 2023 SN - 2072-6694 ER - TY - JOUR AB - BACKGROUND: The aim of this study was to develop and validate radiogenomic models to predict the MDM2 gene amplification status and differentiate between ALTs and lipomas on preoperative MR images. METHODS: MR images were obtained in 257 patients diagnosed with ALTs (n = 65) or lipomas (n = 192) using histology and the MDM2 gene analysis as a reference standard. The protocols included T2-, T1-, and fat-suppressed contrast-enhanced T1-weighted sequences. Additionally, 50 patients were obtained from a different hospital for external testing. Radiomic features were selected using mRMR. Using repeated nested cross-validation, the machine-learning models were trained on radiomic features and demographic information. For comparison, the external test set was evaluated by three radiology residents and one attending radiologist. RESULTS: A LASSO classifier trained on radiomic features from all sequences performed best, with an AUC of 0.88, 70% sensitivity, 81% specificity, and 76% accuracy. In comparison, the radiology residents achieved 60-70% accuracy, 55-80% sensitivity, and 63-77% specificity, while the attending radiologist achieved 90% accuracy, 96% sensitivity, and 87% specificity. CONCLUSION: A radiogenomic model combining features from multiple MR sequences showed the best performance in predicting the MDM2 gene amplification status. The model showed a higher accuracy compared to the radiology residents, though lower compared to the attending radiologist. AU - Foreman, S.C.* AU - Llorián-Salvador, O.* AU - David, D.E.* AU - Rösner, V.K.N.* AU - Rischewski, J.F.* AU - Feuerriegel, G.C.* AU - Kramp, D.W.* AU - Luiken, I.* AU - Lohse, A.K.* AU - Kiefer, J.* AU - Mogler, C.* AU - Knebel, C.* AU - Jung, M.* AU - Andrade-Navarro, M.A.* AU - Rost, B.* AU - Combs, S.* AU - Makowski, M.R.* AU - Woertler, K.* AU - Peeken, J.C. AU - Gersing, A.S.* C1 - 67614 C2 - 53921 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Development and Evaluation of MR-Based Radiogenomic Models to Differentiate Atypical Lipomatous Tumors from Lipomas. JO - Cancers VL - 15 IS - 7 PB - Mdpi PY - 2023 SN - 2072-6694 ER - TY - JOUR AB - Ewing sarcoma (EwS) is a rare and predominantly pediatric malignancy of bone and soft tissue in children and adolescents. Although international collaborations have greatly improved the prognosis of most EwS, the occurrence of macrometastases or relapse remains challenging. The prototypic oncogene EWS-FLI1 acts as an aberrant transcription factor that drives the cellular transformation of EwS. In addition to its involvement in RNA splicing and the DNA damage response, this chimeric protein directly binds to GGAA repeats, thereby modifying the transcriptional profile of EwS. Direct pharmacological targeting of EWS-FLI1 is difficult because of its intrinsically disordered structure. However, targeting the EWS-FLI1 protein complex or downstream pathways provides additional therapeutic options. This review describes the EWS-FLI1 protein partners and downstream pathways, as well as the related target therapies for the treatment of EwS. AU - Gong, H.* AU - Xue, B.* AU - Ru, J. AU - Pei, G.* AU - Li, Y.* C1 - 67949 C2 - 54427 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Targeted therapy for EWS-FLI1 in ewing sarcoma. JO - Cancers VL - 15 IS - 16 PB - Mdpi PY - 2023 SN - 2072-6694 ER - TY - JOUR AB - Pancreatic ductal adenocarcinoma (PDAC) ranks among the most fatal cancer diseases, widely accepted to have the most dismal prognoses. Although immunotherapy has broadly revolutionized cancer treatment, its value in PDAC appears to be relatively low. Exhibiting protumoral effects, monocytes have recently been proposed as potential targets of such immunotherapeutic regimens. However, to date, the body of evidence on monocytes' role in PDAC is scarce. Therefore, we analyzed monocytes in the peripheral blood of 58 PDAC patients prior to surgery and compared them to healthy individuals. PDAC patients showed increased levels of monocytes when compared to healthy controls In addition, patients with perineural infiltration demonstrated a higher percentage of monocytes compared to non-infiltrating tumors and PDAC G3 was associated with higher monocyte levels than PDAC G2. Patients with monocyte levels > 5% were found to have an 8.9-fold increased risk for a G3 and perineural infiltrated PDAC resulting in poorer survival compared to patients with <5% monocyte levels. Furthermore, PDAC patients showed increased expressions of CD86 and CD11c and decreased expressions of PD-L1 on monocytes compared to healthy individuals. Finally, levels of monocytes correlated positively with concentrations of IL-6 and TNF-α in plasma of PDAC patients. Based on our findings, we propose monocytes as a novel prognostic biomarker. Large-scale studies are needed to further decipher the role of monocytes in PDAC and investigate their potential as therapeutic targets. AU - Hansen, F.J.* AU - David, P.* AU - Akram, M.* AU - Knoedler, S. AU - Mittelstädt, A.* AU - Merkel, S.* AU - Podolska, M.J.* AU - Swierzy, I.* AU - Roßdeutsch, L.* AU - Klösch, B.* AU - Kouhestani, D.* AU - Anthuber, A.* AU - Bénard, A.* AU - Brunner, M.* AU - Krautz, C.* AU - Grützmann, R.* AU - Weber, G.F.* C1 - 67266 C2 - 54203 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Circulating monocytes serve as novel prognostic biomarker in pancreatic ductal adenocarcinoma patients. JO - Cancers VL - 15 IS - 2 PB - Mdpi PY - 2023 SN - 2072-6694 ER - TY - JOUR AB - BACKGROUND: Despite advances in treatment for brain metastases (BMs), the prognosis for recurrent BMs remains poor and requires further research to advance clinical management and improve patient outcomes. In particular, data addressing the impact of tumor volume and surgical resection with regard to survival remain scarce. METHODS: Adult patients with recurrent BMs between December 2007 and December 2022 were analyzed. A distinction was made between operated and non-operated patients, and the residual tumor burden (RTB) was determined by using (postoperative) MRI. Survival analysis was performed and RTB cutoff values were calculated using maximally selected log-rank statistics. In addition, further analyses on systemic tumor progression and (postoperative) tumor therapy were conducted. RESULTS: In total, 219 patients were included in the analysis. Median age was 60 years (IQR 52-69). Median preoperative tumor burden was 2.4 cm3 (IQR 0.8-8.3), and postoperative tumor burden was 0.5 cm3 (IQR 0.0-2.9). A total of 95 patients (43.4%) underwent surgery, and complete cytoreduction was achieved in 55 (25.1%) patients. Median overall survival was 6 months (IQR 2-10). Cutoff RTB in all patients was 0.12 cm3, showing a significant difference (p = 0.00029) in overall survival (OS). Multivariate analysis showed preoperative KPSS (HR 0.983, 95% CI, 0.967-0.997, p = 0.015), postoperative tumor burden (HR 1.03, 95% CI 1.008-1.053, p = 0.007), and complete vs. incomplete resection (HR 0.629, 95% CI 0.420-0.941, p = 0.024) as significant. Longer survival was significantly associated with surgery for recurrent BMs (p = 0.00097), and additional analysis demonstrated the significant effect of complete resection on survival (p = 0.0027). In the subgroup of patients with systemic progression, a cutoff RTB of 0.97 cm3 (p = 0.00068) was found; patients who had received surgery also showed prolonged OS (p = 0.036). Single systemic therapy (p = 0.048) and the combination of radiotherapy and systemic therapy had a significant influence on survival (p = 0.036). CONCLUSIONS: RTB is a strong prognostic factor for survival in patients with recurrent BMs. Operated patients with recurrent BMs showed longer survival independent of systemic progression. Maximal cytoreduction should be targeted to achieve better long-term outcomes. AU - Lin, J.* AU - Kaiser, Y.* AU - Wiestler, B.* AU - Bernhardt, D.* AU - Combs, S.E. AU - Delbridge, C.* AU - Meyer, B.* AU - Gempt, J.* AU - Aftahy, A.K.* C1 - 68688 C2 - 54897 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Cytoreduction of residual tumor burden is decisive for prolonged survival in patients with recurrent brain metastases-retrospective analysis of 219 patients. JO - Cancers VL - 15 IS - 20 PB - Mdpi PY - 2023 SN - 2072-6694 ER - TY - JOUR AB - Somatostatin receptor (SSTR) agonists have been extensively used for treating neuroendocrine tumors. Synthetic therapeutic agonists showing selectivity for SSTR2 (Octreotide) or for SSTR2 and SSTR5 (Pasireotide) have been approved for the treatment of patients with acromegaly and Cushing's syndrome, as their pituitary tumors highly express SSTR2 or SSTR2/SSTR5, respectively. Nonfunctioning pituitary adenomas (NFPAs), which express high levels of SSTR3 and show only modest response to currently available SSTR agonists, are often invasive and cannot be completely resected, and therefore easily recur. The aim of the present study was the evaluation of ITF2984, a somatostatin analog and full SSTR3 agonist, as a new potential treatment for NFPAs. ITF2984 shows a 10-fold improved affinity for SSTR3 compared to Octreotide or Pasireotide. Molecular modeling and NMR studies indicated that the higher affinity for SSTR3 correlates with a higher stability of a distorted β-I turn in the cyclic peptide backbone. ITF2984 induces receptor internalization and phosphorylation, and triggers G-protein signaling at pharmacologically relevant concentrations. Furthermore, ITF2984 displays antitumor activity that is dependent on SSTR3 expression levels in the MENX (homozygous mutant) NFPA rat model, which closely recapitulates human disease. Therefore, ITF2984 may represent a novel therapeutic option for patients affected by NFPA. AU - Modena, D.* AU - Moras, M.L.* AU - Sandrone, G.* AU - Stevenazzi, A.* AU - Vergani, B.* AU - Dasgupta, P.* AU - Kliever, A.* AU - Gulde, S. AU - Marangelo, A. AU - Schillmaier, M.* AU - Luque, R.M.* AU - Bäuerle, S.* AU - Pellegata, N.S. AU - Schulz, S.* AU - Steinkühler, C.* C1 - 68105 C2 - 54583 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Identification of a Novel SSTR3 full agonist for the treatment of nonfunctioning pituitary adenomas. JO - Cancers VL - 15 IS - 13 PB - Mdpi PY - 2023 SN - 2072-6694 ER - TY - JOUR AB - Kirsten rat sarcoma virus (KRAS)-mutant cancers are frequent, metastatic, lethal, and largely undruggable. While interleukin (IL)-1β and nuclear factor (NF)-κB inhibition hold promise against cancer, untargeted treatments are not effective. Here, we show that human KRAS-mutant cancers are addicted to IL-1β via inflammatory versican signaling to macrophage inhibitor of NF-κB kinase (IKK) β. Human pan-cancer and experimental NF-κB reporter, transcriptome, and proteome screens reveal that KRAS-mutant tumors trigger macrophage IKKβ activation and IL-1β release via secretory versican. Tumor-specific versican silencing and macrophage-restricted IKKβ deletion prevents myeloid NF-κB activation and metastasis. Versican and IKKβ are mutually addicted and/or overexpressed in human cancers and possess diagnostic and prognostic power. Non-oncogene KRAS/IL-1β addiction is abolished by IL-1β and TLR1/2 inhibition, indicating cardinal and actionable roles for versican and IKKβ in metastasis. AU - Spella, M. AU - Ntaliarda, G. AU - Skiadas, G. AU - Lamort, A.-S. AU - Vreka, M. AU - Marazioti, A. AU - Lilis, I. AU - Bouloukou, E. AU - Giotopoulou, G.A. AU - Pepe, M. AU - Weiss, S.A. AU - Petrera, A. AU - Hauck, S.M. AU - Koch, I.* AU - Lindner, M.* AU - Hatz, R.A.* AU - Behr, J.* AU - Arendt, K.A.M. AU - Giopanou, I. AU - Brunn, D.* AU - Savai, R.* AU - Jenne, D. AU - de Château, M.* AU - Yull, F.E.* AU - Blackwell, T.S.* AU - Stathopoulos, G.T. C1 - 67652 C2 - 53959 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Non-oncogene addiction of KRAS-mutant cancers to IL-1β via versican and mononuclear IKKβ. JO - Cancers VL - 15 IS - 6 PB - Mdpi PY - 2023 SN - 2072-6694 ER - TY - JOUR AB - Neural-network-based outcome predictions may enable further treatment personalization of patients with head and neck cancer. The development of neural networks can prove challenging when a limited number of cases is available. Therefore, we investigated whether multitask learning strategies, implemented through the simultaneous optimization of two distinct outcome objectives (multi-outcome) and combined with a tumor segmentation task, can lead to improved performance of convolutional neural networks (CNNs) and vision transformers (ViTs). Model training was conducted on two distinct multicenter datasets for the endpoints loco-regional control (LRC) and progression-free survival (PFS), respectively. The first dataset consisted of pre-treatment computed tomography (CT) imaging for 290 patients and the second dataset contained combined positron emission tomography (PET)/CT data of 224 patients. Discriminative performance was assessed by the concordance index (C-index). Risk stratification was evaluated using log-rank tests. Across both datasets, CNN and ViT model ensembles achieved similar results. Multitask approaches showed favorable performance in most investigations. Multi-outcome CNN models trained with segmentation loss were identified as the optimal strategy across cohorts. On the PET/CT dataset, an ensemble of multi-outcome CNNs trained with segmentation loss achieved the best discrimination (C-index: 0.29, 95% confidence interval (CI): 0.22-0.36) and successfully stratified patients into groups with low and high risk of disease progression (p=0.003). On the CT dataset, ensembles of multi-outcome CNNs and of single-outcome ViTs trained with segmentation loss performed best (C-index: 0.26 and 0.26, CI: 0.18-0.34 and 0.18-0.35, respectively), both with significant risk stratification for LRC in independent validation (p=0.002 and p=0.011). Further validation of the developed multitask-learning models is planned based on a prospective validation study, which has recently completed recruitment. AU - Starke, S.* AU - Zwanenburg, A.* AU - Leger, K.* AU - Lohaus, F.* AU - Linge, A.* AU - Kalinauskaite, G.* AU - Tinhofer, I.* AU - Guberina, N.* AU - Guberina, M.* AU - Balermpas, P.* AU - Grün, J.V.* AU - Ganswindt, U. AU - Belka, C. AU - Peeken, J.C. AU - Combs, S.E. AU - Boeke, S.* AU - Zips, D.* AU - Richter, C.* AU - Troost, E.G.C.* AU - Krause, M.* AU - Baumann, M.* AU - Löck, S.* C1 - 68654 C2 - 54858 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Multitask learning with convolutional neural networks and vision transformers for outcome prediction of head and neck cancer patients JO - Cancers VL - 15 IS - 19 PB - Mdpi PY - 2023 SN - 2072-6694 ER - TY - JOUR AB - Glioblastoma multiforme (GBM) is the most common malignant primary brain tumor in adults. Despite modern, multimodal therapeutic options of surgery, chemotherapy, tumor-treating fields (TTF), and radiotherapy, the 5-year survival is below 10%. In order to develop new therapies, better preclinical models are needed that mimic the complexity of a tumor. In this work, we established a novel three-dimensional (3D) model for patient-derived GBM cell lines. To analyze the volume and growth pattern of primary GBM cells in 3D culture, a CoSeedisTM culture system was used, and radiation sensitivity in comparison to conventional 2D colony formation assay (CFA) was analyzed. Both culture systems revealed a dose-dependent reduction in survival, but the high variance in colony size and shape prevented reliable evaluation of the 2D cultures. In contrast, the size of 3D spheroids could be measured accurately. Immunostaining of spheroids grown in the 3D culture system showed an increase in the DNA double-strand-break marker γH2AX one hour after irradiation. After 24 h, a decrease in DNA damage was observed, indicating active repair mechanisms. In summary, this new translational 3D model may better reflect the tumor complexity and be useful for analyzing the growth, radiosensitivity, and DNA repair of patient-derived GBM cells. AU - Strand, Z.* AU - Schrickel, F.* AU - Dobiasch, S. AU - Thomsen, A.R.* AU - Steiger, K.* AU - Gempt, J.* AU - Meyer, B.* AU - Combs, S.E. AU - Schilling, D. C1 - 67948 C2 - 54426 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Establishment of a 3D model to characterize the radioresponse of patient-derived glioblastoma cells. JO - Cancers VL - 15 IS - 16 PB - Mdpi PY - 2023 SN - 2072-6694 ER - TY - JOUR AB - (1) Purpose: To assess the safety and effectivity of stereotactic body radiotherapy (SBRT) on spinal metastases utilizing a simultaneous integrated boost (SIB) concept in oligometastatic cancer patients. (2) Methods: 62 consecutive patients with 71 spinal metastases received SIB-SBRT between 01/2013 and 09/2022 at our institution. We retrospectively analyzed toxicity, local tumor control (LC), and progression-free (PFS) and overall survival (OS) following SIB-SBRT and assessed possible influencing factors (Kaplan-Meier estimator, log-rank test and Cox proportional-hazards model). (3) Results: SIB-SBRT was delivered in five fractions, mostly with 25/40 Gy (n = 43; 60.56%) and 25/35 Gy (n = 19, 26.76%). Estimated rates of freedom from VCF were 96.1/90.4% at one/two years. VCF development was significantly associated with osteoporosis (p < 0.001). No ≥ grade III acute and one grade III late toxicity (VCF) were observed. Estimated LC rates at one/two years were 98.6/96.4%, and histology was significantly associated with local treatment failure (p = 0.039). Median PFS/OS was 10 months (95% CI 6.01-13.99)/not reached. Development of metastases ≥ one year after initial diagnosis and Karnofsky Performance Score ≥ 90% were predictors for superior PFS (p = 0.038) and OS (p = 0.012), respectively. (4) Conclusion: Spinal SIB-SBRT yields low toxicity and excellent LC. It may be utilized in selected oligometastatic patients to improve prognosis. To the best of our knowledge, we provide the first clinical data on the toxicity and effectivity of SIB-SBRT in spinal metastases in a larger patient cohort. AU - Waltenberger, M.* AU - Strick, C.* AU - Vogel, M.M.E.* AU - Diehl, C. AU - Combs, S.E. C1 - 69040 C2 - 53822 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - SBRT of spinal metastases using a simultaneous integrated boost concept in oligometastatic cancer patients is safe and effective. JO - Cancers VL - 15 IS - 24 PB - Mdpi PY - 2023 SN - 2072-6694 ER - TY - JOUR AB - A strong association between the proportion of indigenous South American Mapuche ancestry and the risk of gallbladder cancer (GBC) has been reported in observational studies. Chileans show the highest incidence of GBC worldwide, and the Mapuche are the largest indigenous people in Chile. We set out to assess the confounding-free effect of the individual proportion of Mapuche ancestry on GBC risk and to investigate the mediating effects of gallstone disease and body mass index (BMI) on this association. Genetic markers of Mapuche ancestry were selected based on the informativeness for assignment measure, and then used as instrumental variables in two-sample Mendelian randomization analyses and complementary sensitivity analyses. Results suggested a putatively causal effect of Mapuche ancestry on GBC risk (inverse variance-weighted (IVW) risk increase of 0.8% per 1% increase in Mapuche ancestry proportion, 95% CI 0.4% to 1.2%, p = 6.7 × 10-5) and also on gallstone disease (3.6% IVW risk increase, 95% CI 3.1% to 4.0%), pointing to a mediating effect of gallstones on the association between Mapuche ancestry and GBC. In contrast, the proportion of Mapuche ancestry showed a negative effect on BMI (IVW estimate -0.006 kg/m2, 95% CI -0.009 to -0.003). The results presented here may have significant implications for GBC prevention and are important for future admixture mapping studies. Given that the association between the individual proportion of Mapuche ancestry and GBC risk previously noted in observational studies appears to be free of confounding, primary and secondary prevention strategies that consider genetic ancestry could be particularly efficient. AU - Zollner, L.* AU - Boekstegers, F.* AU - Barahona Ponce, C.* AU - Scherer, D.* AU - Marcelain, K.* AU - Garate-Calderon, V.* AU - Waldenberger, M. AU - Morales, E.* AU - Rojas, A.* AU - Muñoz, C.* AU - Retamales, J.* AU - de Toro, G.* AU - Kortmann, A.V.* AU - Barajas, O.* AU - Rivera, M.T.* AU - Cortes, A.* AU - Loader, D.* AU - Saavedra, J.* AU - Gutierrez, L.* AU - Ortega, A.* AU - Bertran, M.E.* AU - Bartolotti, L.* AU - Gabler, F.* AU - Campos, M.* AU - Alvarado, J.* AU - Moisán, F.* AU - Spencer, L.* AU - Nervi, B.* AU - Carvajal, D.* AU - Losada, H.* AU - Almau, M.* AU - Fernandez, P.* AU - Olloquequi, J.* AU - Carter, A.R.* AU - Miquel Poblete, J.F.* AU - Bustos, B.I.* AU - Fuentes Guajardo, M.* AU - Gonzalez-Jose, R.* AU - Bortolini, M.C.* AU - Acuña-Alonzo, V.* AU - Gallo, C.* AU - Ruiz Linares, A.* AU - Rothhammer, F.* AU - Lorenzo Bermejo, J.* C1 - 67950 C2 - 54428 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Gallbladder cancer risk and indigenous South American Mapuche ancestry : Instrumental variable analysis using ancestry-informative markers. JO - Cancers VL - 15 IS - 16 PB - Mdpi PY - 2023 SN - 2072-6694 ER - TY - JOUR AB - Long noncoding RNAs (lncRNAs) play key roles in cell processes and are good candi-dates for cancer risk prediction. Few studies have investigated the association between individual genotypes and lncRNA expression. Here we integrate three separate datasets with information on lncRNA expression only, both lncRNA expression and genotype, and genotype information only to identify circulating lncRNAs associated with the risk of gallbladder cancer (GBC) using robust linear and logistic regression techniques. In the first dataset, we preselect lncRNAs based on expression changes along the sequence “gallstones → dysplasia → GBC”. In the second dataset, we validate associations between genetic variants and serum expression levels of the preselected lncR-NAs (cis-lncRNA-eQTLs) and build lncRNA expression prediction models. In the third dataset, we predict serum lncRNA expression based on individual genotypes and assess the association between genotype-based expression and GBC risk. AC084082.3 and LINC00662 showed increasing expression levels (p-value = 0.009), while C22orf34 expression decreased in the sequence from gallstones to GBC (p-value = 0.04). We identified and validated two cis-LINC00662-eQTLs (r2 = 0.26) and three cis-C22orf34-eQTLs (r2 = 0.24). Only LINC00662 showed a genotyped-based serum expression associated with GBC risk (OR = 1.25 per log2 expression unit, 95% CI 1.04–1.52, p-value = 0.02). Our results suggest that preselection of lncRNAs based on tissue samples and exploitation of cis-lncRNA-eQTLs may facilitate the identification of circulating noncoding RNAs linked to cancer risk. AU - Blandino, A.* AU - Scherer, D.* AU - Rounge, T.B.* AU - Umu, S.U.* AU - Boekstegers, F.* AU - Barahona Ponce, C.* AU - Marcelain, K.* AU - Garate-Calderon, V.* AU - Waldenberger, M. AU - Morales, E.* AU - Rojas, A.* AU - Muñoz, C.* AU - Retamales, J.* AU - de Toro, G.* AU - Barajas, O.* AU - Rivera, M.T.* AU - Cortes, A.* AU - Loader, D.* AU - Saavedra, J.M.* AU - Gutiérrez, L.* AU - Ortega, A.* AU - Bertrán, M.E.* AU - Gabler, F.* AU - Campos, M.* AU - Alvarado, J.* AU - Moisán, F.* AU - Spencer, L.* AU - Nervi, B.* AU - Carvajal-Hausdorf, D.E.* AU - Losada, H.* AU - Almau, M.* AU - Fernandez, P.* AU - Gallegos, I.* AU - Olloquequi, J.* AU - Fuentes-Guajardo, M.* AU - Gonzalez-Jose, R.* AU - Bortolini, M.C.* AU - Gallo, C.* AU - Ruiz Linares, A.* AU - Rothhammer, F.* AU - Bermejo, J.L.* C1 - 64164 C2 - 51998 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Identification of circulating lncRNAs associated with gallbladder cancer risk by tissue-based preselection, cis-eQTL validation, and analysis of association with genotype-based expression. JO - Cancers VL - 14 IS - 3 PB - Mdpi PY - 2022 SN - 2072-6694 ER - TY - JOUR AB - Rare variants in at least 10 genes, including BRCA1, BRCA2, PALB2, ATM, and CHEK2, are associated with increased risk of breast cancer; however, these variants, in combination with common variants identified through genome-wide association studies, explain only a fraction of the familial aggregation of the disease. To identify further susceptibility genes, we performed a two-stage whole-exome sequencing study. In the discovery stage, samples from 1528 breast cancer cases enriched for breast cancer susceptibility and 3733 geographically matched unaffected controls were sequenced. Using five different filtering and gene prioritization strategies, 198 genes were selected for further validation. These genes, and a panel of 32 known or suspected breast cancer susceptibility genes, were assessed in a validation set of 6211 cases and 6019 controls for their association with risk of breast cancer overall, and by estrogen receptor (ER) disease subtypes, using gene burden tests applied to loss-of-function and rare missense variants. Twenty genes showed nominal evidence of association (p-value < 0.05) with either overall or subtype-specific breast cancer. Our study had the statistical power to detect susceptibility genes with effect sizes similar to ATM, CHEK2, and PALB2, however, it was underpowered to identify genes in which susceptibility variants are rarer or confer smaller effect sizes. Larger sample sizes would be required in order to identify such genes. AU - Dumont, M.* AU - Weber-Lassalle, N.* AU - Joly-Beauparlant, C.* AU - Ernst, C.* AU - Droit, A.* AU - Feng, B.* AU - Dubois, S.* AU - Collin-Deschesnes, A.* AU - Soucy, P.* AU - Vallee, M.* AU - Fournier, F.* AU - Lemacon, A.* AU - Adank, M.A.* AU - Allen, J.* AU - Altmueller, J.* AU - Arnold, N.* AU - Ausems, M.G.E.M.* AU - Berutti, R.* AU - Bolla, M.K.* AU - Bull, S.* AU - Carvalho, S.* AU - Cornelissen, S.* AU - Dufault, M.R.* AU - Dunning, A.M.* AU - Engel, C.* AU - Gehrig, A.* AU - Geurts-Giele, W.R.R.* AU - Gieger, C. AU - Green, J.* AU - Hackmann, K.* AU - Helmy, M.* AU - Hentschel, J.* AU - Hogervorst, F.B.L.* AU - Hollestelle, A.* AU - Hooning, M.J.* AU - Horvath, J.* AU - Ikram, M.A.A.* AU - Kaulfuss, S.* AU - Keeman, R.* AU - Kuang, D.* AU - Luccarini, C.* AU - Maier, W.* AU - Martens, J.W.M.* AU - Niederacher, D.* AU - Nürnberg, P.* AU - Ott, C.* AU - Peters, A. AU - Pharoah, P.D.P.* AU - Ramirez, A.* AU - Ramser, J.* AU - Riedel-Heller, S.* AU - Schmidt, G.* AU - Shah, M.* AU - Scherer, M.* AU - Stäbler, A.* AU - Strom, T.M.* AU - Sutter, C.* AU - Thiele, H.* AU - van Asperen, C.J.* AU - van der Kolk, L.* AU - van der Luijt, R.B.* AU - Volk, A.E.* AU - Wagner, M.* AU - Waisfisz, Q.* AU - Wang, Q.* AU - Wang-Gohrke, S.* AU - Weber, B.H.F.* AU - Devilee, P.* AU - Tavtigian, S.* AU - Bader, G.D.* AU - Meindl, A.* AU - Goldgar, D.E.* AU - Andrulis, I.L.* AU - Schmutzler, R.K.* AU - Easton, D.F.* AU - Schmidt, M.K.* AU - Hahnen, E.* AU - Simard, J.* C1 - 65838 C2 - 52534 TI - Uncovering the contribution of moderate-penetrance susceptibility genes to breast cancer by whole-exome sequencing and targeted enrichment sequencing of candidate genes in women of European ancestry. JO - Cancers VL - 14 IS - 14 PY - 2022 SN - 2072-6694 ER - TY - JOUR AB - High mass resolution matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) is a suitable method for biomarker detection for several tumor entities. Renal cell carcinoma (RCC) is the seventh most common cancer type and accounts for more than 80% of all renal tumors. Prognostic biomarkers for RCC are still missing. Therefore, we analyzed a large, multicenter cohort including the three most common RCC subtypes (clear cell RCC (ccRCC), papillary RCC (pRCC) and chromophobe RCC (chRCC)) by MALDI for prognostic biomarker detection. MALDI-Fourier-transform ion cyclotron resonance (FT-ICR)-MSI analysis was performed for renal carcinoma tissue sections from 782 patients. SPACiAL pipeline was integrated for automated co-registration of histological and molecular features. Kaplan–Meier analyses with overall survival as endpoint were executed to determine the metabolic features associated with clinical outcome. We detected several pathways and metabolites with prognostic power for RCC in general and also for different RCC subtypes. AU - Erlmeier, F.* AU - Sun, N. AU - Shen, J. AU - Feuchtinger, A. AU - Buck, A. AU - Prade, V.M. AU - Kunzke, T. AU - Schraml, P.* AU - Moch, H.* AU - Autenrieth, M.* AU - Weichert, W.* AU - Hartmann, A.* AU - Walch, A.K. C1 - 64744 C2 - 52431 TI - MALDI mass spectrometry imaging—prognostic pathways and metabolites for renal cell carcinomas. JO - Cancers VL - 14 IS - 7 PY - 2022 SN - 2072-6694 ER - TY - JOUR AB - In the past, different bacterial species have been tested for cancer therapy in preclinical and clinical studies. The success of bacterial cancer therapy is mainly dependent on the ability of the utilized bacteria to overcome the host immune defense system to colonize the tumors and to initiate tumor-specific immunity. In recent years, several groups have demonstrated that the gut microbiome plays an important role of modulation of the host immune response and has an impact on therapeutic responses in murine models and in cohorts of human cancer patients. Here we analyzed the impact of the gut microbiome on tumor colonization and tumor therapy by the Escherichia coli Nissle 1917 (EcN) strain. This EcN strain is a promising cancer therapy candidate with probiotic properties. In our study, we observed significantly better tumor colonization by EcN after antibiotic-induced temporal depletion of the gut microbiome and after two intranasal applications of the EcN derivate (EcN/pMUT-gfp Knr) in 4T1 tumor-bearing syngeneic BALB/c mice. In addition, we demonstrated significant reduction in tumor growth and extended survival of the EcN-treated mice in contrast to phosphate-buffered saline (PBS)-treated tumor-bearing control animals. Multispectral imaging of immune cells revealed that depletion of the gut microbiome led to significantly lower infiltration of cytotoxic and helper T cells (CD4 and CD8 cells) in PBS tumors of mice pretreated with antibiotics in comparison with antibiotic untreated PBS—or EcN treated mice. These findings may help in the future advancement of cancer treatment strategies using E. coli Nissle 1917. AU - Gentschev, I.* AU - Petrov, I.* AU - Ye, M.* AU - Kafuri Cifuentes, L.* AU - Toews, R.* AU - Cecil, A. AU - Oelschaeger, T.A.* AU - Szalay, A.A.* C1 - 67111 C2 - 53443 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Tumor colonization and therapy by Escherichia coli nissle 1917 strain in syngeneic tumor-bearing mice is strongly affected by the gut microbiome. JO - Cancers VL - 14 IS - 24 PB - Mdpi PY - 2022 SN - 2072-6694 ER - TY - JOUR AB - Background: Cancer is primarily a disease of high age in humans, yet most mouse studies on cancer cachexia are conducted using young adolescent mice. Given that metabolism and muscle function change with age, we hypothesized that aging may affect cachexia progression in mouse models. Methods: We compare tumor and cachexia development in young and old mice of three different strains (C57BL/6J, C57BL/6N, BALB/c) and with two different tumor cell lines (Lewis Lung Cancer, Colon26). Tumor size, body and organ weights, fiber cross-sectional area, circulating cachexia biomarkers, and molecular markers of muscle atrophy and adipose tissue wasting are shown. We correlate inflammatory markers and body weight dependent on age in patients with cancer. Results: We note fundamental differences between mouse strains. Aging aggravates weight loss in LLC-injected C57BL/6J mice, drives it in C57BL/6N mice, and does not influence weight loss in C26-injected BALB/c mice. Glucose tolerance is unchanged in cachectic young and old mice. The stress marker GDF15 is elevated in cachectic BALB/c mice independent of age and increased in old C57BL/6N and J mice. Inflammatory markers correlate significantly with weight loss only in young mice and patients. Conclusions: Aging affects cachexia development and progression in mice in a strain-dependent manner and influences the inflammatory profile in both mice and patients. Age is an important factor to consider for future cachexia studies. AU - Geppert, J. AU - Walth-Hummel, A.A. AU - Terron Exposito, R. AU - Kaltenecker, D. AU - Morigny, P. AU - Machado, J. AU - Becker, M. AU - Simoes Fernandez, E. AU - Lima, J.D.C.C.* AU - Daniel, C. AU - Diaz, M.B. AU - Herzig, S. AU - Seelaender, M.* AU - Rohm, M. C1 - 63925 C2 - 51721 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Aging aggravates cachexia in tumor-bearing mice. JO - Cancers VL - 14 IS - 1 PB - Mdpi PY - 2022 SN - 2072-6694 ER - TY - JOUR AB - Pancreatic neuroendocrine neoplasms (PanNENs) are the second most common malignancy of the pancreas. Surgery remains the only curative treatment for localized disease. For patients with inoperable advanced or metastatic disease, few targeted therapies are available, but their efficacy is unpredictable and variable. Exploiting prior knowledge on pathogenetic processes involved in PanNEN tumorigenesis, we tested buparlisib (PI3K inhibitor) and ribociclib (CDK4/6 inhibitor), as single agents or in combination, in different preclinical models. First, we used cell lines representative of well-differentiated (INS-1E, NT-3) and poorly differentiated (BON-1) PanNENs. The combination of buparlisib with ribociclib reduced the proliferation of 2D and 3D spheroid cultures more potently than the individual drugs. Buparlisib, but not ribociclib, induced apoptosis. The anti-proliferative activity of the drugs correlated with downstream target inhibition at mRNA and protein levels. We then tested the drugs on primary islet microtissues from a genetic PanNET animal model (Men1-defective mice) and from wild-type mice: the drug combination was effective against the former without altering islet cell physiology. Finally, we treated PanNET patient-derived islet-like 3D tumoroids: the combination of buparlisib with ribociclib was effective in three out of four samples. Combined targeting of PI3K and CDK4/6 is a promising strategy for PanNENs spanning various molecular and histo-pathological features. AU - Gulde, S. AU - Foscarini, A. AU - April-Monn, S.L.* AU - Genio, E. AU - Marangelo, A. AU - Satam, S. AU - Helbling, D.* AU - Falconi, M.* AU - Toledo, R.A.* AU - Schrader, J.* AU - Perren, A.* AU - Marinoni, I.* AU - Pellegata, N.S. C1 - 66831 C2 - 53305 TI - Combined targeting of pathogenetic mechanisms in pancreatic neuroendocrine tumors elicits synergistic antitumor effects. JO - Cancers VL - 14 IS - 22 PY - 2022 SN - 2072-6694 ER - TY - JOUR AB - Human papillomavirus (HPV)-driven head and neck squamous cell carcinomas (HNSCC) generally have a more favourable prognosis. We hypothesized that HPV-associated HNSCC may be identified by an miRNA-signature according to their specific molecular pathogenesis, and be characterized by a unique transcriptome compared to HPV-negative HNSCC. We performed miRNA expression profiling of two p16/HPV DNA characterized HNSCC cohorts of patients treated by adjuvant radio(chemo)therapy (multicentre DKTK-ROG n = 128, single-centre LMU-KKG n = 101). A linear model predicting HPV status built in DKTK-ROG using lasso-regression was tested in LMU-KKG. LMU-KKG tumours (n = 30) were transcriptome profiled for differential gene expression and miRNA-integration. A 24-miRNA signature predicted HPV-status with 94.53% accuracy (AUC: 0.99) in DKTK-ROG, and 86.14% (AUC: 0.86) in LMU-KKG. The prognostic values of 24-miRNA- and p16/HPV DNA status were comparable. Combining p16/HPV DNA and 24-miRNA status allowed patient sub-stratification and identification of an HPV-associated patient subgroup with impaired overall survival. HPV-positive tumours showed downregulated MAPK, Estrogen, EGFR, TGFbeta, WNT signaling activity. miRNA-mRNA integration revealed HPV-specific signaling pathway regulation, including PD-L1 expression/PD-1 checkpoint pathway in cancer in HPV-associated HNSCC. Integration of clinically established p16/HPV DNA with 24-miRNA signature status improved clinically relevant risk stratification, which might be considered for future clinical decision-making with respect to treatment de-escalation in HPV-associated HNSCC. AU - Hess J. AU - Unger, K. AU - Maihoefer, C. AU - Schuettrumpf, L. AU - Weber, P. AU - Marschner, S. AU - Wintergerst, L. AU - Pflugradt, U. AU - Baumeister, P. AU - Walch, A.K. AU - Woischke, C.* AU - Kirchner, T.* AU - Werner, M.* AU - Soerensen, K.* AU - Baumann, M.* AU - Tinhofer, I.* AU - Combs, S.E. AU - Debus, J.* AU - Schaefer, H.* AU - Krause, M.* AU - Linge, A.* AU - von der Gruen, J.* AU - Stuschke, M.* AU - Zips, D.* AU - Canis, M. AU - Lauber, K. AU - Ganswindt, U. AU - Henke, M.* AU - Zitzelsberger, H. AU - Belka, C. C1 - 65930 C2 - 52638 TI - Integration of p16/HPV DNA status with a 24-miRNA-defined molecular phenotype improves clinically relevant stratification of head and neck cancer patients. JO - Cancers VL - 14 IS - 15 PY - 2022 SN - 2072-6694 ER - TY - JOUR AB - The current study aims to assess the suitability of setup errors during the first three treatment fractions to determine cone-beam computed tomography (CBCT) frequency in adjuvant breast radiotherapy. For this, 45 breast cancer patients receiving non-hypofractionated radiotherapy after lumpectomy, including a simultaneous integrated boost (SIB) to the tumor bed and daily CBCT imaging, were retrospectively selected. In a first step, mean and maximum setup errors on treatment days 1-3 were correlated with the mean setup errors during subsequent treatment days. In a second step, dose distribution was estimated using a dose accumulation workflow based on deformable image registration, and setup errors on treatment days 1-3 were correlated with dose deviations in the clinical target volumes (CTV) and organs at risk (OAR). No significant correlation was found between mean and maximum setup errors on treatment days 1-3 and mean setup errors during subsequent treatment days. In addition, mean and maximum setup errors on treatment days 1-3 correlated poorly with dose coverage of the CTVs and dose to the OARs. Thus, CBCT frequency in adjuvant breast radiotherapy should not be determined solely based on the magnitude of setup errors during the first three treatment fractions. AU - Junker, Y.* AU - Düsberg, M.* AU - Asadpour, R.* AU - Klusen, S.* AU - Münch, S.* AU - Bernhardt, D.* AU - Combs, S.E. AU - Borm, K.J.* C1 - 66159 C2 - 53097 TI - As easy as 1, 2, 3? How to determine CBCT frequency in adjuvant breast radiotherapy. JO - Cancers VL - 14 IS - 17 PY - 2022 SN - 2072-6694 ER - TY - JOUR AB - Microbeam radiotherapy (MRT) is a novel, still preclinical dose delivery technique. MRT has shown reduced normal tissue effects at equal tumor control rates compared to conventional radiotherapy. Treatment planning studies are required to permit clinical application. The aim of this study was to establish a dose comparison between MRT and conventional radiotherapy and to identify suitable clinical scenarios for future applications of MRT. We simulated MRT treatment scenarios for clinical patient data using an inhouse developed planning algorithm based on a hybrid Monte Carlo dose calculation and implemented the concept of equivalent uniform dose (EUD) for MRT dose evaluation. The investigated clinical scenarios comprised fractionated radiotherapy of a glioblastoma resection cavity, a lung stereotactic body radiotherapy (SBRT), palliative bone metastasis irradiation, brain metastasis radiosurgery and hypofractionated breast cancer radiotherapy. Clinically acceptable treatment plans were achieved for most analyzed parameters. Lung SBRT seemed the most challenging treatment scenario. Major limitations comprised treatment plan optimization and dose calculation considering the tissue microstructure. This study presents an im-portant step of the development towards clinical MRT. For clinical treatment scenarios using a so-phisticated dose comparison concept based on EUD and EQD2, we demonstrated the capability of MRT to achieve clinically acceptable dose distributions. AU - Kraus, K.M. AU - Winter, J. AU - Zhang, Y. AU - Ahmed, M. AU - Combs, S.E. AU - Wilkens, J.J.* AU - Bartzsch, S. C1 - 64194 C2 - 52099 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Treatment planning study for microbeam radiotherapy using clinical patient data. JO - Cancers VL - 14 IS - 3 PB - Mdpi PY - 2022 SN - 2072-6694 ER - TY - JOUR AB - Thoracic stereotactic body radiation therapy (SBRT) is extensively used in combination with immune checkpoint blockade (ICB). While current evidence suggests that the occurrence of pneumonitis as a side effect of both treatments is not enhanced for the combination, the dose-volume correlation remains unclear. We investigate dose-volume-effect correlations for pneumonitis after combined SBRT + ICB. We analyzed patient clinical characteristics and dosimetric data for 42 data sets for thoracic SBRT with ICB treatment (13) and without (29). Dose volumes were converted into 2 Gy equivalent doses (EQD2), allowing for dosimetric comparison of different fractionation regimes. Pneumonitis volumes were delineated and corresponding DVHs were analyzed. We noticed a shift towards lower doses for combined SBRT + ICB treatment, supported by a trend of smaller areas under the curve (AUC) for SBRT+ ICB (median AUC 1337.37 vs. 5799.10, p = 0.317). We present a DVH-based dose-volume-effect correlation method and observed large pneumonitis volumes, even with bilateral extent in the SBRT + ICB group. We conclude that further studies using this method with enhanced statistical power are needed to clarify whether adjustments of the radiation dose constraints are required to better estimate risks of pneumonitis after the combination of SBRT and ICB. AU - Kraus, K.M. AU - Bauer, C.* AU - Feuerecker, B.* AU - Fischer, J.C.* AU - Borm, K.J.* AU - Bernhardt, D.* AU - Combs, S.E. C1 - 65532 C2 - 52722 TI - Pneumonitis after stereotactic thoracic radioimmunotherapy with checkpoint inhibitors: Exploration of the dose-volume-effect correlation. JO - Cancers VL - 14 IS - 12 PY - 2022 SN - 2072-6694 ER - TY - JOUR AB - BACKGROUND: To evaluate differences in local tumor control (LC), symptoms and quality of life (QOL) of 261 patients with VS after stereotactic radiosurgery/hypofractionated stereotactic radiotherapy (SRS/HFSRT) vs. fractionated radiotherapy (FRT) vs. fractionated proton therapy (FPT) were studied. METHODS: For SRS/HFSRT (n = 149), the median fraction dose applied was 12 Gy. For FRT (n = 87) and FPT (n = 25), the median cumulative doses applied were 57.6 Gy and 54 Gy (RBE), respectively. FRT and FPT used single median doses of 1.8 Gy/Gy (RBE). Median follow-up was 38 months. We investigated dosimetry for organs at risk and analyzed toxicity and QOL by sending out a questionnaire. RESULTS: LC was 99.5% at 12 months after RT with no statistical difference between treatment groups (p = 0.19). LC was significantly lower in NF2 patients (p = 0.004) and in patients with higher tumor extension grade (p = 0.039). The hearing preservation rate was 97% at 12 months after RT with no statistical difference between treatment groups (p = 0.31). Facial and trigeminal nerve affection after RT occurred as mild symptoms with highest toxicity rate in FPT patients. CONCLUSION: SRS/HFSRT, FRT and FPT for VS show similar overall clinical and functional outcomes. Cranial nerve impairment rates vary, potentially due to selection bias with larger VS in the FRT and FPT group. AU - Küchler, M.* AU - El Shafie, R.A.* AU - Herfarth, K.* AU - König, L.* AU - Lang, K.* AU - Hörner-Rieber, J.* AU - Plinkert, P.K.* AU - Wick, W.* AU - Sahm, F.* AU - Sprengel, S.D.* AU - Bernhardt, D. C1 - 64877 C2 - 52575 TI - Outcome after radiotherapy for vestibular schwannomas (VS)-differences in tumor control, symptoms and quality of life after radiotherapy with photon versus proton therapy. JO - Cancers VL - 14 IS - 8 PY - 2022 SN - 2072-6694 ER - TY - JOUR AB - Acute myeloid leukemia (AML) is the most common acute leukemia in adults. Patients with AML harboring a constitutively active internal tandem duplication mutation (ITDMUT ) in the FMS-like kinase tyrosine kinase (FLT3) receptor generally have a poor prognosis. Several tyrosine kinase/FLT3 inhibitors have been developed and tested clinically, but very few (midostaurin and gilteritinib) have thus far been FDA/EMA-approved for patients with newly diagnosed or relapse/refractory FLT3-ITDMUT AML. Disappointingly, clinical responses are commonly partial or not durable, highlighting the need for new molecules targeting FLT3-ITDMUT AML. Here, we tested EC-70124, a hybrid indolocarbazole analog from the same chemical space as midostaurin with a potent and selective inhibitory effect on FLT3. In vitro, EC-70124 exerted a robust and specific antileukemia activity against FLT3-ITDMUT AML primary cells and cell lines with respect to cytotoxicity, CFU capacity, apoptosis and cell cycle while sparing healthy hematopoietic (stem/progenitor) cells. We also analyzed its efficacy in vivo as monotherapy using two different xenograft models: an aggressive and systemic model based on MOLM-13 cells and a patient-derived xenograft model. Orally disposable EC-70124 exerted a potent inhibitory effect on the growth of FLT3-ITDMUT AML cells, delaying disease progression and debulking the leukemia. Collectively, our findings show that EC-70124 is a promising and safe agent for the treatment of AML with FLT3-ITDMUT. AU - Lopez-Millan, B.* AU - Costales, P.* AU - Gutiérrez-Agüera, F.* AU - de la Guardia, R.D.* AU - Roca-Ho, H.* AU - Vinyoles, M.* AU - Rubio-Gayarre, A.* AU - Safi, R.* AU - Castaño, J.* AU - Romecín, P.A.* AU - Ramírez-Orellana, M.* AU - Anguita, E.* AU - Jeremias, I. AU - Zamora, L.* AU - Rodríguez-Manzaneque, J.C.* AU - Bueno, C.* AU - Morís, F.* AU - Menendez, P.* C1 - 64657 C2 - 52397 TI - The multi-kinase inhibitor EC-70124 is a promising candidate for the treatment of FLT3-ITD-positive acute myeloid leukemia. JO - Cancers VL - 14 IS - 6 PY - 2022 SN - 2072-6694 ER - TY - JOUR AB - The aim of the study was to develop a new therapeutic strategy to target cancer stem cells (CSCs) in clear cell renal cell carcinoma (ccRCC) and to identify typical CSC markers to improve therapy effectiveness. It was found that the corrected-mRNA expression-based stemness index was upregulated in kidney renal clear cell carcinoma (KIRC) tissues compared to non-tumor tissue and increased with higher tumor stage and grade. EZH2 was identified as a CSC marker and prognosis factor for KIRC patients. The expression of EZH2 was associated with several activated tumor-infiltrating immune cells. High expression of EZH2 was enriched in immune-related pathways, low expression was related to several metabolic pathways. Epigallocatechin-3-gallate (EGCG) was identified as the most potent suppressor of EZH2, was able to inhibit viability, migration, and invasion, and to increase the apoptosis rate of ccRCC CSCs. KIF11, VEGF, and MMP2 were identified as predictive EGCG target genes, suggesting a potential mechanism of how EZH2 might regulate invasiveness and migration. The percentages of FoxP3+ Treg cells in the peripheral blood mononuclear cells of ccRCC patients decreased significantly when cultured with spheres pretreated with EGCG plus sunitinib compared to spheres without treatment. Our findings provide new insights into the treatment options of ccRCC based on targeting CSCs. AU - Lyu, C.* AU - Wang, L.* AU - Stadlbauer, B.* AU - Nößner, E. AU - Buchner, A.* AU - Pohla, H.* C1 - 66158 C2 - 53096 TI - Identification of EZH2 as cancer stem cell marker in clear cell renal cell carcinoma and the anti-tumor effect of epigallocatechin-3-gallate (EGCG). JO - Cancers VL - 14 IS - 17 PY - 2022 SN - 2072-6694 ER - TY - JOUR AB - (1) Background: Patients with locally advanced head and neck squamous cell carcinoma (HNSCC) who are biologically at high risk for the development of loco-regional recurrences after postoperative radiotherapy (PORT) but at intermediate risk according to clinical risk factors may benefit from additional concurrent chemotherapy. In this matched-pair study, we aimed to identify a corresponding predictive gene signature. (2) Methods: Gene expression analysis was performed on a multicenter retrospective cohort of 221 patients that were treated with postoperative radiochemotherapy (PORT-C) and 283 patients who were treated with PORT alone. Propensity score analysis was used to identify matched patient pairs from both cohorts. From differential gene expression analysis and Cox regression, a predictive gene signature was identified. (3) Results: 108 matched patient pairs were selected. We identified a 2-metagene signature that stratified patients into risk groups in both cohorts. The comparison of the high-risk patients between the two types of treatment showed higher loco-regional control (LRC) after treatment with PORT-C (p < 0.001), which was confirmed by a significant interaction term in Cox regression (p = 0.027), i.e., the 2-metagene signature was indicative for the type of treatment. (4) Conclusion: We have identified a novel gene signature that may be helpful to identify patients with high-risk HNSCC amongst those at intermediate clinical risk treated with PORT, who may benefit from additional concurrent chemotherapy. AU - Patil, S.* AU - Linge, A.* AU - Hiepe, H.* AU - Grosser, M.* AU - Lohaus, F.* AU - Gudziol, V.* AU - Kemper, M.* AU - Nowak, A.* AU - Haim, D.* AU - Tinhofer, I.* AU - Budach, V.* AU - Guberina, M.* AU - Stuschke, M.* AU - Balermpas, P.* AU - Grün, J.V.* AU - Schäfer, H.* AU - Grosu, A.L.* AU - Ganswindt, U.* AU - Belka, C. AU - Pigorsch, S.* AU - Combs, S.E. AU - Boeke, S.* AU - Zips, D.* AU - Jöhrens, K.* AU - Baretton, G.B.* AU - Baumann, M.* AU - Löck, S.* C1 - 65531 C2 - 52721 TI - A novel 2-metagene signature to identify high-risk HNSCC patients amongst those who are clinically at intermediate risk and are treated with PORT. JO - Cancers VL - 14 IS - 12 PY - 2022 SN - 2072-6694 ER - TY - JOUR AB - The hostile tumor microenvironment (TME) is a major challenge for the treatment of solid tumors with T-cell receptor (TCR)-modified T-cells (TCR-Ts), as it negatively influences T-cell efficacy, fitness, and persistence. These negative influences are caused, among others, by the inhibitory checkpoint PD-1/PD-L1 axis. The Preferentially Expressed Antigen in Melanoma (PRAME) is a highly relevant cancer/testis antigen for TCR-T immunotherapy due to broad expression in multiple solid cancer indications. A TCR with high specificity and sensitivity for PRAME was isolated from non-tolerized T-cell repertoires and introduced into T-cells alongside a chimeric PD1-41BB receptor, consisting of the natural extracellular domain of PD-1 and the intracellular signaling domain of 4-1BB, turning an inhibitory pathway into a T-cell co-stimulatory pathway. The addition of PD1-41BB to CD8+ T-cells expressing the transgenic PRAME-TCR enhanced IFN-γ secretion, improved cytotoxic capacity, and prevented exhaustion upon repetitive re-challenge with tumor cells in vitro without altering the in vitro safety profile. Furthermore, a single dose of TCR-Ts co-expressing PD1-41BB was sufficient to clear a hard-to-treat melanoma xenograft in a mouse model, whereas TCR-Ts without PD1-41BB could not eradicate the PD-L1-positive tumors. This cutting-edge strategy supports development efforts to provide more effective TCR-T immunotherapies for the treatment of solid tumors. AU - Sailer, N.* AU - Fetzer, I.* AU - Salvermoser, M.* AU - Braun, M.* AU - Brechtefeld, D.* AU - Krendl, C.* AU - Geiger, C.* AU - Mutze, K.* AU - Nößner, E. AU - Schendel, D.J.* AU - Bürdek, M.* AU - Wilde, S.* AU - Sommermeyer, D.* C1 - 64836 C2 - 52515 TI - T-cells expressing a highly potent PRAME-specific T-cell receptor in combination with a chimeric PD1-41BB co-stimulatory receptor show a favorable preclinical safety profile and strong anti-tumor reactivity. JO - Cancers VL - 14 IS - 8 PY - 2022 SN - 2072-6694 ER - TY - JOUR AB - Monoplanar microbeam irradiation (MBI) and pencilbeam irradiation (PBI) are two new concepts of high dose rate radiotherapy, combined with spatial dose fractionation at the micrometre range. In a small animal model, we have explored the concept of integrating MBI or PBI as a simultaneously integrated boost (SIB), either at the beginning or at the end of a conventional, low-dose rate schedule of 5x4 Gy broad beam (BB) whole brain radiotherapy (WBRT). MBI was administered as array of 50 µm wide, quasi-parallel microbeams. For PBI, the target was covered with an array of 50 µm × 50 µm pencilbeams. In both techniques, the centre-to-centre distance was 400 µm. To assure that the entire brain received a dose of at least 4 Gy in all irradiated animals, the peak doses were calculated based on the daily BB fraction to approximate the valley dose. The results of our study have shown that the sequence of the BB irradiation fractions and the microbeam SIB is important to limit the risk of acute adverse effects, including epileptic seizures and death. The microbeam SIB should be integrated early rather than late in the irradiation schedule. AU - Schültke, E.* AU - Jaekel, F.* AU - Bartzsch, S. AU - Bräuer-Krisch, E.* AU - Requardt, H.* AU - Laissue, J.A.* AU - Blattmann, H.* AU - Hildebrandt, G.* C1 - 66945 C2 - 53381 TI - Good Timing Matters: The spatially fractionated high dose rate boost should come first. JO - Cancers VL - 14 IS - 23 PY - 2022 SN - 2072-6694 ER - TY - JOUR AB - Recent reports have shown a link between radiation exposure and non-cancer diseases such as radiation-induced heart disease (RIHD). Radiation exposures are often inhomogeneous, and out-of-target effects have been studied in terms of cancer risk, but very few studies have been carried out for non-cancer diseases. Here, the role of miRNAs in the pathogenesis of RIHD was investigated. C57Bl/6J female mice were whole- (WBI) or partial-body-irradiated (PBI) with 2 Gy of X-rays or sham-irradiated (SI). In PBI exposure, the lower third of the mouse body was irradiated, while the upper two-thirds were shielded. From all groups, hearts were collected 15 days or 6 months post-irradiation. The MiRNome analysis at 15 days post-irradiation showed that miRNAs, belonging to the myomiR family, were highly differentially expressed in WBI and PBI mouse hearts compared with SI hearts. Raman spectral data collected 15 days and 6 months post-irradiation showed biochemical differences among SI, WBI and PBI mouse hearts. Fibrosis in WBI and PBI mouse hearts, indicated by the increased deposition of collagen and the overexpression of genes involved in myofibroblast activation, was found 6 months post-irradiation. Using an in vitro co-culture system, involving directly irradiated skeletal muscle and unirradiated ventricular cardiac human cells, we propose the role of miR-1/133a as mediators of the abscopal response, suggesting that miRNA-based strategies could be relevant for limiting tissue-dependent reactions in non-directly irradiated tissues. AU - Tanno, B.* AU - Novelli, F.* AU - Leonardi, S.* AU - Merla, C.* AU - Babini, G.* AU - Giardullo, P.* AU - Kadhim, M.* AU - Traynor, D.* AU - Medipally, D.K.R.* AU - Meade, A.D.* AU - Lyng, F.M.* AU - Tapio, S. AU - Marchetti, L.* AU - Saran, A.* AU - Pazzaglia, S.* AU - Mancuso, M.* C1 - 65883 C2 - 52565 TI - MiRNA-mediated fibrosis in the out-of-target heart following partial-body irradiation. JO - Cancers VL - 14 IS - 14 PY - 2022 SN - 2072-6694 ER - TY - JOUR AB - Malignant pleural mesothelioma (MPM) is a rare, incurable cancer of the mesothelial cells lining the lungs and the chest wall that is mainly caused by asbestos inhalation. The molecular mechanisms of mesothelial carcinogenesis are still unclear despite comprehensive studies of the mutational landscape of MPM, and the most frequently mutated genes BAP1, NF2, CDKN2A, TP53, and TSC1 cannot cause MPM in mice in a standalone fashion. Although KRAS pathway alterations were sporadically detected in older studies employing targeted sequencing, they have been largely undetected by next generation sequencing. We recently identified KRAS mutations and copy number alterations in a significant proportion of MPM patients. Here, we review and analyze multiple human datasets and the published literature to show that, in addition to KRAS, multiple other genes of the KRAS pathway are perturbed in a significant proportion of patients with MPM. AU - Trassl, L. AU - Stathopoulos, G.T. C1 - 66157 C2 - 53095 TI - KRAS pathway alterations in malignant pleural mesothelioma: An underestimated player. JO - Cancers VL - 14 IS - 17 PY - 2022 SN - 2072-6694 ER - TY - JOUR AB - Objective: The management of petroclival meningiomas (PCMs) remains notoriously difficult due to their close association with neurovascular structures and their complex anatomy, hence the surgical paradigm change from radical to functional resection in the past. With this study, we aimed to analyze surgical and functional outcomes of a modern consecutive series of patients with PCMs. Methods: We reviewed patient charts and imaging data of 64 consecutive patients from 2006 to 2018 with a PCM resected at our institution and compared surgical and functional outcomes between subgroups stratified by surgical approach. Results: Females comprised 67.2% of patients (n = 43), with a mean age of 55 years (median 56; range 21–84). Follow-up data were available for 68.8% and reached a mean of 42.3 months (range 1–129) with a median of 28.5 months. The mean tumor diameter was 37.3 mm (standard deviation (SD) 15.4; median 37.0). Infiltration of the cavernous sinus was observed in 34 cases (53.1%), and the lesions affected the brain stem in 28 cases (43.8%). Preoperative cranial nerve palsy was observed in 73.4% of cases; trigeminal neuropathy (42.2%), hearing loss (32.8%), and impairment of vision (18.8%) were the most common. A retrosigmoid approach was employed in 47 cases (78.1%), pterional in 10 (15.6%), combined petrosal in 2 (3.1%), and transnasal and subtemporal in 1 (1.6%). Fifteen cases (23.4%) were resected in a two-staged fashion. Gross total resection (GTR) was attempted in 30 (46.9%) cases without cavernous sinus infiltration and was achieved in 21 (70.0%) of these cases. Surgical complications occurred in 13 cases (20.3%), most commonly meningitis (n = 4; 6.3%). Postoperatively, 56 patients (87.5%) developed new cranial nerve palsy, of which 36 (63.6%) had improved or resolved on last follow up. Achieving GTR was not significantly associated with higher rates of surgical complications (chi-square; p = 0.288) or postoperative cranial nerve palsy (chi-square; p = 0.842). Of all cases, 20 (31.3%) underwent postoperative radiation. Tumor progression was observed in 10 patients (15.9%) after a mean 102 months (median 124). Conclusions: Surgical resection remains the mainstay of treatment for PCMs, with perioperative cranial neuropathies exhibiting favorable recovery rates. Most essentially, the preselection of patients with hallmarks of brain stem affection and cavernous sinus infiltration should dictate whether to strive for a functionally oriented strategy in favor of radical resection. AU - Wagner, A.* AU - Alraun, M.* AU - Kahlig, V.* AU - Dorier, A.S.* AU - Aftahy, A.K.* AU - Bernhardt, D.* AU - Combs, S.E. AU - Gempt, J.* AU - Shiban, E.* AU - Meyer, B.* AU - Negwer, C.* C1 - 66292 C2 - 53129 TI - Surgical and functional outcome after resection of 64 petroclival meningiomas. JO - Cancers VL - 14 IS - 18 PY - 2022 SN - 2072-6694 ER - TY - JOUR AB - Ewing sarcoma (EwS) is a highly malignant sarcoma of bone and soft tissue with early metastatic spread and an age peak in early puberty. The prognosis in advanced stages is still dismal, and the long-term effects of established therapies are severe. Efficacious targeted therapies are urgently needed. Our previous work has provided preliminary safety and efficacy data utilizing T cell receptor (TCR) transgenic T cells, generated by retroviral gene transfer, targeting HLA-restricted peptides on the tumor cell derived from metastatic drivers. Here, we compared T cells engineered with either CRISPR/Cas9 or retroviral gene transfer. Firstly, we confirmed the feasibility of the orthotopic replacement of the endogenous TCR by CRISPR/Cas9 with a TCR targeting our canonical metastatic driver chondromodulin-1 (CHM1). CRISPR/Cas9-engineered T cell products specifically recognized and killed HLA-A*02:01+ EwS cell lines. The efficiency of retroviral transduction was higher compared to CRISPR/Cas9 gene editing. Both engineered T cell products specifically recognized tumor cells and elicited cytotoxicity, with CRISPR/Cas9 engineered T cells providing prolonged cytotoxic activity. In conclusion, T cells engineered with CRISPR/Cas9 could be feasible for immunotherapy of EwS and may have the advantage of more prolonged cytotoxic activity, as compared to T cells engineered with retroviral gene transfer. AU - Xue, B.* AU - von Heyking, K.* AU - Gassmann, H.* AU - Poorebrahim, M.* AU - Thiede, M.* AU - Schober, K.* AU - Mautner, J. AU - Hauer, J.* AU - Ruland, J.* AU - Busch, D.H.* AU - Thiel, U.* AU - Burdach, S.E.G.* C1 - 66832 C2 - 53306 TI - T cells directed against the metastatic driver chondromodulin-1 in ewing sarcoma: Comparative engineering with CRISPR/Cas9 vs. retroviral gene transfer for adoptive transfer. JO - Cancers VL - 14 IS - 22 PY - 2022 SN - 2072-6694 ER - TY - JOUR AB - Barrett's esophagus (BE) is the precursor of esophageal adenocarcinoma (EAC). Dysplastic BE (DBE) has a higher progression risk to EAC compared to non-dysplastic BE (NDBE). However, the miss rates for the endoscopic detection of DBE remain high. Fluorescence molecular endoscopy (FME) can detect DBE and mucosal EAC by highlighting the tumor-specific expression of proteins. This study aimed to identify target proteins suitable for FME. Publicly available RNA expression profiles of EAC and NDBE were corrected by functional genomic mRNA (FGmRNA) profiling. Following a class comparison between FGmRNA profiles of EAC and NDBE, predicted, significantly upregulated genes in EAC were prioritized by a literature search. Protein expression of prioritized genes was validated by immunohistochemistry (IHC) on DBE and NDBE tissues. Near-infrared fluorescent tracers targeting the proteins were developed and evaluated ex vivo on fresh human specimens. In total, 1976 overexpressed genes were identified in EAC (n = 64) compared to NDBE (n = 66) at RNA level. Prioritization and IHC validation revealed SPARC, SULF1, PKCι, and DDR1 (all p < 0.0001) as the most attractive imaging protein targets for DBE detection. Newly developed tracers SULF1-800CW and SPARC-800CW both showed higher fluorescence intensity in DBE tissue compared to paired non-dysplastic tissue. This study identified SPARC, SULF1, PKCι, and DDR1 as promising targets for FME to differentiate DBE from NDBE tissue, for which SULF1-800CW and SPARC-800CW were successfully ex vivo evaluated. Clinical studies should further validate these findings. AU - Zhao, X.* AU - Gabriels, R.Y.* AU - Hooghiemstra, W.T.R.* AU - Koller, M.* AU - Meersma, G.J.* AU - Buist-Homan, M.* AU - Visser, L.* AU - Robinson, D.J.* AU - Tenditnaya, A. AU - Gorpas, D. AU - Ntziachristos, V. AU - Karrenbeld, A.* AU - Kats-Ugurlu, G.* AU - Fehrmann, R.S.N.* AU - Nagengast, W.B.* C1 - 65409 C2 - 52284 TI - Validation of novel molecular imaging targets identified by functional genomic mRNA profiling to detect dysplasia in Barrett's Esophagus. JO - Cancers VL - 14 IS - 10 PY - 2022 SN - 2072-6694 ER - TY - JOUR AB - Background: Resection of jugular foramen schwannomas (JFSs) with minimal cranial nerve (CN) injury remains difficult. Reoperations in this vital region are associated with severe CN deficits. Methods: We performed a retrospective analysis at a tertiary neurosurgical center of patients who underwent surgery for JFSs between June 2007 and May 2020. We included nine patients (median age 60 years, 77.8% female, 22.2% male). Preoperative symptoms included hearing loss (66.6%), headache (44.4%), hoarseness (33.3%), dysphagia (44.4%), hypoglossal nerve palsy (22.2%), facial nerve palsy (33.3%), extinguished gag reflex (22.2%), and cerebellar dysfunction (44.4%). We observed Type A, B, C, and D tumors in 3, 1, 1, and 4 patients, respectively. A total of 77.8% (7/9) underwent a retrosigmoid approach, and 33.3% (3/9) underwent an extreme lateral infrajugular transcondylar (ELITE) approach. Gross total resection (GTR) was achieved in all cases. The rate of shunt-dependent hydrocephalus was 22.2% (2/9). No further complications requiring surgical intervention occurred during follow-up. The median follow-up time was 16.5 months (range 3–84 months). Conclusions: Considering the satisfying outcome, the GTR of JFSs is feasible in performing well-known skull base approaches. Additional invasive and complicated approaches were not needed. Radiosurgery may be an effective alternative for selected patients. AU - Aftahy, A.K.* AU - Groll, M.* AU - Barz, M.* AU - Bernhardt, D.* AU - Combs, S.E. AU - Meyer, B.* AU - Negwer, C.* AU - Gempt, J.* C1 - 62845 C2 - 51102 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Surgical management of jugular foramen schwannomas. JO - Cancers VL - 13 IS - 16 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - Head and neck squamous cell carcinomas (HNSCC) are common tumors with a poor overall prognosis. Poor survival is resulting from limited response to multi-modal therapy, high incidence of metastasis, and local recurrence. Treatment includes surgery, radio(chemo)therapy, and targeted therapy specific for EGFR and immune checkpoint inhibition. The understanding of the molecular basis for the poor outcome of HNSCC was improved using multi-OMICs approaches, which revealed a strong degree of inter-and intratumor heterogeneity (ITH) at the level of DNA mu-tations, transcriptome, and (phospho)proteome. Single-cell RNA-sequencing (scRNA-seq) identified RNA-expression signatures related to cell cycle, cell stress, hypoxia, epithelial differentiation, and a partial epithelial-to-mesenchymal transition (pEMT). The latter signature was correlated to nodal involvement and adverse clinical features. Mechanistically, shifts towards a mesenchymal phenotype equips tumor cells with migratory and invasive capacities and with an enhanced resistance to standard therapy. Hence, gradual variations of EMT as observed in HNSCC represent a potent driver of tumor progression that could open new paths to improve the stratification of patients and to innovate approaches to break therapy resistance. These aspects of molecular heterogeneity will be discussed in the present review. AU - Baumeister, P. AU - Zhou, J.* AU - Canis, M.* AU - Gires, O. C1 - 63377 C2 - 51305 TI - Epithelial-to-mesenchymal transition-derived heterogeneity in head and neck squamous cell carcinomas. JO - Cancers VL - 13 IS - 21 PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - The current study aims to determine whether exclusion of lung tissue from planning treatment volume (PTV) is a valid organ at risk (OAR)-sparing technique during internal mammary irradiation (IMNI). Twenty patients with left-sided breast cancer undergoing adjuvant radiotherapy including IMNI after mastectomy or lumpectomy with daily ConeBeam CT (CBCT; median n = 28) were enrolled in the current study. The daily dose distribution of the patients was estimated by recalculating treatment plans on CBCT-scans based on a standard PTV (PTV margin: 5mm-STD) and a modified PTV, which excluded overlapping lung tissue (ExLung). Using 3D-deformable dose accumulation, the dose coverage in the target volume was estimated in dependence of the PTV-margins. The estimated delivered dose in the IMN-CTV was significantly lower for the ExLung PTV compared to the STD PTV: ExLung: V95%: 76.6 ± 22.9%; V90%: 89.6 ± 13.2%, STD: V95%: 95.6 ± 7.4%; V90%: 99.1 ± 2.7%. Daily CBCT imaging cannot sufficiently compensate the anatomic changes and intrafraction movement throughout the treatment. Therefore, to ensure adequate delivery of the prescribed dose to the IMN-CTV, exclusion of lung tissue from the PTV to spare the OARs is not recommended. AU - Borm, K.J.* AU - Hofmann, C.* AU - Düsberg, M.* AU - Oechsner, M.* AU - Dapper, H.* AU - Devecka, M.* AU - Combs, S.E. C1 - 61871 C2 - 50491 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Excluding lung tissue from the ptv during internal mammary irradiation. A safe technique for oar-sparing? JO - Cancers VL - 13 IS - 8 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - The current study aims to determine whether hypofractionated radiotherapy (HF) leads to lower rates of acute radiodermatitis compared to conventional normofractionated radiotherapy (CF). A total of 166 patients with invasive breast cancer or DCIS were included in a prospective cohort study. Evaluation of acute radiodermatitis was obtained before radiotherapy, at the end of the treatment (T1), and 6 weeks after the treatment (T2) using CTCAE (v5.0) scores, the Skindex-16 questionnaire, and ultrasound measurement of the skin. CTCAE and Skindex-16 scores in the CF-group were significantly higher compared to the HF group indicating more pronounced side effects at the end of the treatment (CTCAE: CF-RT 1.0 (IQR: 0.0) vs. HF-RT 0.0 (0.25); p = 0.03; Skindex-16: CF: 20.8 (IQR: 25.8); HF: 8.3 (27.1); p = 0.04). At 6 weeks after the treatment, no significant differences between the two fractionation schemes were observed. Ultrasound based assessment showed that the skin thickness in the treated breast was higher compared to the healthy breast at all time-points. However, no significant difference between HF and CF was seen either at T1 or T2. The current study complements and confirms pre-existing evidence that HF leads to a lower degree of acute radiodermatitis and better patient reported outcome compared to CF at the end of treatment. This should be considered whenever fractionation of adjuvant breast cancer treatment is being dis-cussed. AU - Borm, K.J.* AU - Vennekate, J.K.* AU - Vagedes, J.* AU - Islam, M.O.A.* AU - Duma, M.N.* AU - Loos, M.* AU - Combs, S.E. AU - Schiller, K.* AU - Klusen, S.* AU - Paepke, S.* AU - Kiechle, M.B.* AU - Paepke, D.* C1 - 63562 C2 - 51587 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - A comprehensive prospective comparison of acute skin toxicity after hypofractionated and normofractionated radiation therapy in breast cancer. JO - Cancers VL - 13 IS - 22 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - Transcription factors can serve as links between tumor microenvironment signaling and oncogenesis. Interferon regulatory factor 9 (IRF9) is recruited and expressed upon interferon stimulation and is dependent on cofactors that exert in tumor-suppressing or oncogenic functions via the JAK-STAT pathway. IRF9 is frequently overexpressed in human lung cancer and is associated with decreased patient survival; however, the underlying mechanisms remain to be elucidated. Here, we used stably transduced lung adenocarcinoma cell lines (A549 and A427) to overexpress or knockdown IRF9. Overexpression led to increased oncogenic behavior in vitro, including enhanced proliferation and migration, whereas knockdown reduced these effects. These findings were confirmed in vivo using lung tumor xenografts in nude mice, and effects on both tumor growth and tumor mass were observed. Using RNA sequencing, we identified versican (VCAN) as a novel downstream target of IRF9. Indeed, IRF9 and VCAN expression levels were found to be correlated. We showed for the first time that IRF9 binds at a newly identified response element in the promoter region of VCAN to regulate its transcription. Using an siRNA approach, VCAN was found to enable the oncogenic properties (proliferation and migration) of IRF9 transduced cells, perhaps with CDKN1A involvement. The targeted inhibition of IRF9 in lung cancer could therefore be used as a new treatment option without multimodal interference in microenvironment JAK-STAT signaling. AU - Brunn, D.* AU - Turkowski, K.* AU - Günther, S.* AU - Weigert, A.* AU - Muley, T.* AU - Kriegsmann, M.* AU - Winter, H.* AU - Dammann, R.H.* AU - Stathopoulos, G.T. AU - Thomas, M.* AU - Guenther, A.* AU - Grimminger, F.* AU - Pullamsetti, S.S.* AU - Seeger, W.* AU - Savai, R.* C1 - 61021 C2 - 50017 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Interferon regulatory factor 9 promotes lung cancer progression via regulation of versican. JO - Cancers VL - 13 IS - 2 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal human cancers. Innovative treatment concepts may enhance oncological outcome. Clinically relevant tumor models are essential in developing new therapeutic strategies. In the present study, we used two human PDAC cell lines for an orthotopic xenograft mouse model and compared treatment characteristics between this in vivo tumor model and PDAC patients. Tumor-bearing mice received stereotactic high-precision irradiation using arc technique after 3D-treatment planning. Induction of DNA damage in tumors and organs at risk (OARs) was histopathologically analyzed by the DNA damage marker γH2AX and compared with results after unprecise whole-abdomen irradiation. Our mouse model and preclinical setup reflect the characteristics of PDAC patients and clinical RT. It was feasible to perform stereotactic high-precision RT after defining tumor and OARs by CT imaging. After stereotactic RT, a high rate of DNA damage was mainly observed in the tumor but not in OARs. The calculated dose distributions and the extent of the irradiation field correlate with histopathological staining and the clinical example. We established and validated 3D-planned stereotactic RT in an orthotopic PDAC mouse model, which reflects the human RT. The efficacy of the whole workflow of imaging, treatment planning, and high-precision RT was proven by longitudinal analysis showing a significant improved survival. Importantly, this model can be used to analyze tumor regression and therapy-related toxicity in one model and will allow drawing clinically relevant conclusions. AU - Dobiasch, S. AU - Kampfer, S.* AU - Steiger, K.* AU - Schilling, D. AU - Fischer, J.C.* AU - Schmid, T.E. AU - Weichert, W.* AU - Wilkens, J.J. AU - Combs, S.E. C1 - 63519 C2 - 51576 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Histopathological tumor and normal tissue responses after 3d-planned arc radiotherapy in an orthotopic xenograft mouse model of human pancreatic cancer. JO - Cancers VL - 13 IS - 22 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - Background: Mounting evidence shows that adiposity increases female-specific cancer risk, but the role of body fat distribution is less clear. We used a two-sample Mendelian randomization (MR) approach to elucidate causal relations of body fat distribution to the risks of breast, endometrial and ovarian cancers and their subtypes. Methods: Body composition was assessed using segmental bioelectrical impedance analysis, yielding trunk, arm, and leg fat ratios (TFR, AFR, LFR) and BMI including 195,043 and 434,794 European women, respectively. The sample sizes for the outcomes ranged between 58,396 and 228,951. Causal effects were estimated per one standard deviation increment in the respective exposure within the radial regression framework. Robust sensitivity analyses were performed to verify MR assumptions. In a multivariable MR setting, the proportion of risk attributable to overall and abdominal fat content was assessed. Results: TFR, which represents abdominal fat content, was associated with ovarian cancer and its clear cell and endometrioid histotypes independent of overall fat content. BMI was inversely associated with breast cancer and its ER− and ER+ subtypes, but positively with endometrial cancer and ovarian cancer, including its endometrioid histotype. These estimates were confirmed using AFR as proxy for overall body fat. Conclusions: Visceral adiposity seems to be a driver of elevated ovarian cancer risk, particularly of the endometrioid and clear cell ovarian cancer histotypes. General adiposity de-creases the risk of breast cancer but increases the risk of endometrial and ovarian cancer. AU - Freuer, D.* AU - Linseisen, J. AU - O’mara, T.A.* AU - Leitzmann, M.* AU - Baurecht, H.* AU - Baumeister, S.E.* AU - Meisinger, C.* C1 - 63258 C2 - 51412 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Body fat distribution and risk of breast, endometrial, and ovarian cancer: A two-sample mendelian randomization study. JO - Cancers VL - 13 IS - 20 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - Interleukin-1 cytokines are key proinflammatory cytokines which have been implicated with differing pro- and antitumorigenic properties. Recent years have brought exciting insights and developments in IL-1-targeted therapies. Here, we present an overview of past and present research focusing on the role of IL-1 in cancer, with a special focus on clinical research and on therapeutic implications. With this, we strive to assist scientists in their future research objectives and to highlight possible directions for IL-1-targeting therapies in the coming years. Since its discovery, interleukin-1 has been extensively studied in a wide range of medical fields. Besides carrying out vital physiological functions, it has been implicated with a pivotal role in the progression and spreading of different cancer entities. During the last years, several clinical trials have been conducted, shedding light on the role of IL-1 blocking agents for the treatment of cancer. Additionally, recent developments in the field of immuno-oncology have implicated IL-1- induced signaling cascades as a major driver of severe chimeric antigen receptor T cell-associated toxicities such as cytokine release syndrome and immune effector cell-associated neurotoxicity. In this review, we summarize current clinical trials investigating the role of IL-1 blockade in cancer treatment and elaborate the proposed mechanism of these innovative treatment approaches. Additionally, we highlight cutting-edge developments utilizing IL-1 blocking agents to enhance the safety and efficacy of adoptive T cell therapy. AU - Gottschlich, A.* AU - Endres, S. AU - Kobold, S. C1 - 61142 C2 - 50302 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Therapeutic strategies for targeting IL-1 in cancer. JO - Cancers VL - 13 IS - 3 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - Invasive nonfunctioning pituitary tumors (NFPTs) are non-resectable neoplasms associated with frequent relapse and significant comorbidities. Current treatments, including somatostatin receptor 2 (SSTR2)-directed somatostatin analogs (SSAs), often fail against NFPTs. Thus, identifying effective therapies is clinically relevant. As NFPTs express SSTR3 at high levels, pasireotide, a multireceptor-targeted SSA, might be beneficial. Here we evaluated pasireotide in the only representative model of spontaneous NFPTs (MENX rats) in vivo. Octreotide long-acting release (LAR), pasireotide LAR, or placebo, were administered to age-matched, tumor-bearing MENX rats of both sexes for 28 d or 56 d. Longitudinal high-resolution magnetic resonance imaging monitored tumor growth. While tumors in placebo-treated rats increased in volume over time, PTs in drug-treated rats displayed significant growth suppression, and occasional tumor shrinkage. Pasireotide elicited stronger growth inhibition. Radiological responses correlated with tumors’ proliferation rates. Both SSAs, but especially pasireotide, were more effective in female vs. male rats. Basal Sstr3 expression was significantly higher in the former group. It is noteworthy that female human NFPTs patients also have a trend towards higher SSTR3 expression. Altogether, our studies provide the rationale for testing pasireotide in patients with residual/recurrent NFPTs. If confirmed, the sex-related SSTR3 expression might be used as criteria to stratify NFPTs patients for treatment with pasireotide. AU - Gulde, S. AU - Wiedemann, T. AU - Schillmaier, M.* AU - Valença, I. AU - Lupp, A.* AU - Steiger, K.* AU - Yen, H.Y.* AU - Bäuerle, S.* AU - Notni, J.* AU - Luque, R.* AU - Schmid, H.* AU - Schulz, S.* AU - Ankerst, D.P.* AU - Schilling, F.* AU - Pellegata, N.S. C1 - 62359 C2 - 50804 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Gender-specific efficacy revealed by head-to-head comparison of pasireotide and octreotide in a representative in vivo model of nonfunctioning pituitary tumors. JO - Cancers VL - 13 IS - 12 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - Brain metastases are the most common intracranial malignant tumor in adults and are a cause of significant morbidity and mortality for cancer patients. Large brain metastases, defined as tumors with a maximum dimension >2 cm, present a unique clinical challenge for the delivery of stereotactic radiosurgery (SRS) as patients often present with neurologic symptoms that require expeditious treatment that must also be balanced against the potential consequences of surgery and radiation therapy—namely, leptomeningeal disease (LMD) and radionecrosis (RN). Hypofractionated stereotactic radiotherapy (HSRT) and pre-operative SRS have emerged as novel treatment techniques to help improve local control rates and reduce rates of RN and LMD for this patient population commonly managed with post-operative SRS. Recent literature suggests that pre-operative SRS can potentially half the risk of LMD compared to post-operative SRS and that HSRT can improve risk of RN to less than 10% while improving local control when meeting the appropriate goals for biologically effective dose (BED) and dose-volume constraints. We recommend a 3-or 5-fraction regimen in lieu of SRS delivering 15 Gy or less for large metastases or resection cavities. We provide a table comparing the BED of commonly used SRS and HSRT regimens, and provide an algorithm to help guide the management of these challenging clinical scenarios. AU - Gutschenritter, T.* AU - Venur, V.A.* AU - Combs, S.E. AU - Vellayappan, B.* AU - Patel, A.P.* AU - Foote, M.* AU - Redmond, K.J.* AU - Wang, T.J.C.* AU - Sahgal, A.* AU - Chao, S.T.* AU - Suh, J.H.* AU - Chang, E.L.* AU - Ellenbogen, R.G.* AU - Lo, S.S.* C1 - 60900 C2 - 49660 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - The judicious use of stereotactic radiosurgery and hypofractionated stereotactic radiotherapy in the management of large brain metastases. JO - Cancers VL - 13 IS - 1 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - Many countries have reported survival inequalities due to regional socioeconomic deprivation. To quantify the potential gain from eliminating cancer survival disadvantages associated with area-based deprivation in Germany, we calculated the number of avoidable excess deaths. We used population-based cancer registry data from 11 of 16 German federal states. Patients aged ≥15 years diagnosed with an invasive malignant tumor between 2008 and 2017 were included. Area-based socioeconomic deprivation was assessed using the quintiles of the German Index of Multiple Deprivation (GIMD) 2010 on a municipality level nationwide. Five-year age-standardized relative survival for 25 most common cancer sites and for total cancer were calculated using period analysis. Incidence and number of avoidable excess deaths in Germany in 2013-2016 were estimated. Summed over the 25 cancer sites, 4100 annual excess deaths (3.0% of all excess deaths) could have been avoided each year in Germany during the period 2013-2016 if relative survival were in all regions comparable with the least deprived regions. Colorectal, oral and pharynx, prostate, and bladder cancer contributed the largest numbers of avoidable excess deaths. Our results provide a good basis to estimate the potential of intervention programs for reducing socioeconomic inequalities in cancer burden in Germany. AU - Jansen, L.* AU - Kanbach, J.* AU - Finke, I.* AU - Arndt, V.* AU - Emrich, K.* AU - Holleczek, B.* AU - Kajüter, H.* AU - Kieschke, J.* AU - Maier, W. AU - Pritzkuleit, R.* AU - Sirri, E.* AU - Schwettmann, L. AU - Erb, C.* AU - Brenner, H.* C1 - 61070 C2 - 50029 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Estimation of the potentially avoidable excess deaths associated with socioeconomic inequalities in cancer survival in Germany. JO - Cancers VL - 13 IS - 2 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - Purpose: We report the outcome of a mono-institutional retrospective study of sinonasal carcinoma with the primary focus on GTV (gross tumor volume) and the effect of radiotherapy. Methods: 53 patients with sinonasal carcinoma and that of the nasal cavity, paranasal sinus or both except lymphoma were included. All patients were treated between 1999 and 2017. For tumor volume delineation, all pre-therapeutic images were fused to the planning CT (computed tomography). Results: The median follow-up was 17 months [0.3–60], the median age 60 years, 35 males and 18 females were included. Squamous cell carcinoma (SCC) (60.4%) was the predominant histology, followed by adenocarcinoma (15.1%). The mean composite OS (overall survival) time was 33.3 ± 3.5 months. There was no significant difference in the 5 y composite OS between tumor localization or radiotherapy setting. The simultaneous integrated boost concept showed a trend towards improving five-year composite OS compared to the sequential boost concept. The only factor with a significant impact on the 5 y composite OS rate was the pre-therapeutic GTV (cutoff 75 cm ; p = 0.033). The GTV ≥ 100 cm has no effect on the 5 y composite OS rate for SCC. Conclusions: The pre-therapeutic GTV is a prognostic factor for five-year composite OS for the entire group of patients with sinonasal tumors, influencing the outcome after completion of all treatment strategies. The GTV seems to not influence five-year composite OS in SCC. For this rare tumor entity, an intensive, multidisciplinary discussion is essential to finding the best treatment option for the patient. 3 3 AU - Klymenko, O.* AU - Buchberger, A.M.S.* AU - Wollenberg, B.* AU - Wolff, K.D.* AU - Kehl, V.* AU - Combs, S.E. AU - Pickhard, A.* AU - Pigorsch, S.U.* C1 - 62051 C2 - 50610 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Radiooncological view on therapy outcome after multidisciplinary treatment of sinonasal tumors. JO - Cancers VL - 13 IS - 10 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - We investigated the potential of respiratory gating to mitigate the motion-caused misdos-age in lung stereotactic body radiotherapy (SBRT). For fourteen patients with lung tumors, we investigated treatment plans for a gating window (GW) including three breathing phases around the maximum exhalation phase, GW40-60. For a subset of six patients, we also assessed a preceding three-phase GW20-40 and six-phase GW20-70. We analyzed the target volume, lung, esophagus, and heart doses. Using normal tissue complication probability (NTCP) models, we estimated radiation pneumonitis and esophagitis risks. Compared to plans without gating, GW40-60 significantly reduced doses to organs at risk without impairing the tumor doses. On average, the mean lung dose decreased by 0.6 Gy (p < 0.001), treated lung V20Gy by 2.4% (p = 0.003), esophageal dose to 5cc by 2.0 Gy (p = 0.003), and maximum heart dose by 3.2 Gy (p = 0.009). The model-estimated mean risks of 11% for pneumonitis and 12% for esophagitis without gating decreased upon GW40-60 to 7% and 9%, respectively. For the highest-risk patient, gating reduced the pneumonitis risk from 43% to 32%. Gating is most beneficial for patients with high-toxicity risks. Pre-treatment toxicity risk assessment may help optimize patient selection for gating, as well as GW selection for individual patients. AU - Kraus, K.M. AU - Simonetto, C. AU - Kundrát, P.* AU - Waitz, V.* AU - Borm, K.J.* AU - Combs, S.E. C1 - 63309 C2 - 51468 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Potential morbidity reduction for lung stereotactic body radiation therapy using respiratory gating. JO - Cancers VL - 13 IS - 20 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - Infection with the human papillomavirus (HPV) has been identified as a major risk factor for oropharyngeal cancer (OPC). HPV-related OPCs have been shown to be more radiosensitive and to have a reduced risk for cancer related death. Hence, the histological determination of HPV status of cancer patients depicts an essential diagnostic factor. We investigated the ability of deep learning models for imaging based HPV status detection. To overcome the problem of small medical datasets, we used a transfer learning approach. A 3D convolutional network pre-trained on sports video clips was fine-tuned, such that full 3D information in the CT images could be exploited. The video pre-trained model was able to differentiate HPV-positive from HPV-negative cases, with an area under the receiver operating characteristic curve (AUC) of 0.81 for an external test set. In comparison to a 3D convolutional neural network (CNN) trained from scratch and a 2D architecture pre-trained on ImageNet, the video pre-trained model performed best. Deep learning models are capable of CT image-based HPV status determination. Video based pre-training has the ability to improve training for 3D medical data, but further studies are needed for verification. AU - Lang, D.M. AU - Peeken, J.C. AU - Combs, S.E. AU - Wilkens, J.J.* AU - Bartzsch, S. C1 - 61377 C2 - 50181 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Deep learning based HPV status prediction for oropharyngeal cancer patients. JO - Cancers VL - 13 IS - 4 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - High‐Z gold nanoparticles (AuNPs) conjugated to a targeting antibody can help to improve tumor control in radiotherapy while simultaneously minimizing radiotoxicity to adjacent healthy tissue. This paper summarizes the main findings of a joint research program which applied AuNP‐conjugates in preclinical modeling of radiotherapy at the Klinikum rechts der Isar, Technical University of Munich and Helmholtz Zentrum München. A pharmacokinetic model of superparamagnetic iron oxide nanoparticles was developed in preparation for a model simulating. AU - Li, W.B. AU - Stangl, S.* AU - Klapproth, A. AU - Shevtsov, M.* AU - Hernandez, A.* AU - Kimm, M.A.* AU - Schuemann, J.* AU - Qiu, R.* AU - Michalke, B. AU - Bernal, M.A.* AU - Li, J.* AU - Hürkamp, K. AU - Zhang, Y.* AU - Multhoff, G.* C1 - 63376 C2 - 51511 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Application of high‐Z gold nanoparticles in targeted cancer radiotherapy—pharmacokinetic modeling, Monte Carlo simulation and radiobiological effect modeling. JO - Cancers VL - 13 IS - 21 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - Small cell lung carcinoma (SCLC) is a highly aggressive malignancy with a very high mortality rate. A prominent part of this is because these carcinomas are refractory to chemotherapies, such as etoposide or cisplatin, making effective treatment almost impossible. Here, we report that elevated expression of the RAGE variant-V in SCLC promotes homology-directed DNA DSBs repair when challenged with anti-cancer drugs. This variant exclusively localizes to the nucleus, interacts with members of the double-strand break (DSB) repair machinery and thus promotes the recruitment of DSBs repair factors at the site of damage. Increased expression of this variant thus, promotes timely DNA repair. Congruently, the tumor cells expressing high levels of variant-V can tolerate chemotherapeutic drug treatment better than the RAGE depleted cells. Our findings reveal a yet undisclosed role of the RAGE variant-V in the homology-directed DNA repair. This variant thus can be a potential target to be considered for future therapeutic approaches in advanced SSLC. AU - Madhavan, B.K.* AU - Han, Z.* AU - Singh, B.* AU - Bordt, N.* AU - Kaymak, S.* AU - Bandapalli, O.R.* AU - Kihm, L.* AU - Shahzad, K.* AU - Isermann, B.* AU - Herzig, S. AU - Nawroth, P.P.* AU - Kumar, V.* C1 - 62229 C2 - 50746 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Elevated expression of the rage variant-v in sclc mitigates the effect of chemotherapeutic drugs. JO - Cancers VL - 13 IS - 11 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - In contrast with other strategies, immunotherapy is the only treatment aimed at empowering the immune system to increase the response against tumor growth. Immunotherapy has a role in the treatment of bladder cancer (BC) due to these tumors' high tumor mutational burden (TMB) and mostly prominent immune infiltrate. The therapy or combination has to be adjusted to the tumor's immunobiology. Recently, a new class of immunotherapeutic agents, immune checkpoint inhibitors (ICI), has shown potential in increasing treatment chances for patients with genitourinary cancers, improving their oncological outcomes. The clinical efficacy of ICI has been shown in both the first-line treatment of cisplatin-ineligible patients, with programmed death ligand 1 (PD-L1)-positive tumors (atezolizumab, pembrolizumab), and in second-line settings, for progression after platinum-based chemotherapy (atezolizumab, pembrolizumab, and nivolumab for FDA and EMA; durvalumab and avelumab for FDA alone). Predicting the response to ICI is important since only a subset of patients undergoing ICI therapy develop a concrete and lasting response. Most of the patients require a different therapy or therapy combination to achieve tumor control. The cancer immunity cycle provides a conceptual framework to assist therapy selection. Biomarkers to predict response to ICI must identify where the cancer immunity cycle is disrupted. We reviewed the current knowledge on ICI treatment in BC, going from basic science to current data and available clinical evidence. Secondly, a critical analysis of published data is provided, and an original panel of biomarkers able to predict response to ICI treatment, based on tumor-specific immune profiling, is proposed. AU - Mancini, M.* AU - Righetto, M.* AU - Nößner, E. C1 - 63795 C2 - 50414 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Checkpoint inhibition in bladder cancer: Clinical expectations, current evidence, and proposal of future strategies based on a tumor-specific immunobiological approach. JO - Cancers VL - 13 IS - 23 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - Patient-derived induced pluripotent stem cells (iPSCs) provide a unique platform to study hereditary disorders and predisposition syndromes by resembling germline mutations of affected individuals and by their potential to differentiate into nearly every cell type of the human body. We employed plucked human hair from two siblings with a family history of cancer carrying a pathogenic CDKN2A variant, P16-p.G101W/P14-p.R115L, to generate patient-specific iPSCs in a cancer-prone ancestry for downstream analytics. The differentiation capacity to pancreatic progenitors and to pancreatic duct-like organoids (PDLOs) according to a recently developed protocol remained unaffected. Upon inducible expression of KRASG12D using a piggyBac transposon system in CDKN2A-mutated PDLOs, we revealed structural and molecular changes in vitro, including disturbed polarity and epithelial-to-mesenchymal (EMT) transition. CDKN2A-mutated KRASG12D PDLO xenotransplants formed either a high-grade precancer lesion or a partially dedifferentiated PDAC-like tumor. Intriguingly, P14/P53/P21 and P16/RB cell-cycle checkpoint controls have been only partly overcome in these grafts, thereby still restricting the tumorous growth. Hereby, we provide a model for hereditary human pancreatic cancer that enables dissection of tumor initiation and early development starting from patient-specific CDKN2A-mutated pluripotent stem cells. AU - Merkle, J.* AU - Breunig, M.* AU - Schmid, M.* AU - Allgöwer, C.* AU - Krüger, J.* AU - Melzer, M.K.* AU - Bens, S.* AU - Siebert, R.* AU - Perkhofer, L.* AU - Azoitei, N.* AU - Seufferlein, T.* AU - Heller, S.* AU - Meier, M. AU - Müller, M.* AU - Kleger, A.* AU - Hohwieler, M.* C1 - 63326 C2 - 51471 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Cdkn2a-mutated pancreatic ductal organoids from induced pluripotent stem cells to model a cancer predisposition syndrome. JO - Cancers VL - 13 IS - 20 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - BACKGROUND: Pseudohypoxic tumors activate pro-oncogenic pathways typically associated with severe hypoxia even when sufficient oxygen is present, leading to highly aggressive tumors. Prime examples are pseudohypoxic pheochromocytomas and paragangliomas (p-PPGLs), neuroendendocrine tumors currently lacking effective therapy. Previous attempts to generate mouse models for p-PPGLs all failed. Here, we describe that the rat MENX line, carrying a Cdkn1b (p27) frameshift-mutation, spontaneously develops pseudohypoxic pheochromocytoma (p-PCC). METHODS: We compared rat p-PCCs with their cognate human tumors at different levels: histology, immunohistochemistry, catecholamine profiling, electron microscopy, transcriptome and metabolome. The vessel architecture and angiogenic potential of pheochromocytomas (PCCs) was analyzed by light-sheet fluorescence microscopy ex vivo and multi-spectral optoacoustic tomography (MSOT) in vivo. RESULTS: The analysis of tissues at various stages, from hyperplasia to advanced grades, allowed us to correlate tumor characteristics with progression. Pathological changes affecting the mitochrondrial ultrastructure where present already in hyperplasias. Rat PCCs secreted high levels of norepinephrine and dopamine. Transcriptomic and metabolomic analysis revealed changes in oxidative phosphorylation that aggravated over time, leading to an accumulation of the oncometabolite 2-hydroxyglutarate, and to hypermethylation, evident by the loss of the epigenetic mark 5-hmC. While rat PCC xenografts showed high oxygenation, induced by massive neoangiogenesis, rat primary PCC transcriptomes possessed a pseudohypoxic signature of high Hif2a, Vegfa, and low Pnmt expression, thereby clustering with human p-PPGL. CONCLUSION: Endogenous rat PCCs recapitulate key phenotypic features of human p-PPGLs. Thus, MENX rats emerge as the best available animal model of these aggressive tumors. Our study provides evidence of a link between cell cycle dysregulation and pseudohypoxia. AU - Mohr, H. AU - Ballke, S.* AU - Bechmann, N.* AU - Gulde, S. AU - Malekzadeh Najafabadi, J. AU - Peitzsch, M.* AU - Ntziachristos, V. AU - Steiger, K.* AU - Wiedemann, T. AU - Pellegata, N.S. C1 - 60950 C2 - 50001 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Mutation of the cell cycle regulator p27kip1 drives pseudohypoxic pheochromocytoma development. JO - Cancers VL - 13 IS - 1 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - Purpose: To evaluate the suitability of psoas and erector spinae muscle proton density fat fraction (PDFF) and fat volume as biomarkers for monitoring cachexia severity in an oncological cohort, and to evaluate regional variances in muscle parameters over time. Methods: In this prospective study, 58 oncological patients were examined by a 3 T MRI receiving between one and five scans. Muscle volume and PDFF were measured, segmentation masks were divided into proximal, middle and distal muscle section. Results: A regional variation of fat distribution in erector spinae muscle at baseline was found (p < 0.01). During follow‐ups significant relative change of muscle parameters was observed. Relative maximum change of erector spinae muscle showed a significant regional variation. Correlation testing with age as a covariate revealed significant correlations for baseline psoas fat volume (r = −0.55, p < 0.01) and baseline psoas PDFF (r = −0.52, p = 0.02) with maximum BMI change during the course of the disease. Conclusion: In erector spinae muscles, a regional variation of fat distribution at baseline and relative maximum change of muscle parameters was observed. Our results indicate that psoas muscle PDFF and fat volume could serve as MRI-determined biomarkers for early risk stratification and disease monitoring regarding progression and severity of weight loss in cancer cachexia. AU - Patzelt, L.* AU - Junker, D.* AU - Syväri, J.* AU - Burian, E.* AU - Wu, M.* AU - Prokopchuk, O.* AU - Nitsche, U.* AU - Makowski, M.R.* AU - Braren, R.F.* AU - Herzig, S. AU - Diaz, M.B. AU - Karampinos, D.C.* C1 - 62931 C2 - 51174 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Mri‐determined psoas muscle fat infiltration correlates with severity of weight loss during cancer cachexia. JO - Cancers VL - 13 IS - 17 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - Background: In patients with soft-tissue sarcomas of the extremities, the treatment decision is currently regularly based on tumor grading and size. The imaging-based analysis may pose an alternative way to stratify patients’ risk. In this work, we compared the value of MRI-based radiomics with expert-derived semantic imaging features for the prediction of overall survival (OS). Methods: Fat-saturated T2-weighted sequences (T2FS) and contrast-enhanced T1-weighted fatsaturated (T1FSGd) sequences were collected from two independent retrospective cohorts (training: 108 patients; testing: 71 patients). After preprocessing, 105 radiomic features were extracted. Semantic imaging features were determined by three independent radiologists. Three machine learning techniques (elastic net regression (ENR), least absolute shrinkage and selection operator, and random survival forest) were compared to predict OS. Results: ENR models achieved the best predictive performance. Histologies and clinical staging differed significantly between both cohorts. The semantic prognostic model achieved a predictive performance with a C-index of 0.58 within the test set. This was worse compared to a clinical staging system (C-index: 0.61) and the radiomic models (C-indices: T1FSGd: 0.64, T2FS: 0.63). Both radiomic models achieved significant patient stratification. Conclusions: T2FS and T1FSGd-based radiomic models outperformed semantic imaging features for prognostic assessment. AU - Peeken, J.C. AU - Neumann, J.* AU - Asadpour, R.* AU - Leonhardt, Y.* AU - Moreira, J.R.* AU - Hippe, D.S.* AU - Klymenko, O.* AU - Foreman, S.C.* AU - von Schacky, C.E.* AU - Spraker, M.B.* AU - Schaub, S.K.* AU - Dapper, H.* AU - Knebel, C.* AU - Mayr, N.A.* AU - Woodruff, H.C.* AU - Lambin, P.* AU - Nyflot, M.J.* AU - Gersing, A.S.* AU - Combs, S.E. C1 - 61869 C2 - 50489 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Prognostic assessment in high-grade soft-tissue sarcoma patients: A comparison of semantic image analysis and radiomics. JO - Cancers VL - 13 IS - 8 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - The purpose of this study is to use a multi-technique approach to detect the effects of spatially fractionated X-ray Microbeam (MRT) and Minibeam Radiation Therapy (MB) and to compare them to seamless Broad Beam (BB) irradiation. Healthy-and Glioblastoma (GBM)-bearing male Fischer rats were irradiated in-vivo on the right brain hemisphere with MRT, MB and BB delivering three different doses for each irradiation geometry. Brains were analyzed post mortem by multi-scale X-ray Phase Contrast Imaging–Computed Tomography (XPCI-CT), histology, immunohistochemistry, X-ray Fluorescence (XRF), Small-and Wide-Angle X-ray Scattering (SAXS/WAXS). XPCI-CT discriminates with high sensitivity the effects of MRT, MB and BB irradiations on both healthy and GBM-bearing brains producing a first-time 3D visualization and morphological analysis of the radio-induced lesions, MRT and MB induced tissue ablations, the presence of hyperdense deposits within specific areas of the brain and tumor evolution or regression with respect to the evaluation made few days post-irradiation with an in-vivo magnetic resonance imaging session. Histology, immunohistochemistry, SAXS/WAXS and XRF allowed identification and classification of these deposits as hydroxyapatite crystals with the coexistence of Ca, P and Fe mineralization, and the multi-technique approach enabled the realization, for the first time, of the map of the differential radiosensitivity of the different brain areas treated with MRT and MB. 3D XPCI-CT datasets enabled also the quantification of tumor volumes and Ca/Fe deposits and their full-organ visualization. The multi-scale and multi-technique approach enabled a detailed visualization and classification in 3D of the radio-induced effects on brain tissues bringing new essential information towards the clinical implementation of the MRT and MB radiation therapy techniques. AU - Romano, M.* AU - Bravin, A.* AU - Mittone, A.* AU - Eckhardt, A.* AU - Barbone, G.E.* AU - Sancey, L.* AU - Dinkel, J.* AU - Bartzsch, S. AU - Ricke, J.* AU - Alunni-Fabbroni, M.* AU - Hirner-Eppeneder, H.* AU - Karpov, D.* AU - Giannini, C.* AU - Bunk, O.* AU - Bouchet, A.* AU - Ruf, V.* AU - Giese, A.* AU - Coan, P.* C1 - 63203 C2 - 51382 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - A multi-scale and multi-technique approach for the characterization of the effects of spatially fractionated X-ray radiation therapies in a preclinical model. JO - Cancers VL - 13 IS - 19 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - The heightened energetic demand increases lactate dehydrogenase (LDH) activity, the corresponding oncometabolite lactate, expression of heat shock proteins (HSPs) and thereby promotes therapy resistance in many malignant tumor cell types. Therefore, we assessed the coregulation of LDH and the heat shock response with respect to radiation resistance in different tumor cells (B16F10 murine melanoma and LS174T human colorectal adenocarcinoma). The inhibition of LDH activity by oxamate or GNE-140, glucose deprivation and LDHA/B double knockout (LDH−/−) in B16F10 and LS174T cells significantly diminish tumor growth; ROS production and the cytosolic expression of different HSPs, including Hsp90, Hsp70 and Hsp27 concomitant with a reduction of heat shock factor 1 (HSF1)/pHSF1. An altered lipid metabolism mediated by a LDHA/B double knockout results in a decreased presence of the Hsp70-anchoring glycosphingolipid Gb3 on the cell surface of tumor cells, which, in turn, reduces the membrane Hsp70 density and increases the extracellular Hsp70 levels. Vice versa, elevated extracellular lactate/pyruvate concentrations increase the membrane Hsp70 expression in wildtype tumor cells. Functionally, an inhibition of LDH causes a generalized reduction of cytosolic and membrane-bound HSPs in tumor cells and significantly increases the radiosensitivity, which is associated with a G2/M arrest. We demonstrate that targeting of the lactate/pyruvate metabolism breaks the radioresistance by impairing the stress response. AU - Schwab, M.* AU - Thunborg, K.* AU - Azimzadeh, O. AU - von Toerne, C. AU - Werner, C.* AU - Shevtsov, M.* AU - Di Genio, T.* AU - Zdralevic, M.* AU - Pouysségur, J.* AU - Renner, K.* AU - Kreutz, M.* AU - Multhoff, G.* C1 - 62703 C2 - 50973 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Targeting cancer metabolism breaks radioresistance by impairing the stress response. JO - Cancers VL - 13 IS - 15 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - Human adipose-derived stem/stromal cells (ASCs) are increasingly used as auto-transplants in regenerative medicine to restore tissue defects or induce wound healing, especially in cancer pa-tients. The impact of ASCs on squamous cell carcinoma of the upper aerodigestive tract (UAT) including head and neck and esophageal squamous cell carcinoma (HNSCC and ESCC) is not yet fully understood. ASCs were cultured from subcutaneous, abdominal lipoaspirates of five patients, who received auto-transplants to the head and neck. Supernatants were tested for paracrine effects in functional in vitro assays of proliferation of HNSCC tumor cell line FaDu and ESCC cell line Kyse30, and their cell migration/invasion capacities in Boyden chambers, in addition to endothelial tube formation assay using human umbilical vein endothelial cells (HUVECs). All ASC-derived super-natants enhanced proliferation of FaDu cells, invasive migration, and tube formation by HUVECs, compared to controls. Of five patients’ lipoaspirates, ASC-derived supernatants of four patients increased proliferation and invasive migration in Kyse30 cells. The data suggests that ASCs can promote tumor cell proliferation, invasiveness, and neo-angiogenesis in these tumor cell lines of the UAT and HUVEC in a paracrine manner. Although clinical studies on the subject of oncological safety are still needed, these findings emphasize the importance of complete tumor removal before ASCs are used in the head and neck. AU - Sharaf, K.* AU - Eggersmann, T.K.* AU - Haider, S.P.* AU - Schwenk-Zieger, S.* AU - Zhou, J.* AU - Gires, O. AU - Lechner, A.* AU - Canis, M.* AU - Haubner, F.* C1 - 62171 C2 - 50676 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Human adipose-derived stem/stromal cells promote proliferation and migration in head and neck cancer cells. JO - Cancers VL - 13 IS - 11 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - Lung cancer is the leading cancer killer worldwide, imposing grievous challenges for patients and clinicians. The incidence of lung adenocarcinoma (LUAD), the main histologic subtype of lung cancer, is still increasing in current-, ex-, and even non-smokers, whereas its five-year survival rate is approximately 15% as the vast majority of patients usually present with advanced disease at the time of diagnosis. The generation of novel drugs targeting key disease driver mutations has created optimism for the treatment of LUAD, but, as these mutations are not universal, this therapeutic line benefits only a subset of patients. More recently, the advent of targeted immunotherapies and their documented clinical efficacy in many different cancers, including LUAD, have started to change cancer management. Immunotherapies have been developed in order to overcome the cancer’s ability to develop mechanisms of immune resistance, i.e., to adapt to and evade the host inflammatory and immune responses. Identifying a cancer’s immune resistance mechanisms will likely advance the development of personalized immunotherapies. This review examines the key pathways of immune resistance at play in LUAD and explores therapeutic strategies which can unleash potent antitumor immune responses and significantly improve therapeutic efficacy, quality of life, and survival in LUAD. AU - Spella, M.* AU - Stathopoulos, G.T. C1 - 61114 C2 - 50051 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Immune resistance in lung adenocarcinoma. JO - Cancers VL - 13 IS - 3 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - Microbeam radiotherapy (MRT) is a preclinical method of delivering spatially-fractionated radiotherapy aiming to improve the therapeutic window between normal tissue complication and tumour control. Previously, MRT was limited to ultra-high dose rate synchrotron facilities. The aim of this study was to investigate in vitro effects of MRT on tumour and normal cells at conventional dose rates produced by a bench-top X-ray source. Two normal and two tumour cell lines were exposed to homogeneous broad beam (BB) radiation, MRT, or were separately irradiated with peak or valley doses before being mixed. Clonogenic survival was assessed and compared to BB-estimated surviving fractions calculated by the linear-quadratic (LQ)-model. All cell lines showed similar BB sensitivity. BB LQ-model predictions exceeded the survival of cell lines following MRT or mixed beam irradiation. This effect was stronger in tumour compared to normal cell lines. Dose mixing experiments could reproduce MRT survival. We observed a differential response of tumour and normal cells to spatially fractionated irradiations in vitro, indicating increased tumour cell sensitivity. Importantly, this was observed at dose rates precluding the presence of FLASH effects. The LQ-model did not predict cell survival when the cell population received split irradiation doses, indicating that factors other than local dose influenced survival after irradiation. AU - Steel, H.* AU - Brüningk, S.C.* AU - Box, C.* AU - Oelfke, U.* AU - Bartzsch, S. C1 - 62382 C2 - 50852 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Quantification of differential response of tumour and normal cells to microbeam radiation in the absence of flash effects. JO - Cancers VL - 13 IS - 13 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - Two contributors’ names were missing in the original version of the authorship of the paper [1]. The two authors are Markus Kaller and Heiko Hermeking, who should be listed as the sixth and seventh authors. Their affiliations are (Markus Kaller 5, Heiko Hermeking 5,6,7): 5 Experimental and Molecular Pathology, Institute of Pathology, Ludwig-Maximilians-University Munich, 80337 Munich, Germany; markus.kaller@med.uni-muenchen.de (M.K.); heiko.hermeking@med.uni-muenchen.de (H.H.) 6 German Cancer Consortium (DKTK), Partner Site Munich, 80336 Munich, Germany 7 German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany The “Author Contributions” statement thus should be updated to the following version. AU - Wagner, A.E.* AU - Schwarzmayr, T. AU - Häberle, B.* AU - Vokuhl, C.* AU - Schmid, I.* AU - Kaller, M.* AU - Hermeking, H.* AU - von Schweinitz, D.* AU - Kappler, R.* C1 - 61115 C2 - 49842 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Erratum: Wagner, A.E., et al. SP8 promotes an aggressive phenotype in hepatoblastoma via FGF8 activation. (Cancers 2020, 12, 2294). JO - Cancers VL - 13 IS - 3 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - Advanced radiation techniques can reduce the severity of neurocognitive sequelae in young brain tumor patients. In the present analysis, we sought to compare neurocognitive outcomes after proton irradiation with patients who underwent photon radiotherapy (RT) and surgery. Neurocogni-tive outcomes were evaluated in 103 pediatric brain tumor patients (proton RT n = 26, photon RT n = 30, surgery n = 47) before and after treatment. Comparison of neurocognitive outcomes following different treatment modalities were analyzed over four years after treatment completion. Longitudinal analyses included 42 months of follow-up after proton RT and 55 months after photon RT and surgery. Neurocognitive assessment included standardized tests examining seven domains. A comparison of neurocognitive outcomes after RT (proton and photon with >90% additional surgery) and surgery showed no significant differences in any neurocognitive domain. Neurocognitive functioning tests after proton RT failed to identify alterations compared to baseline testing. Long-term follow up over four years after photon RT showed a decrease in non-verbal intelligence (−9.6%; p = 0.01) and visuospatial construction (−14.9%; p = 0.02). After surgery, there was a decline in non-verbal intelligence (−10.7%; p = 0.01) and processing speed (14.9%; p = 0.002). Differences in neurocognitive outcomes between RT and surgical cohorts in direct intermodal comparison at long-term follow-up were not identified in our study, suggesting that modern radiation therapy does not affect cognition as much as in the past. There were no alterations in long-term neurocognitive abilities after proton RT, whereas decline of processing speed, non-verbal intelligence, and visuospatial abilities were observed after both photon RT and surgery. Domains dependent on intact white matter structures appear particularly vulnerable to brain tumor treatment irrespective of treatment approach. AU - Weusthof, K.* AU - Lüttich, P.* AU - Regnery, S.* AU - König, L.M.* AU - Bernhardt, D. AU - Witt, O.* AU - Unterberg, A.* AU - Jungk, C.* AU - Farnia, B.* AU - Combs, S.E. AU - Rieken, S.* C1 - 62618 C2 - 50913 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Neurocognitive outcomes in pediatric patients following brain irradiation. JO - Cancers VL - 13 IS - 14 PB - Mdpi PY - 2021 SN - 2072-6694 ER - TY - JOUR AB - The miR-221 expression is dependent on the oncogenic RAS-RAF-MEK pathway activation and influences epithelial-to-mesenchymal transition (EMT). The Cancer Genome Atlas (TCGA) database analysis showed high gene significance for ZEB1 with EMT module analysis and miR-221 overexpression within the triple-negative breast cancer (TNBC) and HER2+ subgroups when compared to luminal A/B subgroups. EMT marker expression analysis after MEK1 (TAK-733) inhibitor treatment and irradiation was combined with miR-221 and ZEB1 expression analysis. The interaction of miR-221 overexpression with irradiation and its influence on migration, proliferation, colony formation and subsequent EMT target activation were investigated. The results revealed that MEK1 inhibitor treatment combined with irradiation could decrease the migratory potential of breast cancer cells including reduction of miR-221 and corresponding downstream ZEB1 (EMT) marker expression. The clonogenic survival assays revealed that miR-221 overexpressing SKBR3 cells were more radioresistant when compared to the control. Remarkably, the effect of miR-221 overexpression on migration in highly proliferative and highly HER2-positive SKBR3 cells remained constant even upon 8 Gy irradiation. Further, in naturally miR-221-overexpressing MDA-MB-231 cells, the proliferation and migration significantly decrease after miR-221 knockdown. This leads to the assumption that radiation alone is not reducing migration capacity of miR-221-overexpressing cells and that additional factors play an important role in this context. The miR-221/ZEB1 activity is efficiently targeted upon MEK1 inhibitor (TAK-733) treatment and when combined with irradiation treatment, significant reduction in migration of breast cancer cells was shown. AU - Anastasov, N. AU - Hirmer, E. AU - Klenner, M. AU - Ott, J. AU - Falkenberg, N.* AU - Bao, X. AU - Mutschelknaus, L. AU - Mörtl, S. AU - Combs, S.E. AU - Atkinson, M.J. AU - Schmid, T.E. C1 - 60841 C2 - 49659 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Mek1 inhibitor combined with irradiation reduces migration of breast cancer cells including mir-221 and zeb1 emt marker expression. JO - Cancers VL - 12 IS - 12 PB - Mdpi PY - 2020 SN - 2072-6694 ER - TY - JOUR AB - Despite it being the most common incident of cancer among men, the pathophysiological mechanisms contributing to prostate cancer (PCa) are still poorly understood. Altered mitochondrial metabolism is postulated to play a role in the development of PCa. To determine the key metabolites (which included mitochondrial oncometabolites), benign prostatic and cancer tissues of patients with PCa were analyzed using capillary electrophoresis and liquid chromatography coupled with mass spectrometry. Gene expression was studied using real-time PCR. In PCa tissues, we found reduced levels of early tricarboxylic acid cycle metabolites, whereas the contents of urea cycle metabolites including aspartate, argininosuccinate, arginine, proline, and the oncometabolite fumarate were higher than that in benign controls. Fumarate content correlated positively with the gene expression of oncogenic HIF1 alpha and NF kappa B pathways, which were significantly higher in the PCa samples than in the benign controls. Furthermore, data from the TCGA database demonstrated that prostate cancer patients with activated NF kappa B pathway had a lower survival rate. In summary, our data showed that fumarate content was positively associated with carcinogenic genes. AU - Frankó, A. AU - Shao, Y.* AU - Heni, M. AU - Hennenlotter, J.* AU - Hoene, M.* AU - Hu, C.* AU - Liu, X.* AU - Zhao, X.* AU - Wang, Q.* AU - Birkenfeld, A.L. AU - Todenhöfer, T.* AU - Stenzl, A.* AU - Peter, A.* AU - Häring, H.-U. AU - Lehmann, R. AU - Xu, G.* AU - Lutz, S.Z.* C1 - 59653 C2 - 48931 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Human prostate cancer is characterized by an increase in urea cycle metabolites. JO - Cancers VL - 12 IS - 7 PB - Mdpi PY - 2020 SN - 2072-6694 ER - TY - JOUR AB - Whether cancer patients receiving immune checkpoint inhibitors (ICI) are at an increased risk of severe infection and mortality during the corona pandemic is a hotly debated topic that will continue to evolve. Here, we summarize and discuss current studies regarding COVID-19 and anti-cancer treatment with an emphasis on ICI. Importantly, several lines of evidence suggest that patients currently treated with ICI do not display an increased vulnerability to infection with SARS-CoV-2. Data regarding morbidity and mortality associated with COVID-19 in cancer patients receiving ICI are less clear and often conflicting. Although mostly based on experimental data, it is possible that ICI can promote the exacerbated immune response associated with adverse outcome in COVID-19 patients. On the other hand, mounting evidence suggests that ICI might even be useful in the treatment of viral infections by preventing or ameliorating T cell exhaustion. In this context, the right timing of treatment might be essential. Nevertheless, some cancer patients treated with ICI experience autoimmune-related side effects that require the use of immunosuppressive therapies, which in turn may promote a severe course of infection with SARS-CoV-2. Although there is clear evidence that withholding ICI will have more serious consequences, further studies are urgently needed in to better evaluate the effects of ICI in patients with COVID-19 and the use of ICI during the corona pandemic in general. AU - Gambichler, T.* AU - Reuther, J.* AU - Scheel, C. AU - Susok, L.* AU - Kern, P.* AU - Becker, J.C.* C1 - 60573 C2 - 49415 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Cancer and immune checkpoint inhibitor treatment in the era of SARS-CoV-2 infection. JO - Cancers VL - 12 IS - 11 PB - Mdpi PY - 2020 SN - 2072-6694 ER - TY - JOUR AB - The expanded use of second-generation antiandrogens revolutionized the treatment landscape of progressed prostate cancer. However, resistances to these novel drugs are already the next obstacle to be solved. Various previous studies depicted an involvement of the enzyme AKR1C3 in the process of castration resistance as well as in the resistance to 2nd generation antiandrogens like enzalutamide. In our study, we examined the potential of natural AKR1C3 inhibitors in various prostate cancer cell lines and a three-dimensional co-culture spheroid model consisting of cancer cells and cancer-associated fibroblasts (CAFs) mimicking enzalutamide resistant prostate cancer. One of our compounds, named MF-15, expressed strong antineoplastic effects especially in cell culture models with significant enzalutamide resistance. Furthermore, MF-15 exhibited a strong effect on androgen receptor (AR) signaling, including significant inhibition of AR activity, downregulation of androgen-regulated genes, lower prostate specific antigen (PSA) production, and decreased AR and AKR1C3 expression, indicating a bi-functional effect. Even more important, we demonstrated a persisting inhibition of AR activity in the presence of AR-V7 and further showed that MF-15 non-competitively binds within the DNA binding domain of the AR. The data suggest MF-15 as useful drug to overcome enzalutamide resistance. AU - Kafka, M.* AU - Mayr, F.* AU - Temml, V.* AU - Möller, G. AU - Adamski, J. AU - Höfer, J.* AU - Schwaiger, S.* AU - Heidegger, I.* AU - Matuszczak, B.* AU - Schuster, D.* AU - Klocker, H.* AU - Bektic, J.* AU - Stuppner, H.* AU - Eder, I.E.* C1 - 59828 C2 - 48245 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Dual inhibitory action of a novel AKR1C3 inhibitor on both full-length AR and the variant AR-V7 in enzalutamide resistant metastatic castration resistant prostate cancer. JO - Cancers VL - 12 IS - 8 PB - Mdpi PY - 2020 SN - 2072-6694 ER - TY - JOUR AB - Glioblastoma multiforme (GBM) is the most common high-grade intracranial tumor in adults. It is characterized by uncontrolled proliferation, diffuse infiltration due to high invasive and migratory capacities, as well as intense resistance to chemo- and radiotherapy. With a five-year survival of less than 3% and an average survival rate of 12 months after diagnosis, GBM has become a focus of current research to urgently develop new therapeutic approaches in order to prolong survival of GBM patients. The methylation status of the promoter region of the O-6-methylguanine-DNA methyltransferase (MGMT) is nowadays routinely analyzed since a methylated promoter region is beneficial for an effective response to temozolomide-based chemotherapy. Furthermore, several miRNAs were identified regulating MGMT expression, apart from promoter methylation, by degrading MGMT mRNA before protein translation. These miRNAs could be a promising innovative treatment approach to enhance Temozolomide (TMZ) sensitivity in MGMT unmethylated patients and to increase progression-free survival as well as long-term survival. In this review, the relevant miRNAs are systematically reviewed. AU - Kirstein, A. AU - Schmid, T.E. AU - Combs, S.E. C1 - 59006 C2 - 48523 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - The role of miRNA for the treatment of MGMT unmethylated glioblastoma multiforme. JO - Cancers VL - 12 IS - 5 PB - Mdpi PY - 2020 SN - 2072-6694 ER - TY - JOUR AB - Simple Summary: Radiomic risk models are usually based on imaging features, which are extracted from the entire gross tumour volume (GTVentire). This approach does not explicitly consider the complex biological structure of the tumours. Therefore, in this retrospective study, we investigated the prognostic value of radiomic analyses based on different tumour sub-volumes using computed tomography imaging of patients with locally advanced head and neck squamous cell carcinoma who were treated with primary radio-chemotherapy. The GTVentire was cropped by different margins to define the rim and corresponding core sub-volumes of the tumour. Furthermore, the best performing tumour rim sub-volume was extended into surrounding tissue with different margins. As a result, the models based on the 5 mm tumour rim and on the 3 mm extended rim sub-volume showed an improved performance compared to models based on the corresponding tumour core. This indicates that the consideration of tumour sub-volumes may help to improve radiomic risk models.Imaging features for radiomic analyses are commonly calculated from the entire gross tumour volume (GTVentire). However, tumours are biologically complex and the consideration of different tumour regions in radiomic models may lead to an improved outcome prediction. Therefore, we investigated the prognostic value of radiomic analyses based on different tumour sub-volumes using computed tomography imaging of patients with locally advanced head and neck squamous cell carcinoma. The GTVentire was cropped by different margins to define the rim and the corresponding core sub-volumes of the tumour. Subsequently, the best performing tumour rim sub-volume was extended into surrounding tissue with different margins. Radiomic risk models were developed and validated using a retrospective cohort consisting of 291 patients in one of the six Partner Sites of the German Cancer Consortium Radiation Oncology Group treated between 2005 and 2013. The validation concordance index (C-index) averaged over all applied learning algorithms and feature selection methods using the GTVentire achieved a moderate prognostic performance for loco-regional tumour control (C-index: 0.61 +/- 0.04 (mean +/- std)). The models based on the 5 mm tumour rim and on the 3 mm extended rim sub-volume showed higher median performances (C-index: 0.65 +/- 0.02 and 0.64 +/- 0.05, respectively), while models based on the corresponding tumour core volumes performed less (C-index: 0.59 +/- 0.01). The difference in C-index between the 5 mm tumour rim and the corresponding core volume showed a statistical trend (p = 0.10). After additional prospective validation, the consideration of tumour sub-volumes may be a promising way to improve prognostic radiomic risk models. AU - Leger, S.* AU - Zwanenburg, A.* AU - Leger, K.* AU - Lohaus, F.* AU - Linge, A.* AU - Schreiber, A.* AU - Kalinauskaite, G.* AU - Tinhofer, I.* AU - Guberina, N.* AU - Guberina, M.* AU - Balermpas, P.* AU - Von der Grün, J.* AU - Ganswindt, U. AU - Belka, C. AU - Peeken, J.C. AU - Combs, S.E. AU - Boeke, S.* AU - Zips, D.* AU - Richter, C.* AU - Krause, M.* AU - Baumann, M.* AU - Troost, E.G.C.* AU - Löck, S.* C1 - 60387 C2 - 49393 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Comprehensive analysis of tumour sub-volumes for radiomic risk modelling in locally advanced HNSCC. JO - Cancers VL - 12 IS - 10 PB - Mdpi PY - 2020 SN - 2072-6694 ER - TY - JOUR AB - Diffusion tensor imaging (DTI), and fractional-anisotropy (FA) maps in particular, have shown promise in predicting areas of tumor recurrence in glioblastoma. However, analysis of peritumoral edema, where most recurrences occur, is impeded by free-water contamination. In this study, we evaluated the benefits of a novel, deep-learning-based approach for the free-water correction (FWC) of DTI data for prediction of later recurrence. We investigated 35 glioblastoma cases from our prospective glioma cohort. A preoperative MR image and the first MR scan showing tumor recurrence were semiautomatically segmented into areas of contrast-enhancing tumor, edema, or recurrence of the tumor. The 10th, 50th and 90th percentiles and mean of FA and mean-diffusivity (MD) values (both for the original and FWC-DTI data) were collected for areas with and without recurrence in the peritumoral edema. We found significant differences in the FWC-FA maps between areas of recurrence-free edema and areas with later tumor recurrence, where differences in noncorrected FA maps were less pronounced. Consequently, a generalized mixed-effect model had a significantly higher area under the curve when using FWC-FA maps (AUC = 0.9) compared to noncorrected maps (AUC = 0.77, p < 0.001). This may reflect tumor infiltration that is not visible in conventional imaging, and may therefore reveal important information for personalized treatment decisions. AU - Metz, M.C.* AU - Molina-Romero, M.* AU - Lipkova, J.* AU - Gempt, J.* AU - Liesche-Starnecker, F.* AU - Eichinger, P.* AU - Grundl, L.* AU - Menze, B.* AU - Combs, S.E. AU - Zimmer, C.* AU - Wiestler, B.* C1 - 58684 C2 - 48295 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Predicting glioblastoma recurrence from preoperative MR scans using fractional-anisotropy maps with free-water suppression. JO - Cancers VL - 12 IS - 3 PB - Mdpi PY - 2020 SN - 2072-6694 ER - TY - JOUR AB - Today, pancreatic cancer is the seventh leading cause of cancer-related deaths worldwide with a five-year overall survival rate of less than 7%. Only 15–20% of patients are eligible for curative intent surgery at the time of diagnosis. Therefore, neoadjuvant treatment regimens have been introduced in order to downsize the tumor by chemotherapy and radiotherapy. To further increase the efficacy of radiotherapy, novel molecular biomarkers are urgently needed to define the subgroup of pancreatic cancer patients who would benefit most from radiotherapy. MicroRNAs (miRNAs) could have the potential to serve as novel predictive and prognostic biomarkers in patients with pancreatic cancer. In the present article, the role of miRNAs as blood biomarkers, which are associated with either radioresistance or radiation-induced changes of miRNAs in pancreatic cancer, is discussed. Furthermore, the manuscript provides own data of miRNAs identified in a pancreatic cancer mouse model as well as radiation-induced miRNA changes in the plasma of tumor-bearing mice. AU - Nguyen, L. AU - Schilling, D. AU - Dobiasch, S. AU - Raulefs, S. AU - Santiago Franco, M. AU - Buschmann, D.* AU - Pfaffl, M.W.* AU - Schmid, T.E. AU - Combs, S.E. C1 - 60897 C2 - 49658 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - The emerging role of miRNAs for the radiation treatment of pancreatic cancer. JO - Cancers VL - 12 IS - 12 PB - Mdpi PY - 2020 SN - 2072-6694 ER - TY - JOUR AB - Currently, decision-making regarding biochemical recurrence (BCR) following prostatectomy relies solely on clinical parameters. We therefore attempted to develop an integrated prediction model based on a molecular signature and clinicopathological features, in order to forecast the risk for BCR and guide clinical decision-making for postoperative therapy. Using highthroughput screening and least absolute shrinkage and selection operator (LASSO) in the training set, a novel gene signature for biochemical recurrence-free survival (BCRFS) was established. Validation of the prognostic value was performed in five other independent datasets, including our patient cohort. Multivariate Cox regression analysis was performed to evaluate the importance of risk for BCR. Time-dependent receiver operating characteristic (tROC) was used to evaluate the predictive power. In combination with relevant clinicopathological features, a decision tree was built to improve the risk stratification. The gene signature exhibited a strong capacity for identifying high-risk BCR patients, and multivariate Cox regression analysis demonstrated that the gene signature consistently acted as a risk factor for BCR. The decision tree was successfully able to identify the high-risk subgroup. Overall, the gene signature established in the present study is a powerful predictor and risk factor for BCR after radical prostatectomy. AU - Shi, R.* AU - Bao, X. AU - Weischenfeldt, J.* AU - Schaefer, C.* AU - Rogowski, P.* AU - Schmidt-Hegemann, N.S.* AU - Unger, K. AU - Lauber, K. AU - Wang, X.* AU - Buchner, A.* AU - Stief, C.* AU - Schlomm, T.* AU - Belka, C. AU - Li, M.* C1 - 57700 C2 - 47876 TI - A novel gene signature-based model predicts biochemical recurrence-free survival in prostate cancer patients after radical prostatectomy. JO - Cancers VL - 12 IS - 1 PY - 2020 SN - 2072-6694 ER - TY - JOUR AB - More accurate models are essential to identify high-risk bladder cancer (BCa) patients who will benefit from adjuvant therapies and thus helpful to facilitate personalized management of BCa. Among various cancer-related hallmarks and pathways, cell cycle process (CCP) was identified as a dominant risk factor for cancer-specific survival (CSS) in BCa. Using a series of bioinformatic and statistical approaches, a CCP-related gene signature was established, and the prognostic value was validated in other independent BCa cohorts. In addition, the risk score derived from the gene signature serves as a promising marker for therapeutic resistance. In combination with clinicopathological features, a nomogram was constructed to provide more accurate prediction for CSS, and a decision tree was built to identify high-risk subgroup of muscle invasive BCa patients. Overall, the gene signature could be a useful tool to predict CSS and help to identify high-risk subgroup of BCa patients, which may benefit from intensified adjuvant therapy. AU - Shi, R.* AU - Bao, X* AU - Rogowski, P.* AU - Schäfer, C.* AU - Schmidt-Hegemann, N.S.* AU - Unger, K. AU - Lu, S.* AU - Sun, J.* AU - Buchner, A.* AU - Stief, C.* AU - Belka, C.* AU - Li, M.* C1 - 59040 C2 - 48665 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Establishment and validation of an individualized cell cycle process-related gene signature to predict cancer-specific survival in patients with bladder cancer. JO - Cancers VL - 12 IS - 5 PB - Mdpi PY - 2020 SN - 2072-6694 ER - TY - JOUR AB - Hepatoblastoma (HB) is the most common malignant liver tumor in childhood and it generally has a good prognosis. However, if associated with aggressive metastatic disease, outcome is still poor. The molecular mechanisms leading to metastatic spread in HB patients are still unknown. By combining RNA-sequencing and a genome-wide methylome analysis, we identified the transcription factor SP8 and the growth factor FGF8 among the most strongly upregulated genes in metastatic HB cases, with a concomitant robust demethylation of the respective promoter regions. Of note, high expression of both candidates was associated with the aggressive C2 subtype of the 16-gene signature and poor survival. Chromatin immunoprecipitation revealed a direct transcriptional regulation of FGF8 through binding of SP8 to theFGF8promoter. Gain- and loss-of-function experiments proved promoting effects of SP8 on motility, self-renewal, migration, and the invasive potential of HB cells. Moreover, stable overexpression of SP8 in Hep3B cells resulted in the acquisition of a mesenchymal phenotype and a strong upregulation of epithelial-mesenchymal transition-associated genes. Using KRAB-mediated CRISPR-dCas9 interference directed against FGF8, we could show that FGF8 is essential for the SP8-mediated aggressive tumor behavior. Treatment of HB cell lines with the pan SP family inhibitor mithramycin A resulted in a significant inhibition of their clonogenic growth. In summary, we identified SP8 and FGF8 as key players in aggressive traits of HB and propose SP8 inhibiting drugs as a new effective treatment strategy especially for metastatic tumors. AU - Wagner, A.E.* AU - Schwarzmayr, T. AU - Häberle, B.* AU - Vokuhl, C.* AU - Schmid, I.* AU - von Schweinitz, D.* AU - Kappler, R.* C1 - 60021 C2 - 49171 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - SP8 promotes an aggressive phenotype in hepatoblastoma via FGF8 activation, JO - Cancers VL - 12 IS - 8 PB - Mdpi PY - 2020 SN - 2072-6694 ER - TY - JOUR AB - Pheochromocytomas and paragangliomas (PPGLs) with activated pseudohypoxic pathways are associated with an immature catecholamine phenotype and carry a higher risk for metastasis. For improved understanding of the underlying mechanisms we investigated the impact of hypoxia and pseudohypoxia on catecholamine biosynthesis in pheochromocytoma cells naturally lacking Hif2 alpha (MPC and MTT) or expressing both Hif1 alpha and Hif2 alpha (PC12). Cultivation under extrinsic hypoxia or in spheroid culture (intrinsic hypoxia) increased cellular dopamine and norepinephrine contents in all cell lines. To distinguish further between Hifla- and Hif2 alpha-driven effects we expressed Hif2 alpha in MTT and MPC-mCherry cells (naturally lacking Hif2 alpha). Presence of Hif2 alpha resulted in similarly increased cellular dopamine and norepinephrine under hypoxia as in the control cells. Furthermore, hypoxia resulted in enhanced phosphorylation of tyrosine hydroxylase (TH). A specific knockdown of Hif1 alpha in PC12 diminished these effects. Pseudohypoxic conditions, simulated by expression of Hif2 alpha under normoxia resulted in increased TH phosphorylation, further stimulated by extrinsic hypoxia. Correlations with PPGL tissue data led us to conclude that catecholamine biosynthesis under hypoxia is mainly mediated through increased phosphorylation of TH, regulated as a short-term response (24-48 h) by HIf1 alpha. Continuous activation of hypoxia-related genes under pseudohypoxia leads to a HIF2 alpha-mediated phosphorylation of TH (permanent status). AU - Bechmann, N.* AU - Poser, I.* AU - Seifert, V.* AU - Greunke, C. AU - Ullrich, M.* AU - Qin, N.* AU - Walch, A.K. AU - Peitzsch, M.* AU - Robledo, M.* AU - Pacak, K.* AU - Pietzsch, J.* AU - Richter, S.* AU - Eisenhofer, G.* C1 - 55961 C2 - 46723 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Impact of extrinsic and intrinsic hypoxia on catecholamine biosynthesis in absence or presence of Hif2 alpha in pheochromocytoma cells. JO - Cancers VL - 11 IS - 5 PB - Mdpi PY - 2019 SN - 2072-6694 ER - TY - JOUR AB - The use of different scoring systems for radiation-induced toxicity limits comparability between studies. We examined dose-dependent tissue alterations following hypofractionated X-ray irradiation and evaluated their use as scoring criteria. Four dose fractions (0, 5, 10, 20, 30 Gy/fraction) were applied daily to ear pinnae. Acute effects (ear thickness, erythema, desquamation) were monitored for 92 days after fraction 1. Late effects (chronic inflammation, fibrosis) and the presence of transforming growth factor beta 1 (TGF beta 1)-expressing cells were quantified on day 92. The maximum ear thickness displayed a significant positive correlation with fractional dose. Increased ear thickness and erythema occurred simultaneously, followed by desquamation from day 10 onwards. A significant dose-dependency was observed for the severity of erythema, but not for desquamation. After 4 x 20 and 4 x 30 Gy, inflammation was significantly increased on day 92, whereas fibrosis and the abundance of TGF beta 1-expressing cells were only marginally increased after 4 x 30 Gy. Ear thickness significantly correlated with the severity of inflammation and fibrosis on day 92, but not with the number of TGF beta 1-expressing cells. Fibrosis correlated significantly with inflammation and fractional dose. In conclusion, the parameter of ear thickness can be used as an objective, numerical and dose-dependent quantification criterion to characterize the severity of acute toxicity and allow for the prediction of late effects. AU - Dombrowsky, A. AU - Schauer, J.* AU - Sammer, M.* AU - Blutke, A. AU - Walsh, D.W.M.* AU - Schwarz, B.* AU - Bartzsch, S. AU - Feuchtinger, A. AU - Reindl, J.* AU - Combs, S.E. AU - Dollinger, G.* AU - Schmid, T.E. C1 - 56158 C2 - 46855 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Acute skin damage and late radiation-induced fibrosis and inflammation in murine ears after high-dose irradiation. JO - Cancers VL - 11 IS - 5 PB - Mdpi PY - 2019 SN - 2072-6694 ER - TY - JOUR AB - About 95% of patients with Glioblastoma (GBM) show tumor relapse, leaving them with limited therapeutic options as recurrent tumors are most often resistant to the first line chemotherapy standard Temozolomide (TMZ). To identify molecular pathways involved in TMZ resistance, primary GBM Stem-like Cells (GSCs) were isolated, characterized, and selected for TMZ resistance in vitro. Subsequently, RNA sequencing analysis was performed and revealed a total of 49 differentially expressed genes (|log2-fold change| > 0.5 and adjusted p-value < 0.1) in TMZ resistant stem-like cells compared to their matched DMSO control cells. Among up-regulated genes, we identified carbonic anhydrase 2 (CA2) as a candidate gene correlated with glioma malignancy and patient survival. Notably, we describe consistent up-regulation of CA2 not only in TMZ resistant GSCs on mRNA and protein level, but also in patient-matched clinical samples of first manifest and recurrent tumors. Co-treatment with the carbonic anhydrase inhibitor Acetazolamid (ACZ) sensitized cells to TMZ induced cell death. Cumulatively, our findings illustrate the potential of CA2 as a chemosensitizing target in recurrent GBM and provide a rationale for a therapy associated inhibition of CA2 to overcome TMZ induced chemoresistance. AU - Hannen, R.* AU - Selmansberger, M. AU - Hauswald, M.* AU - Pagenstecher, A.* AU - Nist, A.* AU - Stiewe, T.* AU - Acker, T.* AU - Carl, B.* AU - Nimsky, C.* AU - Bartsch, J.W.* C1 - 56563 C2 - 47141 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Comparative transcriptomic analysis of temozolomide resistant primary GBM stem-like cells and recurrent GBM identifies up-regulation of the carbonic anhydrase CA2 gene as resistance factor. JO - Cancers VL - 11 IS - 7 PB - Mdpi PY - 2019 SN - 2072-6694 ER - TY - JOUR AB - High-precision radiotherapy has been established as a valid and effective treatment option in patients with pituitary adenomas. We report on outcome after fractionated stereotactic radiotherapy (FSRT) in correlation with patient-reported outcomes (PROs). We analyzed 69 patients treated between 2000 and 2019. FSRT was delivered with a median total dose of 54 Gy (single fraction: 1.8 Gy). PRO questionnaires were sent to 28 patients. Median overall survival was 17.2 years; mean local control was 15.6 years (median not reached). Median follow-up was 5.8 years. Twenty (71%) patients participated in the PRO assessment. Physicians reported symptoms grade >= 3 in 6 cases (9%). Of all, 35 (51%) patients suffered from hypopituitarism at baseline, and during follow-up, new or progressive hypopituitarism was observed in 11 cases (16%). Patients reported 10 cases of severe side effects. Most of these symptoms were already graded as CTCAE (Common Terminology Criteria for Adverse Events) grade 2 by a physician in a previous follow-up exam. PROs are an essential measure and only correlate to a certain extent with the physician-reported outcomes. For high-precision radiotherapy of pituitary adenomas, they confirm excellent overall outcomes and low toxicity. In the future, the integration of PROs paired with high-end treatment will further improve outcomes. AU - Kessel, K.A. AU - Diehl, C.D.* AU - Oechsner, M.* AU - Meyer, B.* AU - Gempt, J.* AU - Zimmer, C.* AU - Schmidt-Graf, F.* AU - Combs, S.E. C1 - 57469 C2 - 47809 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Patient-reported outcome (PRO) as an addition to long-term results after high-precision stereotactic radiotherapy in patients with secreting and non-secreting pituitary adenomas: A retrospective cohort study up to 17-years follow-up. JO - Cancers VL - 11 IS - 12 PB - Mdpi PY - 2019 SN - 2072-6694 ER - TY - JOUR AB - The I-kappa B-Kinase (IKK) complex represents a central signaling nexus in the TNF-dependent activation of the pro-inflammatory NF-kappa B pathway. However, recent studies suggested that the distinct IKK subunits (IKK alpha, IKK beta, and NEMO) might withhold additional NF-kappa B-independent functions in inflammation and cancer. Here, we generated mice lacking all three IKK subunits in liver parenchymal cells (LPC) (IKK alpha/beta/NEMOLPC-KO) and compared their phenotype with mice lacking both catalytic subunits (IKK alpha/beta(LPC-KO)), allowing to functionally dissect putative I-kappa B-Kinase-independent functions of the regulatory subunit NEMO. We show that the additional deletion of NEMO rescues IKK alpha/beta(LPC-KO) mice from lethal cholestasis and biliary ductopenia by triggering LPC apoptosis and inducing a strong compensatory proliferation of LPC including cholangiocytes. Beyond this beneficial effect, we show that increased hepatocyte cell-death and compensatory proliferation inhibit the activation of LPC-necroptosis but trigger spontaneous hepatocarcinogenesis in IKK alpha/beta/NEMOLPC-KO mice. Collectively, our data show that free NEMO molecules unbound to the catalytic IKK subunits control LPC programmed cell death pathways and proliferation, cholestasis and hepatocarcinogenesis independently of an IKK-related function. These findings support the idea of different functional levels at which NEMO controls inflammation and cancer in the liver. AU - Koppe, C.* AU - Reisinger, F. AU - Wehr, K.* AU - Vucur, M.* AU - Trautwein, C.* AU - Tacke, F.* AU - Heikenwalder, M.* AU - Luedde, T.* C1 - 56599 C2 - 47165 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - An NF-kappaB- and IKK-independent function of NEMO prevents hepatocarcinogenesis by suppressing compensatory liver regeneration. JO - Cancers VL - 11 IS - 7 PB - Mdpi PY - 2019 SN - 2072-6694 ER - TY - JOUR AB - Background: Surgical resection offers the best chance of survival in patients with pancreatic cancer, but those with locally advanced disease (LAPC) are usually not surgical candidates. This cohort often receives either neoadjuvant chemotherapy or chemoradiation (CRT), but unintended weight loss coupled with muscle wasting (sarcopenia) can often be observed. Here, we report on the predictive value of changes in weight and muscle mass in 147 consecutive patients with LAPC treated with neoadjuvant CRT. Methods: Clinicopathologic data were obtained via a retrospective chart review. The abdominal skeletal muscle area (SMA) at the third lumbar vertebral body was determined via computer tomographic (CT) scans as a surrogate for the muscle mass and skeletal muscle index (SMI) calculated. Uni- and multi-variable statistical tests were performed to assess for impact on survival. Results: Weight loss (14.5 vs. 20.3 months; p = 0.04) and loss of muscle mass (15.1 vs. 22.2 months; p = 0.007) were associated with poor outcomes. The highest survival was observed in patients who had neither cachectic weight loss nor sarcopenia (27 months), with improved survival seen in those who ultimately received a resection (23 vs. 10 months; p < 0.001). Cox regression revealed that either continued weight loss or continued muscle wasting (SMA reduction) was predictive of poor outcomes, whereas a sarcopenic SMI was not. Conclusions: Loss of weight and lean muscle in patients with LAPC is prognostic when persistent. Therefore, both should be assessed longitudinally and considered before surgery. AU - Naumann, P.* AU - Eberlein, J.* AU - Farnia, B.* AU - Hackert, T.* AU - Debus, J.* AU - Combs, S.E. C1 - 56165 C2 - 46868 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Continued weight loss and sarcopenia Ppedict poor outcomes in locally advanced pancreatic cancer treated with chemoradiation. JO - Cancers VL - 11 IS - 5 PB - Mdpi PY - 2019 SN - 2072-6694 ER - TY - JOUR AB - Background: Patients with pancreatic cancer often develop cancer cachexia, a complex multifactorial syndrome with weight loss, muscle wasting and adipose tissue depletion with systemic inflammation causing physical impairment. In patients with locally advanced pancreatic cancer (LAPC) neoadjuvant treatment is routinely performed to allow a subsequent resection. Herein, we assess body composition and laboratory markers for cancer cachexia both before and after neoadjuvant chemoradiation (CRT). Methods: Subcutaneous fat (SCF), visceral fat (VF), skeletal muscle (SM), weight and laboratory parameters were determined longitudinally in 141 LAPC patients treated with neoadjuvant CRT. Changes during CRT were statistically analyzed and correlated with outcome and Kaplan-Meier curves were plotted. Different prognostic factors linked to cachexia were assessed by uni- and multivariable cox proportional hazards models. Results: There was a significant decrease in weight as well as SCF, VF and SM during CRT. The laboratory parameter C-reactive protein (CRP) increased significantly, whereas there was a significant decrease in leukocyte count, hemoglobin, albumin and cholinesterase as well as in the tumor marker CA 19.9. Cachectic weight loss, sarcopenia, reductions in body compartments SCF, VF and SM, and changes in laboratory markers as well as resection affected survival in univariable analysis. In multivariable analysis, weight loss >5% (HR 2.8), reduction in SM >5% (HR 5.5), an increase in CRP (HR 2.2) or CA 19.9 (HR 1.9), and resection (HR 0.4) remained independently associated with survival, whereas classical cachexia and sarcopenia did not. Interestingly, the subgroup of patients with cachectic weight loss >5% or SM reduction >5% during CRT did not benefit from resection (median survival 12 vs. 27 months). Conclusions: Persistent weight loss and muscle depletion during CRT as well as systemic inflammation after CRT impacted survival more than cachexia or sarcopenia according classical definitions. AU - Naumann, P.* AU - Eberlein, J.* AU - Farnia, B.* AU - Liermann, J.* AU - Hackert, T.* AU - Debus, J.* AU - Combs, S.E. C1 - 57285 C2 - 47712 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Cachectic body composition and inflammatory markers portend a poor prognosis in patients with locally advanced pancreatic cancer treated with chemoradiation. JO - Cancers VL - 11 IS - 11 PB - Mdpi PY - 2019 SN - 2072-6694 ER - TY - JOUR AB - (1) Background: Selecting patients that will benefit the most from proton radiotherapy (PRT) is of major importance. This study sought to assess dose reductions to numerous organs-at-risk (OARs) with PRT, as compared to three-dimensional conformal radiotherapy (3DCRT) and volumetric-modulated arc therapy (VMAT), as a function of tumor location. (2) Materials/Methods: Patients with intracranial neoplasms (all treated with PRT) were stratified into five location-based groups (frontal, suprasellar, temporal, parietal, posterior cranial fossa; n = 10 per group). Each patient was re-planned for 3DCRT and intensity-modulated radiotherapy (IMRT) using similar methodology, including the originally planned target and organ-at-risk (OAR) dose constraints. (3) Results: In parietal tumors, PRT showed the most pronounced dose reductions. PRT lowered doses to nearly every OAR, most notably the optical system and several contralateral structures (subventricular zone, thalamus, hippocampus). For frontal lobe cases, the greatest relative dose reductions in mean dose (D-mean) with PRT were to the infratentorial normal brain, contralateral hippocampus, brainstem, pituitary gland and contralateral optic nerve. For suprasellar lesions, PRT afforded the greatest relative D-mean reductions to the infratentorial brain, supratentorial brain, and the whole brain. Similar results could be observed in temporal and posterior cranial fossa disease. (4) Conclusions: The effectiveness and degree of PRT dose-sparing to various OARs depends on intracranial tumor location. These data will help to refine selection of patients receiving PRT, cost-effectiveness, and future clinical toxicity assessment. AU - Adeberg, S.* AU - Harrabi, S.B.* AU - Bougatf, N.* AU - Verma, V.* AU - Windisch, P.* AU - Bernhardt, D.* AU - Combs, S.E. AU - Herfarth, K.* AU - Debus, J.* AU - Rieken, S.* C1 - 54736 C2 - 45826 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Dosimetric comparison of proton radiation therapy, volumetric modulated arc therapy, and three-dimension conformal radiotherapy based on intracranial tumor location. JO - Cancers VL - 10 IS - 11 PB - Mdpi PY - 2018 SN - 2072-6694 ER - TY - JOUR AB - Gliomas are primary brain tumors that present the majority of malignant adult brain tumors. Gliomas are subdivided into low- and high-grade tumors. Despite extensive research in recent years, the prognosis of malignant glioma patients remains poor. This is caused by naturally highly infiltrative capacities as well as high levels of radio- and chemoresistance. Additionally, it was shown that low linear energy transfer (LET) irradiation enhances migration and invasion of several glioma entities which might counteract today's treatment concepts. However, this finding is discussed controversially. In the era of personalized medicine, this controversial data might be attributed to the patient-specific heterogeneity that ultimately could be used for treatment. Thus, current developments in glioma therapy should be seen in the context of intrinsic and radiation-enhanced migration and invasion. Due to the natural heterogeneity of glioma cells and different radiation responses, a personalized radiation treatment concept is suggested and alternative radiation concepts are discussed. AU - Wank, M. AU - Schilling, D. AU - Schmid, T.E. AU - Meyer, B.* AU - Gempt, J.* AU - Barz, M.* AU - Schlegel, J.* AU - Liesche, F.* AU - Kessel, K.A. AU - Wiestler, B.* AU - Bette, S.* AU - Zimmer, C.* AU - Combs, S.E. C1 - 54859 C2 - 45902 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Human glioma migration and infiltration properties as a target for personalized radiation medicine. JO - Cancers VL - 10 IS - 11 PB - Mdpi PY - 2018 SN - 2072-6694 ER - TY - JOUR AB - Glioblastoma (GBM) is the most common and most aggressive malignant primary brain tumor in adults. In spite of multimodal therapy concepts, consisting of surgery, radiotherapy and chemotherapy, the median survival, merely 15–18 months, is still poor. Mechanisms for resistance of GBM to radio(chemo)therapy are not fully understood yet and due to the genetic heterogeneity within the tumor including radiation-resistant tumor stem cells, there are several factors leading to therapy failure. Recent research revealed that, hypoxia during radiation and miRNAs may adversely affect the therapeutic response to radiotherapy. Further molecular alterations and prognostic markers like the DNA-repair protein O6-methylguanine-DNA methyltransferase (MGMT), anti-apoptotic molecular chaperones, and/or the activity of aldehyde dehydrogenase 1 (ALDH1) have also been identified to play a role in the sensitivity to cytostatic agents. Latest approaches in the field of radiotherapy to use particle irradiation or dose escalation strategies including modern molecular imaging, however, need further evaluation with regard to long-term outcome. In this review we focus on current information about the mechanisms and markers that mediate resistance to radio(chemo)therapy, and discuss the opportunities of Innovative Radiotherapy (iRT) concepts to improve treatment options for GBM patients. AU - Combs, S.E. AU - Schmid, T.E. AU - Vaupel, P.* AU - Multhoff, G. C1 - 47781 C2 - 39530 TI - Stress response leading to resistance in glioblastoma: The need for innovative radiotherapy (iRT) concepts. JO - Cancers VL - 8 IS - 1 PY - 2016 SN - 2072-6694 ER - TY - JOUR AB - Newly formed microvessels in most solid tumors show an abnormal morphology and thus do not fulfil the metabolic demands of the growing tumor mass. Due to the chaotic and heterogeneous tumor microcirculation, a hostile tumor microenvironment develops, that is characterized inter alia by local hypoxia, which in turn can stimulate the HIF-system. The latter can lead to tumor progression and may be involved in hypoxia-mediated radioresistance of tumor cells. Herein, cellular and molecular mechanisms in tumor angiogenesis are discussed that, among others, might impact hypoxia-related radioresistance. AU - Multhoff, G. AU - Radons, J.R.* AU - Vaupel, P.W.* C1 - 31591 C2 - 34575 SP - 813-823 TI - Critical role of aberrant angiogenesis in the development of tumor hypoxia and associated radioresistance. JO - Cancers VL - 6 IS - 2 PY - 2014 SN - 2072-6694 ER - TY - JOUR AB - Her2 overexpression and amplification can be found in a significant subset of esophageal adenocarcinomas. The activity of Her2 has been shown to be modulated by molecular chaperones such as HSP90. We analyzed expression/amplification data for HSP90 and Her2 on 127 primary resected esophageal adenocarcinomas in order to evaluate a possible relationship between these two molecules. HSP90 expression determined by immunohistochemistry was observed in various levels. Thirty nine (39) tumors (30.7%) were classified as Her2-positive according to their immunoreactivity and amplification status. There was a significant correlation between HSP90 expression and Her2-status (p = 0.008). This could also be demonstrated by quantitative protein expression analysis with reverse phase protein arrays (r = 0.9; p < 0.001). Her2-status was associated withpT-category (p = 0.041), lymph node metastases (p = 0.049) and tumor differentiation (p = 0.036) with a higher percentage of cases with negative Her2 status in lower tumor stagesA negative Her2-status was also associated with better survival in univariate and multivariate analysis (p = 0.001 and p = 0.014). For HSP90, no associations between clinical and pathological parameters were found. The observed association between HSP90 expression and Her2 suggests a co-regulation of these molecules in at least a subset of esophageal adenocarcinomas. Anti-HSP90 drugs, which recently have been introduced in cancer treatment, may also be an option for these tumors by targeting HSP90 alone or in combination with Her2. AU - Slotta-Huspenina, J.* AU - Becker, K.F.* AU - Feith, M.* AU - Walch, A.K. AU - Langer, R.* C1 - 31712 C2 - 34684 SP - 1382-1393 TI - Heat Shock Protein 90 (HSP90) and Her2 in adenocarcinomas of the esophagus. JO - Cancers VL - 6 IS - 3 PY - 2014 SN - 2072-6694 ER -