TY  - JOUR
AB  - BACKGROUND: Recurrent/metastatic head and neck squamous cell carcinoma (R/M-HNSCC) is a severe, frequently lethal condition. Oncogene addiction to epidermal growth factor receptor (EGFR) is a hallmark of HNSCC, but the clinical efficacy of EGFR-targeted therapies remains low. Understanding molecular networks governing EGFR-driven progression is paramount to the exploration of (co)-treatment targets and predictive markers. METHODS: We performed function-based mapping of differentially expressed genes in EGFR-mediated local invasion (fDEGs) using photoconvertible tracers and RNA-sequencing (RNA-seq) in a cellular 3D-model. RESULTS: Upon alignment with public single-cell RNA-seq (scRNA-seq) datasets and HNSCC-specific regulons, a gene regulatory network of local invasion (invGRN) was inferred from gene expression data, which was overrepresented in budding tumors. InvGRN comprises the central hubs inhibin subunit beta alpha (INHBA) and snail family transcriptional repressor 2 (SNAI2), and druggable fDEGs integrin subunit beta 4 (ITGB4), laminin 5 (LAMB3/LAMC2), and sphingosine kinase 1 (SPHK1). Blockade of INHBA repressed local invasion and was reverted by activin A, laminin 5, and sphingosine-1-phosphate, demonstrating a functional interconnectivity of the invGRN. Epithelial-to-mesenchymal transition (EMT) of malignant cells and the invGRN are induced by newly defined EGFR-activity subtypes with prognostic value that are promoted by amphiregulin (AREG) and epiregulin (EREG). Importantly, co-inhibition of SPHK1 showed synthetic effects on Cetuximab-mediated invasion blockade and high expression of selected fDEGs was associated with response to Cetuximab in patient-derived xenotransplantation (PDX) and R/M-HNSCC patients. CONCLUSIONS: We describe an actionable network of EGFR-mediated local invasion and define druggable effectors with predictive potential regarding the response of R/M-HNSCC to Cetuximab.
AU  - Zhou, J.*
AU  - He, M.*
AU  - Zhao, Q.*
AU  - Shi, E.*
AU  - Wang, H.*
AU  - Ponkshe, V.*
AU  - Song, J.*
AU  - Wu, Z.*
AU  - Ji, D.*
AU  - Kranz, G.*
AU  - Tscherne, A.*
AU  - Schwenk-Zieger, S.*
AU  - Razak, N.A.*
AU  - Hess J.
AU  - Belka, C.*
AU  - Zitzelsberger, H.
AU  - Ourailidis, I.*
AU  - Stögbauer, F.*
AU  - Boxberg, M.*
AU  - Budczies, J.*
AU  - Reichel, C.A.*
AU  - Canis, M.*
AU  - Baumeister, P.*
AU  - Unger, K.
AU  - Mock, A.*
AU  - Gires, O.*
C1  - 73736
C2  - 57203
CY  - Campus, 4 Crinan St, London N1 9xw, England
TI  - EGFR-mediated local invasiveness and response to Cetuximab in head and neck cancer.
JO  - Mol. Cancer
VL  - 24
IS  - 1
PB  - Bmc
PY  - 2025
ER  - 

TY  - JOUR
AB  - BACKGROUND: Acute leukemias represent deadly malignancies that require better treatment. As a challenge, treatment is counteracted by a microenvironment protecting dormant leukemia stem cells. METHODS: To identify responsible surface proteins, we performed deep proteome profiling on minute numbers of dormant patient-derived xenograft (PDX) leukemia stem cells isolated from mice. Candidates were functionally screened by establishing a comprehensive CRISPR‒Cas9 pipeline in PDX models in vivo. RESULTS: A disintegrin and metalloproteinase domain-containing protein 10 (ADAM10) was identified as an essential vulnerability required for the survival and growth of different types of acute leukemias in vivo, and reconstitution assays in PDX models confirmed the relevance of its sheddase activity. Of translational importance, molecular or pharmacological targeting of ADAM10 reduced PDX leukemia burden, cell homing to the murine bone marrow and stem cell frequency, and increased leukemia response to conventional chemotherapy in vivo. CONCLUSIONS: These findings identify ADAM10 as an attractive therapeutic target for the future treatment of acute leukemias.
AU  - Bahrami, E.
AU  - Schmid, J.P.
AU  - Jurinovic, V.
AU  - Becker, M.
AU  - Wirth, A.-K.
AU  - Ludwig, R.
AU  - Kreissig, S.*
AU  - Duque Angel, T.V.
AU  - Amend, D.
AU  - Hunt, K.
AU  - Öllinger, R.*
AU  - Rad, R.*
AU  - Frenz, J.M.*
AU  - Solovey, M.
AU  - Ziemann, F.*
AU  - Mann, M.*
AU  - Vick, B.
AU  - Wichmann, C.*
AU  - Herold, T.
AU  - Jayavelu, A.K.*
AU  - Jeremias, I.
C1  - 68136
C2  - 54614
CY  - Campus, 4 Crinan St, London N1 9xw, England
TI  - Combined proteomics and CRISPR‒Cas9 screens in PDX identify ADAM10 as essential for leukemia in vivo.
JO  - Mol. Cancer
VL  - 22
IS  - 1
PB  - Bmc
PY  - 2023
ER  - 

TY  - JOUR
AB  - BACKGROUND: Epidermal growth factor receptor (EGFR) is both a driver oncogene and a therapeutic target in advanced head and neck squamous cell carcinoma (HNSCC). However, response to EGFR treatment is inconsistent and lacks markers for treatment prediction. This study investigated EGFR-induced epithelial-to-mesenchymal transition (EMT) as a central parameter in tumor progression and identified novel prognostic and therapeutic targets, and a candidate predictive marker for EGFR therapy response. METHODS: Transcriptomic profiles were analyzed by RNA sequencing (RNA-seq) following EGFR-mediated EMT in responsive human HNSCC cell lines. Exclusive genes were extracted via differentially expressed genes (DEGs) and a risk score was determined through forward feature selection and Cox regression models in HNSCC cohorts. Functional characterization of selected prognostic genes was conducted in 2D and 3D cellular models, and findings were validated by immunohistochemistry in primary HNSCC. RESULTS: An EGFR-mediated EMT gene signature composed of n = 171 genes was identified in responsive cell lines and transferred to the TCGA-HNSCC cohort. A 5-gene risk score comprising DDIT4, FADD, ITGB4, NCEH1, and TIMP1 prognosticated overall survival (OS) in TCGA and was confirmed in independent HNSCC cohorts. The EGFR-mediated EMT signature was distinct from EMT hallmark and partial EMT (pEMT) meta-programs with a differing enrichment pattern in single malignant cells. Molecular characterization showed that ITGB4 was upregulated in primary tumors and metastases compared to normal mucosa and correlated with EGFR/MAPK activity in tumor bulk and single malignant cells. Preferential localization of ITGB4 together with its ligand laminin 5 at tumor-stroma interfaces correlated with increased tumor budding in primary HNSCC tissue sections. In vitro, ITGB4 knock-down reduced EGFR-mediated migration and invasion and ITGB4-antagonizing antibody ASC8 impaired 2D and 3D invasion. Furthermore, a logistic regression model defined ITGB4 as a predictive marker of progression-free survival in response to Cetuximab in recurrent metastatic HNSCC patients. CONCLUSIONS: EGFR-mediated EMT conveyed through MAPK activation contributes to HNSCC progression upon induction of migration and invasion. A 5-gene risk score based on a novel EGFR-mediated EMT signature prognosticated survival of HNSCC patients and determined ITGB4 as potential therapeutic and predictive target in patients with strong EGFR-mediated EMT.
AU  - Schinke, H.*
AU  - Shi, E.*
AU  - Lin, Z.*
AU  - Quadt, T.*
AU  - Kranz, G.*
AU  - Zhou, J.*
AU  - Wang, H.*
AU  - Hess J.
AU  - Heuer, S.
AU  - Belka, C.
AU  - Zitzelsberger, H.
AU  - Schumacher, U.*
AU  - Genduso, S.*
AU  - Riecken, K.*
AU  - Gao, Y.*
AU  - Wu, Z.*
AU  - Reichel, C.A.*
AU  - Walz, C.*
AU  - Canis, M.*
AU  - Unger, K.
AU  - Baumeister, P.
AU  - Pan, M.*
AU  - Gires, O.
C1  - 66150
C2  - 53091
TI  - A transcriptomic map of EGFR-induced epithelial-to-mesenchymal transition identifies prognostic and therapeutic targets for head and neck cancer.
JO  - Mol. Cancer
VL  - 21
IS  - 1
PY  - 2022
ER  - 

TY  - JOUR
AB  - BACKGROUND: Previous studies identified microRNAs (miRNAs) and messenger RNAs with significantly different expression between normal pancreas and pancreatic cancer (PDAC) tissues. Due to technological limitations of microarrays and real-time PCR systems these studies focused on a fixed set of targets. Expression of other RNA classes such as long intergenic non-coding RNAs or sno-derived RNAs has rarely been examined in pancreatic cancer. Here, we analysed the coding and non-coding transcriptome of six PDAC and five control tissues using next-generation sequencing. RESULTS: Besides the confirmation of several deregulated mRNAs and miRNAs, miRNAs without previous implication in PDAC were detected: miR-802, miR-2114 or miR-561. SnoRNA-derived RNAs (e.g. sno-HBII-296B) and piR-017061, a piwi-interacting RNA, were found to be differentially expressed between PDAC and control tissues. In silico target analysis of miR-802 revealed potential binding sites in the 3' UTR of TCF4, encoding a transcription factor that controls Wnt-signalling genes. Overexpression of miR-802 in MiaPaCa pancreatic cancer cells reduced TCF4 protein levels. Using Massive Analysis of cDNA Ends (MACE) we identified differential expression of 43 lincRNAs, long intergenic non-coding RNAs, e.g. LINC00261 and LINC00152 as well as several natural antisense transcripts like HNF1A-AS1 and AFAP1-AS1. Differential expression was confirmed by qPCR on the mRNA/miRNA/lincRNA level and by immunhistochemistry on the protein level. CONCLUSIONS: Here, we report a novel lncRNA, sncRNA and mRNA signature of PDAC. In silico prediction of ncRNA targets allowed for assigning potential functions to differentially regulated RNAs.
AU  - Müller, S.*
AU  - Raulefs, S.*
AU  - Bruns, P.*
AU  - Afonso-Grunz, F.*
AU  - Plötner, A.*
AU  - Thermann, R.*
AU  - Jager, C.*
AU  - Schlitter, A.M.*
AU  - Kong, B.*
AU  - Regel, I.*
AU  - Roth, W.K.*
AU  - Rotter, B.*
AU  - Hoffmeier, K.*
AU  - Kahl, G.F.*
AU  - Koch, I.*
AU  - Theis, F.J.
AU  - Kleeff, J.*
AU  - Winter, P.*
AU  - Michalski, C.W.*
C1  - 44480
C2  - 36947
CY  - London
TI  - Next-generation sequencing reveals novel differentially regulated mRNAs, lncRNAs, miRNAs, sdRNAs and a piRNA in pancreatic cancer.
JO  - Mol. Cancer
VL  - 14
IS  - 1
PB  - Biomed Central Ltd
PY  - 2015
ER  - 

TY  - JOUR
AU  - Müller, S.*
AU  - Raulefs, S.*
AU  - Bruns, P.*
AU  - Afonso-Grunz, F.*
AU  - Plötner, A.*
AU  - Thermann, R.*
AU  - Jager, C.*
AU  - Schlitter, A.M.*
AU  - Kong, B.*
AU  - Regel, I.*
AU  - Roth, W.K.*
AU  - Rotter, B.*
AU  - Hoffmeier, K.*
AU  - Kahl, G.F.*
AU  - Koch, I.*
AU  - Theis, F.J.
AU  - Kleeff, J.*
AU  - Winter, P.*
AU  - Michalski, C.W.*
C1  - 46500
C2  - 37616
CY  - London
TI  - Erratum to: Next-generation sequencing reveals novel differentially regulated mRNAs, lncRNAs, miRNAs, sdRNAs and a piRNA in pancreatic cancer.
JO  - Mol. Cancer
VL  - 14
IS  - 1
PB  - Biomed Central Ltd
PY  - 2015
ER  - 

TY  - JOUR
AB  - BACKGROUND: Syndromic forms of osteosarcoma (OS) account for less than 10% of all recorded cases of this malignancy. An individual OS predisposition is also possible by the inheritance of low penetrance alleles of tumor susceptibility genes, usually without evidence of a syndromic condition. Genetic variants involved in such a non-syndromic form of tumor predisposition are difficult to identify, given the low incidence of osteosarcoma cases and the genetic heterogeneity of patients. We recently mapped a major OS susceptibility QTL to mouse chromosome 14 by comparing alpha-radiation induced osteosarcoma in mouse strains which differ in their tumor susceptibility. METHODS: Tumor-specific allelic losses in murine osteosacoma were mapped along chromosome 14 using microsatellite markers and SNP allelotyping. Candidate gene search in the mapped interval was refined using PosMed data mining and mRNA expression analysis in normal osteoblasts. A strain-specific promoter variant in Rb1 was tested for its influence on mRNA expression using reporter assay. RESULTS: A common Rb1 allele derived from the BALB/cHeNhg strain was identified as the major determinant of radiation-induced OS risk at this locus. Increased OS-risk is linked with a hexanucleotide deletion in the promoter region which is predicted to change WT1 and SP1 transcription factor-binding sites. Both in-vitro reporter and in-vivo expression assays confirmed an approx. 1.5 fold reduced gene expression by this promoter variant. Concordantly, the 50% reduction in Rb1 expression in mice bearing a conditional hemizygous Rb1 deletion causes a significant rise of OS incidence following alpha-irradiation. CONCLUSION: This is the first experimental demonstration of a functional and genetic link between reduced Rb1 expression from a common promoter variant and increased tumor risk after radiation exposure. We propose that a reduced Rb1 expression by common variants in regulatory regions can modify the risk for a malignant transformation of bone cells after radiation exposure.
AU  - Rosemann, M.
AU  - Gonzalez-Vasconcellos, I.
AU  - Domke, T.
AU  - Kuosaite, V.
AU  - Schneider, R.
AU  - Kremer, M.
AU  - Favor, J.
AU  - Nathrath, M.
AU  - Atkinson, M.J.
C1  - 31898
C2  - 34854
CY  - London
TI  - A Rb1 promoter variant with reduced activity contributes to osteosarcoma susceptibility in irradiated mice.
JO  - Mol. Cancer
VL  - 13
IS  - 1
PB  - Biomed Central Ltd
PY  - 2014
ER  - 

TY  - JOUR
AB  - BACKGROUND: CRIP1 (cysteine-rich intestinal protein 1) has been found in several tumor types, its prognostic impact and its role in cellular processes, particularly in breast cancer, are still unclear. METHODS: To elucidate the prognostic impact of CRIP1, we analyzed tissues from 113 primary invasive ductal breast carcinomas using immunohistochemistry. For the functional characterization of CRIP1, its endogenous expression was transiently downregulated in T47D and BT474 breast cancer cells and the effects analyzed by immunoblotting, WST-1 proliferation assay and invasion assay. RESULTS: We found a significant correlation between CRIP1 and HER2 (human epidermal growth factor receptor 2) expression levels (p = 0.016) in tumor tissues. In Kaplan Meier analyses, CRIP1 expression was significantly associated with the distant metastases-free survival of patients, revealing a better prognosis for high CRIP1 expression (p = 0.039). Moreover, in multivariate survival analyses, the expression of CRIP1 was an independent negative prognostic factor, along with the positive prognosticators nodal status and tumor size (p = 0.029). CRIP1 knockdown in the T47D and BT474 breast cancer cell lines led to the increased phosphorylation of MAPK and Akt, to the reduced phosphorylation of cdc2, and to a significantly elevated cell proliferation in vitro (p < 0.001). These results indicate that reduced CRIP1 levels may increase cell proliferation and activate cell growth. In addition, CRIP1 knockdown increased cell invasion in vitro. CONCLUSIONS: Because the lack of CRIP1 expression in breast cancer tissue is significantly associated with a worse prognosis for patients and low endogenous CRIP1 levels in vitro increased the malignant potential of breast cancer cells, we hypothesize that CRIP1 may act as a tumor suppressor in proliferation and invasion processes. Therefore, CRIP1 may be an independent prognostic marker with significant predictive power for use in breast cancer therapy.
AU  - Ludyga, N.
AU  - Englert, S.
AU  - Pflieger, K.
AU  - Rauser, S.
AU  - Braselmann, H.
AU  - Walch, A.K.
AU  - Auer, G.*
AU  - Höfler, H.
AU  - Aubele, M.
C1  - 24260
C2  - 31350
TI  - The impact of Cysteine-Rich Intestinal Protein 1 (CRIP1) in human breast cancer.
JO  - Mol. Cancer
VL  - 12
IS  - 1
PB  - BioMed Central
PY  - 2013
ER  - 

TY  - JOUR
AB  - Aurora-A is a bona-fide oncogene whose expression is associated with genomic instability and malignant transformation. In several types of cancer, gene amplification and/or increased protein levels of Aurora-A are a common feature. RESULTS: In this report, we describe that inhibition of cell proliferation is the main effect observed after transient overexpression of Aurora-A in primary human cells. In addition to the known cell cycle block at the G2/M transition, Aurora-A overexpressing cells fail to overcome the restriction point at the G1/S transition due to diminished RB phosphorylation caused by reduced Cyclin D1 expression. Consequently, overexpression of Cyclin D1 protein is able to override the Aurora-A mediated G1 block. The Aurora-A mediated cell cycle arrest in G2 is not influenced by Cyclin D1 and as a consequence cells accumulate in G2. Upon deactivation of p53 part of the cells evade this premitotic arrest to become aneuploid. CONCLUSION: Our studies describe that an increase of Aurora-A expression levels on its own has a tumor suppressing function, but in combination with the appropriate altered intracellular setting it might exert its oncogenic potential. The presented data indicate that deactivation of the tumor suppressor RB is one of the requirements for overriding a cell cycle checkpoint triggered by increased Aurora-A levels.
AU  - Jantscher, F.*
AU  - Pirker, C.*
AU  - Mayer, C.E.
AU  - Berger, W.*
AU  - Sutterluety, H.*
C1  - 6315
C2  - 28480
TI  - Overexpression of Aurora-A in primary cells interferes with S-phase entry by diminishing Cyclin D1 dependent activities.
JO  - Mol. Cancer
VL  - 10
PB  - Biomed Central Ltd
PY  - 2011
ER  - 

TY  - JOUR
AB  - BACKGROUND: p27Kip1 (p27) is an important negative regulator of the cell cycle and a putative tumor suppressor. The finding that a spontaneous germline frameshift mutation in Cdkn1b (encoding p27) causes the MENX multiple endocrine neoplasia syndrome in the rat provided the first evidence that Cdkn1b is a tumor susceptibility gene for endocrine tumors. Noteworthy, germline p27 mutations were also identified in human patients presenting with endocrine tumors. At present, it is not clear which features of p27 are crucial for this tissue-specific tumor predisposition in both rats and humans. It was shown that the MENX-associated Cdkn1b mutation causes reduced expression of the encoded protein, but the molecular mechanisms are unknown. To better understand the role of p27 in tumor predisposition and to characterize the MENX animal model at the molecular level, a prerequisite for future preclinical studies, we set out to assess the functional properties of the MENX-associated p27 mutant protein (named p27fs177) in vitro and in vivo. RESULTS: In vitro, p27fs177 retains some properties of the wild-type p27 (p27wt) protein: it localizes to the nucleus; it interacts with cyclin-dependent kinases and, to lower extent, with cyclins. In contrast to p27wt, p27fs177 is highly unstable and rapidly degraded in every phase of the cell-cycle, including quiescence. It is in part degraded by Skp2-dependent proteasomal proteolysis, similarly to p27wt. Photobleaching studies showed reduced motility of p27fs177 in the nucleus compared to p27wt, suggesting that in this compartment p27fs177 is part of a multi-protein complex, likely together with the degradation machinery. Studies of primary rat newborn fibroblasts (RNF) established from normal and MENX-affected littermates confirmed the rapid degradation of p27fs177 in vivo which can be rescued by Bortezomib (proteasome inhibitor drug). Overexpression of the negative regulators microRNA-221/222 plays no role in regulating the amount of p27fs177 in RNFs and rat tissues. CONCLUSION: Our findings show that reduced p27 levels, not newly acquired properties, trigger tumor formation in rats, similarly to what has been observed in mice. The molecular characteristics of p27fs177 establish MENX as a useful preclinical model to evaluate compounds that inhibit p27 degradation for their efficacy against endocrine tumors.
AU  - Molatore, S.
AU  - Kiermaier, E.
AU  - Jung, C.B.
AU  - Lee, M.S.
AU  - Pulz, E.
AU  - Höfler, H.
AU  - Atkinson, M.J.
AU  - Pellegata, N.S.
C1  - 4871
C2  - 27455
TI  - Characterization of a naturally-occurring p27 mutation predisposing to multiple endocrine tumors.
JO  - Mol. Cancer
VL  - 9
PB  - BioMed Central Ltd.
PY  - 2010
ER  -