TY  - JOUR
AB  - The response to neoadjuvant therapy can vary widely between individual patients. Histopathological tumor regression grading (TRG) is a strong factor for treatment response and survival prognosis of esophageal adenocarcinoma (EAC) patients following neoadjuvant treatment and surgery. However, TRG systems are usually based on the estimation of residual tumor but do not consider stromal or metabolic changes after treatment. Spatial metabolomics analysis is a powerful tool for molecular tissue phenotyping but has not been used so far in the context of neoadjuvant treatment of esophageal cancer. We used imaging mass spectrometry to assess the potential of spatial metabolomics on tumor and stroma tissue for evaluating therapy response of neoadjuvant-treated EAC patients. With an accuracy of 89.7%, the binary classifier trained on spatial tumor metabolite data proved to be superior for stratifying patients when compared to histopathological response assessment which had an accuracy of 70.5%. Sensitivities and specificities for the poor and favorable survival patient groups ranged from 84.9 to 93.3% using the metabolic classifier and from 62.2 to 78.1% using TRG. The tumor classifier was the only significant prognostic factor (HR 3.38, 95% CI = 1.40-8.12, P = 0.007) when adjusted for clinicopathological parameters such as TRG (HR 1.01, 95% CI = 0.67-1.53, P = 0.968) or stromal classifier (HR 1.856, 95% CI = 0.81-4.25, P = 0.143). The classifier even allowed to further stratify patients within the TRG1-3 categories. The underlying mechanisms of response to treatment has been figured out through network analysis. In summary, metabolic response evaluation outperformed histopathological response evaluation in our study with regard to prognostic stratification. This finding indicates that the metabolic constitution of tumor may have a greater impact on patient survival than the quantity of residual tumor cells or the stroma. This article is protected by copyright. All rights reserved.
AU  - Buck, A.
AU  - Prade, V.M.
AU  - Kunzke, T.
AU  - Feuchtinger, A.
AU  - Kröll, D.*
AU  - Feith, M.*
AU  - Dislich, B.*
AU  - Balluff, B.*
AU  - Langer, R.*
AU  - Walch, A.K.
C1  - 63427
C2  - 51530
CY  - 111 River St, Hoboken 07030-5774, Nj Usa
SP  - 202-213
TI  - Metabolic tumor constitution is superior to tumor regression grading for evaluating response to neoadjuvant therapy of esophageal adenocarcinoma patients.
JO  - J. Pathol.
VL  - 256
IS  - 2
PB  - Wiley
PY  - 2022
SN  - 0022-3417
ER  - 

TY  - JOUR
AB  - Immune checkpoint inhibitors (ICIs) have revolutionized cancer therapy. However, structured knowledge to mitigate a patient's specific risk of developing adverse events are limited. Nevertheless, there is an exponential growth of clinical studies combining conventional therapies such as radiation therapy (RT) with ICIs. Cutaneous reactions are amongst the most common adverse events after monotherapy with either ICIs or RT. So far, little is known about inter-individual differences in the risk of developing severe tissue toxicity after the combination of RT with ICIs, and the underlying biological mechanisms are ill defined. We used experimental models of RT-induced skin injury to analyze skin toxicity after simultaneous application of ICIs. We compared different RT regimens such as fractionated or stereotactic RT with varying dose intensity. Strikingly, we found that simultaneous application of RT and ICIs did not significantly aggravate acute skin injury in two different mouse strains. Detailed examination of long-term tissue damage of the skin revealed similar signs of epidermal hyperplasia, dermal fibrosis, and adnexal atrophy. In summary, we here present the first experimental study demonstrating excellent safety profiles of concurrent treatment with RT and ICIs. These findings will help to interpret the development of adverse events of the skin after radioimmunotherapy and guide the design of new clinical trials and clinical decision making in individual cases. This article is protected by copyright. All rights reserved.
AU  - Lansink Rotgerink, L.*
AU  - Felchle, H.*
AU  - Feuchtinger, A.
AU  - Nefzger, S.M.*
AU  - Walther, C.N.*
AU  - Gissibl, J.*
AU  - Steiger, K.*
AU  - Schmid, T.E.
AU  - Heidegger, S.*
AU  - Combs, S.E.
AU  - Fischer, J.C.*
C1  - 65634
C2  - 52755
SP  - 189-198
TI  - Experimental investigation of skin toxicity after immune checkpoint inhibition in combination with radiation therapy.
JO  - J. Pathol.
VL  - 258
IS  - 2
PY  - 2022
SN  - 0022-3417
ER  - 

TY  - JOUR
AB  - Mitochondria play essential roles in numerous metabolic pathways including the synthesis of adenosine triphosphate through oxidative phosphorylation. Clinically, mitochondrial diseases occur when there is mitochondrial dysfunction - manifesting at any age and affecting any organ system; tissues with high energy requirements, such as muscle and the brain, are often affected. The clinical heterogeneity is parallel to the degree of genetic heterogeneity associated with mitochondrial dysfunction. Around 10% of human genes are predicted to have a mitochondrial function, and defects in over 300 genes are reported to cause mitochondrial disease. Some involve the mitochondrial genome (mtDNA), but the vast majority occur within the nuclear genome. Except for a few specific genetic defects, there remains no cure for mitochondrial diseases which means that a genetic diagnosis is imperative for genetic counselling and the provision of reproductive options for at-risk families. Next-generation sequencing strategies, particularly exome and whole-genome sequencing, have revolutionised mitochondrial diagnostics such that the traditional muscle biopsy has largely been replaced with a minimally-invasive blood sample for an unbiased approach to genetic diagnosis. Where these genomic approaches have not identified a causative defect, or where there is insufficient support for pathogenicity, additional functional investigations are required. The application of supplementary 'omics' technologies, including transcriptomics, proteomics, and metabolomics, has the potential to greatly improve diagnostic strategies. This review aims to demonstrate that, whilst a molecular diagnosis can be achieved for many cases through next-generation sequencing of blood DNA, the use of patient tissues and an integrated, multidisciplinary multi-omics approach is pivotal for diagnosing more challenging cases. Moreover, the analysis of clinically-relevant tissues from affected individuals remains crucial for understanding the molecular mechanisms underlying mitochondrial pathology. This article is protected by copyright. All rights reserved.
AU  - Alston, C.L.*
AU  - Stenton, S.
AU  - Hudson, G.*
AU  - Prokisch, H.
AU  - Taylor, R.W.*
C1  - 61506
C2  - 50323
CY  - 111 River St, Hoboken 07030-5774, Nj Usa
SP  - 430-442
TI  - The genetics of mitochondrial disease: Dissecting mitochondrial pathology using multi-omic pipelines.
JO  - J. Pathol.
VL  - 254
IS  - 4
PB  - Wiley
PY  - 2021
SN  - 0022-3417
ER  - 

TY  - JOUR
AB  - Here, we present an experimental model for human luminal progenitor cells that enables single, primary cells isolated from normal tissue to generate complex branched structures resembling the ductal morphology of low-grade carcinoma of no special type (NST). Thereby, we find that ductal structures are generated through invasive branching morphogenesis via matrix-remodeling and identify reduced actomyosin contractility as a prerequisite for invasion. In addition, we show that knockout of E-cadherin causes a dissolution of duct formation as observed in invasive lobular carcinoma (ILC), a subtype of invasive carcinomas where E-cadherin function is frequently lost. Thus, our model shows that invasive capacity can be elicited from normal luminal cells in specific environments which results in low-grade NST morphology. This assay offers a platform to investigate the dynamics of luminal cell invasion and unravel the impact of genetic and non-genetic aberrations on invasive morphology.
AU  - Ganz, H.M.
AU  - Buchmann, B.*
AU  - Engelbrecht, L.K.*
AU  - Jesinghaus, M.*
AU  - Eichelberger, L.*
AU  - Gabka, C.J.*
AU  - Schmidt, G.P.*
AU  - Muckenhuber, A.*
AU  - Weichert, W.*
AU  - Bausch, A.R.*
AU  - Scheel, C.
C1  - 62968
C2  - 51123
CY  - 111 River St, Hoboken 07030-5774, Nj Usa
TI  - Generation of ductal organoids from normal mammary luminal cells reveals invasive potential.
JO  - J. Pathol.
PB  - Wiley
PY  - 2021
SN  - 0022-3417
ER  - 

TY  - JOUR
AB  - The Epstein-Barr virus (EBV) is found almost exclusively in the activated B-cell (ABC) subtype of diffuse large B-cell lymphoma (DLBCL), yet its contribution to this tumour remains poorly understood. We have focused on the EBV-encoded latent membrane protein-1 (LMP1), a constitutively activated CD40 homologue expressed in almost all EBV-positive DLBCLs and which can disrupt germinal centre (GC) formation and drive lymphomagenesis in mice. Comparison of the transcriptional changes that follow LMP1 expression with those that follow transient CD40 signalling in human GC B cells enabled us to define pathogenic targets of LMP1 aberrantly expressed in ABC-DLBCL. These included the down-regulation of S1PR2, a sphingosine-1-phosphate (S1P) receptor that is transcriptionally down-regulated in ABC-DLBCL, and when genetically ablated leads to DLBCL in mice. Consistent with this, we found that LMP1-expressing primary ABC-DLBCLs were significantly more likely to lack S1PR2 expression than were LMP1-negative tumours. Furthermore, we showed that the down-regulation of S1PR2 by LMP1 drives a signalling loop leading to constitutive activation of the phosphatidylinositol-3-kinase (PI3-K) pathway. Finally, core LMP1-PI3-K targets were enriched for lymphoma-related transcription factors and genes associated with shorter overall survival in patients with ABC-DLBCL. Our data identify a novel function for LMP1 in aggressive DLBCL. Copyright (c) 2019 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
AU  - Vockerodt, M.*
AU  - Vrzalikova, K.*
AU  - Ibrahim, M.*
AU  - Nagy, E.*
AU  - Margielewska, S.*
AU  - Hollows, R.*
AU  - Lupino, L.*
AU  - Tooze, R.*
AU  - Care, M.*
AU  - Simmons, W.*
AU  - Schrader, A.*
AU  - Perry, T.*
AU  - Abdullah, M.*
AU  - Foster, S.*
AU  - Reynolds, G.*
AU  - Dowell, A.*
AU  - Rudski, Z.*
AU  - Krappmann, D.
AU  - Kube, D.*
AU  - Woodman, C.*
AU  - Wei, W.*
AU  - Taylor, G.*
AU  - Murray, P.G.*
C1  - 55311
C2  - 46305
CY  - 111 River St, Hoboken 07030-5774, Nj Usa
SP  - 142-154
TI  - Regulation of S1PR2 by the EBV oncogene LMP1 in aggressive ABC-subtype diffuse large B-cell lymphoma.
JO  - J. Pathol.
VL  - 248
IS  - 2
PB  - Wiley
PY  - 2019
SN  - 0022-3417
ER  - 

TY  - JOUR
AB  - Oesophageal squamous cell carcinomas and oesophageal adenocarcinomas display distinct patterns of ErbB expression and dimers. The functional effects of specific ErbB homo- or heterodimers on oesophageal (cancer) cell behaviour, particularly invasion of early carcinogenesis remains unknown. Here, a new cellular model system for controlled activation of EGFR or HER2 and EGFR/HER2 or HER2/HER3 homo- and heterodimers was studied in non-neoplastic squamous oesophageal epithelial Het-1A cells. EGFR, HER2 and HER3 intracellular domains (ICDs) were fused to dimerization domains (DmrA / DmrA and DmrC), and transduced into Het-1A cells lacking ErbB expression. Dimerization of EGFR, HER2 or EGFR/HER2, HER2/HER3 ICDs was induced by synthetic ligands (A/A or A/C dimerizers). This was accompanied by phosphorylation of the respective EGFR, HER2 and HER3 ICDs and activation of distinct down-stream signalling pathways, such as PLCγ1, Akt, STAT and Src family kinases. Phenotypically, ErbB homo- and heterodimers caused cell rounding and non-apoptotic blebbing in EGFR/HER2 and HER2/HER3 heterodimer cells. In a Transwell assay, cell migration velocity was elevated in HER2-dimer as compared to empty vector cells. In addition, HER2-dimer cells showed in increased cell invasion, reaching significance for induced HER2/HER3 heterodimers (p=0.015). Importantly, in three-dimensional organotypic cultures, empty vector cells grew as a superficial cell layer, resembling oesophageal squamous epithelium. In contrast, induced HER2-dimer cells (HER2 homodimers) were highly invasive into the matrix and formed cell clusters. This was associated with partial loss of CK7 (when HER2 homodimers were modelled) and p63 (when EGFR/HER2 heterodimers were modelled), which suggests a change or loss of squamous cell differentiation. Controlled activation of specific EGFR, HER2 and HER3 homo- and heterodimers caused oesophageal squamous epithelial cell migration and/or invasion, especially in a three dimensional microenvironment, thereby functionally identifying ErbB homo- and heterodimers as important drivers of oesophageal carcinogenesis.
AU  - Fichter, C.D.*
AU  - Przypadlo, C.M.*
AU  - Buck, A.
AU  - Herbener, N.*
AU  - Riedel, B.*
AU  - Schäfer, L.*
AU  - Nakagawa, H.*
AU  - Walch, A.K.
AU  - Reinheckel, T.*
AU  - Werner, M.*
AU  - Lassmann, S.*
C1  - 51963
C2  - 43624
CY  - Hoboken
SP  - 481-495
TI  - A new model system identifies EGFR/HER2 and HER2/HER3 heterodimers as potent inducers of oesophageal epithelial cell invasion.
JO  - J. Pathol.
VL  - 243
IS  - 4
PB  - Wiley
PY  - 2017
SN  - 0022-3417
ER  - 

TY  - JOUR
AB  - We present the first analytical approach to demonstrate the in situ imaging of metabolites from formalin-fixed paraffin-embedded (FFPE) human tissue samples. Using high-resolution Matrix-Assisted Laser Desorption/Ionization Fourier-Transform Ion Cyclotron Resonance Mass Spectrometry Imaging (MALDI-FT-ICR MSI), we conducted a proof of principle experiment comparing metabolite measurements from FFPE and fresh frozen tissue sections, and found an overlap of 72% amongst 1700 m/z species. In particular, we observed conservation of biomedically relevant information at the metabolite level in FFPE tissues. In biomedical applications, we analysed tissues from 350 different cancer patients, and were able to discriminate between normal and tumour tissues, and different tumours from the same organ, and found an independent prognostic factor for patient survival. This study demonstrates the ability to measure metabolites in FFPE tissues using MALDI-FT-ICR MSI, which can then be assigned to histology and clinical parameters. Our approach is a major technical, histochemical and clinicopathological advance that highlights the potential for investigating diseases in archived FFPE tissues.
AU  - Buck, A.
AU  - Ly, A.
AU  - Balluff, B.*
AU  - Sun, N.
AU  - Gorzolka, K.
AU  - Feuchtinger, A.
AU  - Janssen, K.P.*
AU  - Kuppen, P.J.*
AU  - van de Velde, C.J.*
AU  - Weirich, G.*
AU  - Erlmeier, F.*
AU  - Langer, R.*
AU  - Aubele, M.
AU  - Zitzelsberger, H.
AU  - Aichler, M.
AU  - Walch, A.K.
C1  - 44840
C2  - 37034
CY  - Hoboken
SP  - 123–132
TI  - High-resolution MALDI-FT-ICR MS imaging for the analysis of metabolites from formalin-fixed paraffin-embedded clinical tissue samples.
JO  - J. Pathol.
VL  - 237
IS  - 1
PB  - Wiley-blackwell
PY  - 2015
SN  - 0022-3417
ER  - 

TY  - JOUR
AB  - Chronic Hepatitis B virus (HBV) infection remains the number one risk factor for hepatocellular carcinoma (HCC), accounting for more than 600000 deaths per year. Despite highly effective antiviral treatment options, chronic hepatitis B (CHB), subsequent end stage liver disease and HCC development remain a major challenge worldwide. In CHB, liver damage is mainly caused by the influx of immune cells and destruction of infected hepatocytes causing necroinflammation. Treatment with nucleos(t)id analogs can effectively suppress HBV replication in patients with CHB and thus decrease the risk for HCC development. Nevertheless, the risk of HCC in treated patients showing sufficient suppression of HBV DNA replication is significantly higher than in patients with inactive CHB, regardless of the presence of baseline liver cirrhosis, suggesting direct, long lasting, predisposing effects of HBV. Direct oncogenic effects of HBV include integration in the host genome leading to deletions, cis- / trans-activation, translocations, the production of fusion transcripts, and generalized genomic instability, as well as pleiotropic effects of viral transcripts (HBsAg and HBx). Analysis of these viral factors in active surveillance may allow early identification of high-risk patients, and their integration into a molecular classification of HCC subtypes might help in the development of novel therapeutic approaches.
AU  - Ringelhan, M.
AU  - Protzer, U.
AU  - O'Connor, T.*
AU  - Heikenwälder, M.
C1  - 32120
C2  - 34970
SP  - 355-367
TI  - The direct and indirect role of HBV in liver cancer: Prospective markers for HCC-screening and potential therapeutic targets.
JO  - J. Pathol.
VL  - 235
IS  - 2
PY  - 2015
SN  - 0022-3417
ER  - 

TY  - JOUR
AB  - Barrett's oesophagus is a metaplastic change, such that the normal squamous epithelial lining of the oesophagus is replaced by specialised columnar lined epithelium. Barrett's oesophagus is clinically significant and has a high health economic impact as it is associated with heightened risk of progression to oesophageal adenocarcinoma. This review discusses the pathogenesis of Barrett's oesophagus with an emphasis on the underlying molecular events.
AU  - Aichler, M.
AU  - Walch, A.K.
C1  - 28470
C2  - 33410
SP  - 383-385
TI  - In brief: The (molecular) pathogenesis of Barrett's oesophagus.
JO  - J. Pathol.
VL  - 232
IS  - 4
PB  - Wiley-Blackwell
PY  - 2014
SN  - 0022-3417
ER  - 

TY  - JOUR
AB  - An essential and so far unresolved factor influencing the evolution of cancer and the clinical management of patients is intra-tumor clonal and phenotypic heterogeneity. However, the de novo identification of tumor subpopulations is a so far challenging, if not an unresolved, task. Here we present the first systematic approach for the de novo discovery of clinically detrimental molecular tumor subpopulations. In this proof-of-principle study, spatially-resolved, tumor-specific mass spectra were acquired using matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry from tissues of 63 gastric carcinoma and 32 breast carcinoma patients. The mass spectra, representing the proteomic heterogeneity within tumor areas, were grouped by a corroborated statistical clustering algorithm in order to obtain segmentation maps of molecularly distinct regions. These regions were presumed to represent different phenotypic tumor subpopulations. This was confirmed by linking the presence of these tumor subpopulations to the patients' clinical data. This revealed several of the detected tumor subpopulations to be associated with a different overall survival of the gastric cancer patients (P = 0.025) and the presence of locoregional metastases in patients with breast cancer (P = 0.036). The procedure presented is generic and opens novel options in cancer research as it reveals microscopically indistinct tumor subpopulations that have an adverse impact on clinical outcome. This enables their further molecular characterization for deeper insights into the biological processes of cancer which may finally lead to new targeted therapies.
AU  - Balluff, B.*
AU  - Frese, C.K.*
AU  - Maier, S.K.
AU  - Schöne, C.
AU  - Kuster, B.*
AU  - Schmitt, M.*
AU  - Aubele, M.
AU  - Höfler, H.
AU  - Deelder, A.M.*
AU  - Heck, A.J.*
AU  - Hogendoorn, P.C.W.*
AU  - Morreau, J.*
AU  - Altelaar, A.F.*
AU  - Walch, A.K.
AU  - McDonnell, L.A.*
C1  - 32247
C2  - 34987
SP  - 3-13
TI  - De novo discovery of phenotypic intra-tumor heterogeneity using imaging mass spectrometry.
JO  - J. Pathol.
VL  - 235
IS  - 1
PY  - 2014
SN  - 0022-3417
ER  - 

TY  - JOUR
AB  - Chemotherapeutic drugs kill cancer cells, but it is unclear why this happens in responding patients but not in non-responders. Proteomic profiles of patients with oesophageal adenocarcinoma may be helpful in predicting response and selecting more effective treatment strategies. In this study, pre-therapeutic oesophageal adenocarcinoma biopsies were analysed for proteomic changes associated with response to chemotherapy by MALDI imaging mass spectrometry. Resulting candidate proteins were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and investigated for functional relevance in vitro. Clinical impact was validated in pretherapeutic biopsies from an independent patient cohort. Studies on the incidence of these defects in other solid tumours were included. We discovered that clinical response to cisplatin correlated with pre-existing defects in the mitochondrial respiratory chain complexes of cancer cells, caused by loss of specific cytochrome c oxidase (COX) subunits. Knockdown of a COX protein altered chemosensitivity in vitro, increasing the propensity of cancer cells to undergo cell death following cisplatin treatment. In an independent validation, patients with reduced COX protein expression prior to treatment exhibited favourable clinical outcomes to chemotherapy, whereas tumours with unchanged COX expression were chemoresistant. In conclusion, previously undiscovered pre-existing defects in mitochondrial respiratory complexes cause cancer cells to become chemosensitive: mitochondrial defects lower the cells' threshold for undergoing cell death in response to cisplatin. By contrast, cancer cells with intact mitochondrial respiratory complexes are chemoresistant and have a high threshold for cisplatin-induced cell death. This connection between mitochondrial respiration and chemosensitivity is relevant to anticancer therapeutics that target the mitochondrial electron transport chain.
AU  - Aichler, M.
AU  - Elsner, M.
AU  - Ludyga, N.
AU  - Feuchtinger, A.
AU  - Zangen, V.
AU  - Maier, S.K.
AU  - Balluff, B.
AU  - Schöne, C.
AU  - Hierber, L.
AU  - Braselmann, H.
AU  - Meding, S.
AU  - Rauser, S.
AU  - Zischka, H.
AU  - Aubele, M.
AU  - Schmitt, M.*
AU  - Feith, M.*
AU  - Hauck, S.M.
AU  - Ueffing, M.
AU  - Langer, R.*
AU  - Kuester, B.*
AU  - Zitzelsberger, H.
AU  - Höfler, H.
AU  - Walch, A.K.
C1  - 24004
C2  - 31307
SP  - 410-419
TI  - Clinical response to chemotherapy in oesophageal adenocarcinoma patients is linked to defects in mitochondria.
JO  - J. Pathol.
VL  - 230
IS  - 4
PB  - Wiley-Blackwell
PY  - 2013
SN  - 0022-3417
ER  - 

TY  - JOUR
AB  - Metastatic spread in Ewing sarcomas (ES) is frequent and haematogenous. G-protein coupled receptor 64 (GPR64), an orphan receptor with normal expression restricted to human epididymis is specifically over-expressed in ES among sarcoma, but also up-regulated in a number of carcinomas derived from prostate, kidney or lung. Inhibition of GPR64 expression in ES by RNA interference impaired colony formation in vitro and suppressed local tumour growth and metastasis in Rag2/C/ mice. Microarray analysis after GPR64 knock down revealed a GPR64-mediated repression of genes involved in neuronal development like SLIT, drosophila, homolog of, 2 (SLIT2), and genes regulating transcription including pre-B cell leukemia homeobox2 (PBX2). Concurrently, the suppression of GPR64 increased ES susceptibility to TRAIL induced apoptosis. Moreover, a GPR64-mediated induction of placental growth factor (PGF) in ES was observed. PGF suppression by RNA interference resulted in a reduction of metastatic growth similar to that observed after GPR64 knock down. Importantly, inhibition of GPR64 as well as PGF expression was associated with a reduced expression of matrix metalloproteinase (MMP) 1 and invasiveness in vitro. Furthermore, MMP1 knock down abrogated lung metastasis in Rag2/C/ mice. Thus, GPR64 expression in ES maintains an immature phenotype that is less sensitive to TRAIL-induced apoptosis and via its up-regulation of PGF and MMP1 orchestrates and promotes invasiveness and metastatic spread.
AU  - Richter, G.H.S.*
AU  - Fasan, A.*
AU  - Hauer, K.*
AU  - Grunewald, T.G.P.*
AU  - Berns, C.*
AU  - Rössler, S.*
AU  - Naumann, I.*
AU  - Staege, M.S.*
AU  - Fulda, S.*
AU  - Esposito, I.
AU  - Burdach, S.*
C1  - 24440
C2  - 31558
SP  - 70-81
TI  - G-protein coupled receptor 64 promotes invasiveness and metastasis in Ewing sarcomas through PGF and MMP1.
JO  - J. Pathol.
VL  - 230
IS  - 1
PB  - Wiley-Blackwell
PY  - 2013
SN  - 0022-3417
ER  - 

TY  - JOUR
AB  - Pancreatic ductal adenocarcinoma (PDAC) and its precursor lesions, pancreatic intraepithelial neoplasia, (PanIN), display a ductal phenotype. However, there is evidence in genetically defined mouse models for PDAC harbouring a mutated kras under the control of a pancreas-specific promoter that ductal cancer might arise in the centroacinar-acinar region, possibly through a process of acinar-ductal metaplasia (ADM). In order to further elucidate this model of PDAC development, an extensive expression analysis and molecular characterization of the putative and already established (PanIN) precursor lesions was performed in the Kras(G12D/+) ;Ptf1a-Cre(ex1/+) mouse model and in human tissues, focusing on lineage markers, developmental pathways, cell cycle regulators, apomucins and stromal activation markers. The results of this study show that areas of ADM are very frequent in the murine and human pancreas and represent regions of increased proliferation of cells with precursor potential. Moreover, atypical flat lesions originating in areas of ADM are the most probable precursors of PDAC in the Kras(G12D/+) ;Ptf1a-cre(ex1/+) mice and similar lesions were also found in the pancreas of three patients with a strong family history of PDAC. In conclusion, PDAC development in Kras(G12D/+) ;Ptf1a-Cre(ex1/+) mice starts from ADM and a similar process might also take place in patients with a strong family history of PDAC.
AU  - Aichler, M.
AU  - Seiler, C.*
AU  - Tost, M.
AU  - Siveke, J.*
AU  - Mazur, P.K.*
AU  - Da Silva-Buttkus, P.
AU  - Bartsch, D.K.*
AU  - Langer, P.*
AU  - Chiblak, S.*
AU  - Dürr, A.*
AU  - Höfler, H.*
AU  - Klöppel, G.*
AU  - Müller-Decker, K.*
AU  - Brielmeier, M.
AU  - Esposito, I.
C1  - 6755
C2  - 29211
SP  - 723-734
TI  - Origin of pancreatic ductal adenocarcinoma from atypical flat lesions: A comparative study in transgenic mice and human tissues.
JO  - J. Pathol.
VL  - 226
IS  - 5
PB  - Wiley-Blackwell
PY  - 2012
SN  - 0022-3417
ER  - 

TY  - JOUR
AB  - Regional lymph node metastasis negatively affects prognosis in colon cancer patients. The molecular processes leading to regional lymph node metastasis are only partially understood and proteomic markers for metastasis are still scarce. Therefore, a tissue-based proteomic approach was undertaken for identifying proteins associated with regional lymph node metastasis. Two complementary tissue-based proteomic methods have been employed. MALDI imaging was used for identifying small proteins (≤25 kDa) in situ and label-free quantitative proteomics was used for identifying larger proteins. A tissue cohort comprising primary colon tumours without metastasis (UICC II, pN0, n = 21) and with lymph node metastasis (UICC III, pN2, n = 33) was analysed. Subsequent validation of identified proteins was done by immunohistochemical staining on an independent tissue cohort consisting of primary colon tumour specimens (n = 168). MALDI imaging yielded ten discriminating m/z species, and label-free quantitative proteomics 28 proteins. Two MALDI imaging-derived candidate proteins (FXYD3 and S100A11) and one from the label-free quantitative proteomics (GSTM3) were validated on the independent tissue cohort. All three markers correlated significantly with regional lymph node metastasis: FXYD3 (p = 0.0110), S100A11 (p = 0.0071), and GSTM3 (p = 0.0173). FXYD3 and S100A11 were more highly expressed in UICC II patient tumour tissues. GSTM3 was more highly expressed in UICC III patient tumour tissues. By our tissue-based proteomic approach, we could identify a large panel of proteins which are associated with regional lymph node metastasis and which have not been described so far. Here we show that novel markers for regional lymp.
AU  - Meding, S.
AU  - Balluff, B.
AU  - Elsner, M.
AU  - Schöne, C.
AU  - Rauser, S.
AU  - Nitsche, U.*
AU  - Maak, M.*
AU  - Schäfer, A.
AU  - Hauck, S.M.
AU  - Ueffing, M.
AU  - Langer, R.*
AU  - Höfler, H.
AU  - Friess, H.*
AU  - Rosenberg, R.*
AU  - Walch, A.K.
C1  - 10958
C2  - 30452
SP  - 459-470
TI  - Tissue-based proteomics reveals FXYD3, S100A11 and GSTM3 as novel markers for regional lymph node metastasis in colon cancer.
JO  - J. Pathol.
VL  - 228
IS  - 4
PB  - Wiley-Blackwell
PY  - 2012
SN  - 0022-3417
ER  - 

TY  - JOUR
AB  - The urokinase-type plasminogen activator (uPA) and the main uPA inhibitor PAI-1 play important roles in cell migration and invasion in both physiological and pathological contexts. Both factors are clinically applicable predictive markers in node-negative breast cancer patients that are used to stratify patients for adjuvant chemotherapy. In addition to their classical functions in plasmin regulation, both factors are key components in cancer-related cell signalling. Such signalling cascades are well described in cell culture systems, but a better understanding of uPA- and PAI-1-associated signalling networks in clinical tissues is needed. We examined the expression of uPA, PAI-1, and 21 signalling molecules in 201 primary breast cancer tissues using protein microarrays. Expression of uPA was significantly correlated with the expression of ERK and Stat3, while expression of PAI-1 was correlated with the uPA receptor and Akt activation, presumably via integrin and HER-receptor signalling. Analysis of uPA expression did not reveal any significant correlation with staging, grading or age of the patients. The PAI-1 expression was correlated with nodal stage. Network monitoring for uPA and PAI-1 in breast cancer reveals interactions with main signalling cascades and extends the findings from cell culture experiments. Our results reveal possible mechanisms underlying cancer development.
AU  - Wolff, C.*
AU  - Malinowsky, K.*
AU  - Berg, D.*
AU  - Schragner, K.*
AU  - Schuster, T:*
AU  - Walch, A.K.
AU  - Bronger, H.*
AU  - Höfler, H.
AU  - Becker, K.F.*
C1  - 2162
C2  - 27712
SP  - 54-63
TI  - Signalling networks associated with urokinase-type plasminogen activator (uPA) and its inhibitor PAI-1 in breast cancer tissues: New insights from protein microarray analysis.
JO  - J. Pathol.
VL  - 223
IS  - 1
PB  - Wiley-Blackwell
PY  - 2011
SN  - 0022-3417
ER  - 

TY  - JOUR
AB  - The generation of new mouse models of human disease is accelerating rapidly, due to the completion of whole-genome sequencing efforts and technological advances in the manipulation of the mouse genome. We sought to investigate manpower issues in the provision of histopathology expertise for mouse functional genomics and compared this to the perceived demand from principal investigators (PIs). Through the European Commission (EC)-funded PRIME pathology training initiative, two questionnaires were devised to collect information from pathologists and EC-funded PIs on the current provision of mouse histopathology expertise in Europe and the demands for this service. We find that pathological analysis is being performed almost exclusively by professionally qualified pathologists, generally employed in clinical diagnostic posts, where the work is undertaken as collaboration outside of their contractual commitments but without previous training in veterinary or comparative pathology. The results indicate that there is a lack of both trainees and provision of specialist training in this field. Unsurprisingly, the availability of diagnostic expertise and advice falls far short of the number of genetically engineered mice (GEM) being generated for analysis. We analyse these results with reference to previous studies and discuss solutions for the future recruitment, training and funding for pathologists in mouse functional genomics in Europe.
AU  - Warren, M.V.*
AU  - Studley, M.L.*
AU  - Dubus, P.*
AU  - Fiette, L.*
AU  - Rozell, B.*
AU  - Quintanilla-Martinez, L.
AU  - Raspa, M.*
AU  - Breuer, M.*
AU  - Song, J.Y.*
AU  - Gates, H.*
AU  - Brown, S.D.*
AU  - Schofield, P.N.*
C1  - 872
C2  - 25899
SP  - 4-13
TI  - An impending crisis in the provision of histopathology expertise for mouse functional genomics.
JO  - J. Pathol.
VL  - 217
IS  - 1
PB  - Wiley-Blackwell
PY  - 2009
SN  - 0022-3417
ER  - 

TY  - JOUR
AB  - Owing to its cross-linking effects, it is currently believed that formalin fixation of routinely processed tissues in the clinic prevents protein extraction and profiling. The aim of our study was to develop a robust, fast, standardized, and easy to use technique for the solubilization of non-degraded, full length, and immunoreactive proteins from formalin-fixed tissues for western blot and protein microarray analysis. Sections of routinely processed formalin-fixed and paraffin-embedded tissues of various origin were analysed. After deparaffination, tissues were manually dissected from the slides and transferred into an optimized protein extraction buffer system. Proteins were solubilized and subsequently analysed by western blot and reverse phase protein microarrays. We succeeded in isolating non-degraded, soluble, and immunoreactive proteins from routinely processed formalin-fixed tissues. We were able to detect membrane, cytoplasmic and nuclear proteins at the expected molecular weight. No differences were found in the protein yield and protein abundances between fresh frozen and formalin-fixed tissues. Using western blots and reverse phase protein microarrays, the receptor tyrosine kinase HER2, an important protein target for antibody based cancer treatment, was reliably measured in formalin-fixed breast cancer biopsy samples when compared with measurement by immunohistochemistry and fluorescence in situ hybridization; remarkably, immunohistochemically equivocal cases (score 2 + ) can be categorized according to HER2 protein abundance. Our new clinically orientated multiplexed protein measurement system may be generally applicable to determine the relative abundances of known disease-related proteins in small amounts of routinely processed formalin-fixed tissue samples for research and diagnosis. This technique may also be used to identify, characterize, and validate known and new protein markers in a variety of human diseases.
AU  - Becker, K.-F.*
AU  - Schott, C.*
AU  - Hipp, S.*
AU  - Metzger, V.*
AU  - Porschewski, P.*
AU  - Beck, R.*
AU  - Nährig, J.*
AU  - Becker, I.*
AU  - Höfler, H.
C1  - 2524
C2  - 24688
SP  - 370-378
TI  - Quantitative protein analysis from formalin-fixed tissues: Implications for translational clinical research and nanoscale molecular diagnosis.
JO  - J. Pathol.
VL  - 211
IS  - 3
PB  - Wiley-Blackwell
PY  - 2007
SN  - 0022-3417
ER  - 

TY  - JOUR
AU  - Gerlach, U.*
AU  - Kayser, G.*
AU  - Walch, A.K.
AU  - Hopt, U.*
AU  - Schulte-Mönting, J.*
AU  - Werner, M.*
AU  - Lassmann, S.*
C1  - 1408
C2  - 23545
SP  - 462-472
TI  - Centrosome-, chromosomal-passenger- and cell-cycle-associated mRNAs are differentially regulated in the development of sporadic colorectal cancer.
JO  - J. Pathol.
VL  - 208
PY  - 2006
SN  - 0022-3417
ER  - 

TY  - JOUR
AU  - Hennard, C.
AU  - Pfuhl, T.*
AU  - Buettner, M.*
AU  - Becker, K.-H.*
AU  - Knöfel, T.
AU  - Middeldorp, J.*
AU  - Kremmer, E.
AU  - Niedobitek, G.*
AU  - Grässer, F.A.*
C1  - 389
C2  - 23891
SP  - 430-435
TI  - The antibody 2B4 directed against the Epstein-Barr virus (EBV)-encoded nuclear antigen I (EBNA I) detects MAGE-4: Implications for studies on the EBV association of human cancers.
JO  - J. Pathol.
VL  - 209
PB  - Wiley
PY  - 2006
SN  - 0022-3417
ER  - 

TY  - JOUR
AU  - Schreck, S.*
AU  - Buettner, M.*
AU  - Kremmer, E.
AU  - Bogdan, M.*
AU  - Herbst, H.*
AU  - Niedobitek, G.*
C1  - 2632
C2  - 23892
SP  - 26-31
TI  - Activation-induced cytidine deaminase (AID) is expressed in normal spermatogenesis but only infrequently in testicular germ cell tumours.
JO  - J. Pathol.
VL  - 210
PB  - Wiley
PY  - 2006
SN  - 0022-3417
ER  - 

TY  - JOUR
AU  - Born, M.
AU  - Quintanilla-Fend, L.
AU  - Braselmann, H.
AU  - Reich, U.
AU  - Richter, M.
AU  - Hutzler, P.
AU  - Aubele, M.
C1  - 3060
C2  - 22597
SP  - 592-596
TI  - Simultaneous over-expression of the Her2/neu and PTK6 tyrosine kinases in archival invasive ductal breast carcinomas.
JO  - J. Pathol.
VL  - 205
PB  - Wiley
PY  - 2005
SN  - 0022-3417
ER  - 

TY  - JOUR
AU  - Greiner, A.*
AU  - Tobollik, S.*
AU  - Buettner, M.*
AU  - Jungnickel, B.
AU  - Herrmann, K.*
AU  - Kremmer, E.
AU  - Niedobitek, G.*
C1  - 1929
C2  - 22599
SP  - 541-547
TI  - Differential expression of activation-induced cytidine deaminase (AID) in nodular lymphocyte-predominant and classical Hodgkin lymphoma.
JO  - J. Pathol.
VL  - 205
PB  - Wiley
PY  - 2005
SN  - 0022-3417
ER  - 

TY  - JOUR
AU  - Kremer, M.*
AU  - Ott, G.*
AU  - Nathrath, M.*
AU  - Specht, K.*
AU  - Stecker, K.*
AU  - Alexiou, Ch.*
AU  - Quintanilla-Martinez, L.*
AU  - Fend, F.*
C1  - 4310
C2  - 23198
SP  - 92-101
TI  - Primary extramedullary plasmacytoma and multiple myeloma: Phenotypic differences revealed by immunohistochemical analysis.
JO  - J. Pathol.
VL  - 205
PB  - Wiley
PY  - 2005
SN  - 0022-3417
ER  - 

TY  - JOUR
AU  - Albrecht, B.*
AU  - Hausmann, M.*
AU  - Zitzelsberger, H.
AU  - Stein, H.*
AU  - Siewert, J.R.*
AU  - Hopt, U.*
AU  - Langer, R.*
AU  - Höfler, H.
AU  - Werner, M.*
AU  - Walch, A.K.
C1  - 4617
C2  - 22142
SP  - 780-788
TI  - Array-based comparative genomic hybridization for the detection of DNA sequence copy number changes in Barrett's adenocarcinoma.
JO  - J. Pathol.
VL  - 203
PB  - Wiley
PY  - 2004
SN  - 0022-3417
ER  - 

TY  - JOUR
AU  - Heussinger, N.*
AU  - Büttner, M.*
AU  - Ott, G.*
AU  - Brachtel, E.*
AU  - Pilch, B.Z.*
AU  - Kremmer, E.
AU  - Niedobitek, G.*
C1  - 3413
C2  - 22099
SP  - 696-699
TI  - Expression of the Epstein-Barr virus (EBV)-encoded latent membrane protein 2A (LMP2A) in EBV-associated nasopharyngeal carcinoma.
JO  - J. Pathol.
VL  - 203
PB  - Wiley
PY  - 2004
SN  - 0022-3417
ER  - 

TY  - JOUR
AU  - Kammerer, R.*
AU  - Riesenberg, R.*
AU  - Weiler, C.*
AU  - Lohmann, J.*
AU  - Schleypen, J.S.
AU  - Zimmermann, W.*
C1  - 5289
C2  - 22382
SP  - 258-267
TI  - The tumour suppressor gene CEACAMI is completely but reversibly downregulated in renal cell carcinoma.
JO  - J. Pathol.
VL  - 204
PB  - Wiley
PY  - 2004
SN  - 0022-3417
ER  - 

TY  - JOUR
AU  - Kappler, R.*
AU  - Calzada-Wack, J.
AU  - Schnitzbauer, U.
AU  - Koleva, M.*
AU  - Herwig, A.*
AU  - Piontek, G.*
AU  - Graedler, F.
AU  - Adamski, J.
AU  - Heinzmann, U.
AU  - Schlegel, J.*
AU  - Hemmerlein, B.*
AU  - Quintanilla-Martinez, L.
AU  - Hahn, H.*
C1  - 10048
C2  - 21072
SP  - 348-356
TI  - Molecular characterization of Patched-associated rhabdomyosarcoma.
JO  - J. Pathol.
VL  - 200
PB  - Wiley
PY  - 2003
SN  - 0022-3417
ER  - 

TY  - JOUR
AB  - Immunohistochemical analysis has been used to show that expression of the homophilic cell-to-cell adhesion molecule, E-cadherin, is frequently altered in human cancers, including gastric and breast carcinoma. Besides genetic down-regulation, structural mutations such as in-frame deletions of exon 8 and exon 9 were frequently found; these may affect the binding of monoclonal antibodies used for immunohistochemical analysis. In this study it was found that antibodies HECD-1 and E9, two monoclonal antibodies often used in E-cadherin immunoanalysis, react with epitopes present at least in part in exon 8 and exon 9, respectively. This study generated and characterized a mutation-specific monoclonal antibody, E-cad delta 8-1, reacting with the mutant protein lacking exon 8 but not with the wild-type molecule. By using E-cad delta 8-1 and HECD-1, it was possible separately to analyse the immunoreactivity of mutant and normal E-cadherin proteins, respectively, in an allele-specific manner in archival material. A similar analysis was performed using E9 and the previously characterized mutation-specific antibody E-cad delta 9-1. Typically, in gastric and breast cancer harbouring E-cadherin splice site gene mutations, the mutant proteins were expressed but the wild-type protein was not detected in malignant tissues. These results indicate that variant-specific monoclonal antibodies can be used to identify differentially expressed E-cadherin proteins. For immunohistochemical analysis of E-cadherin, at least two different monoclonal antibodies should be used to exclude alterations of the epitopes resulting in failure to detect a mutant protein.
AU  - Becker, K.-F.
AU  - Kremmer, E.
AU  - Eulitz, M.
AU  - Schulz, St.*
AU  - Mages, J.
AU  - Handschuh, G.*
AU  - Wheelock, M.J.*
AU  - Cleton-Jansen, A.-M.*
AU  - Höfler, H.
AU  - Becker, I.*
C1  - 10047
C2  - 20338
SP  - 567-574
TI  - Functional allelic loss detected at the protein level in archival human tumours using allele-specific E-cadherin monoclonal antibodies.
JO  - J. Pathol.
VL  - 197
PB  - Wiley
PY  - 2002
SN  - 0022-3417
ER  - 

TY  - JOUR
AU  - Gerber, J.-K.
AU  - Richter, T.*
AU  - Kremmer, E.
AU  - Adamski, J.
AU  - Höfler, H.
AU  - Balling, R.
AU  - Peters, H.*
C1  - 22002
C2  - 20544
SP  - 293-297
TI  - Progressive loss of PAX9 expression correlates with increasing malignancy of dysplastic and cancerous epithelium of the human oesophagus.
JO  - J. Pathol.
VL  - 197
PB  - Wiley
PY  - 2002
SN  - 0022-3417
ER  - 

TY  - JOUR
AU  - Huss, R.*
AU  - Weissinger, E.M.
AU  - Lange, C.
AU  - Gatsios, P.*
AU  - Eissner, G.*
AU  - Kolb, H.-J.
AU  - Diebold, J.*
AU  - Heinrich, P.C.*
AU  - Graeve, L.*
C1  - 21672
C2  - 19852
SP  - 363-372
TI  - In vitro-generated stem cell leukaemia showing altered cell cycle progression with distinct signalling of the tyrosine-phosphorylated rasGAP-associated p62dok protein.
JO  - J. Pathol.
VL  - 192
PB  - Wiley
PY  - 2000
SN  - 0022-3417
ER  - 

TY  - JOUR
AB  - Fluorescence in situ hybridization (FISH) using chromosome-specific a- satellite DNA probes for chromosomes 7, 8, and 12 was performed on paraffin- embedded tissue sections and touch imprint preparations of 53 cases of human prostate cancer. Subsequent haematoxylin and eosin (H and E) staining of the hybridized tissue sections allowed unambiguous assignment of hybridization signals either to tumour or to non-tumorous parenchyma. Fifty-three cases of human prostate cancer were evaluated for numerical aberrations of chromosome 7. Scoring 200 cells of tumour and non-tumorous parenchyma in each case revealed abnormalities exclusively in tumour parenchyma in 41 cases (77 per cent). Ten of 41 cases (24 per cent) showed trisomy 7, and 15 cases (37 per cent) monosomy 7 or trisomy 7 in combination with monosomy 7, respectively. Sixteen cases (39 per cent) exhibited polysomy 7 in cells of the tumour parenchyma. In the tumour tissue in one case, different polyploid clones (triploid, tetraploid) and polysomy 7 could be identified by double hybridization with chromosome-specific DNA probes for chromosome 7, plus 8 or 12. The indicated numerical aberrations of chromosome 7 were correlated with 78 per cent of advanced pathological stages or poorly differentiated tumours (pT3/4 or G3) of prostate carcinomas. A statistical analysis of the data revealed significant relationships of particular numerical abnormalities of chromosome 7 to different pathological categories (pT, G, pN) of tumour classification. For the T-classification, the frequency of cells carrying polysomy 7 and polysomy 7/+7 increases significantly from pT1 to pT3/4 (P=0·022). A significant increase from G1 to G3 also became apparent for the total frequencies of numerical abnormal cells (P=0·05).
AU  - Zitzelsberger, H.
AU  - Szücs, S.
AU  - Weier -, H.U.G.
AU  - Lehmann, L.
AU  - Braselmann, H.
AU  - Enders, S.
AU  - Schilling, A.I.
AU  - Breul, J.
AU  - Höfler, H.H.
AU  - Bauchinger, M.
C1  - 40016
C2  - 38061
SP  - 325-335
TI  - Numerical abnormalities of chromosome 7 in human prostate cancer detected by fluorescence in situ hybridization (FISH) on paraffin-embedded tissue sections with centromere-specific DNA probes.
JO  - J. Pathol.
VL  - 172
IS  - 4
PY  - 1994
SN  - 0022-3417
ER  -