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Gene profiling characterises the patho-physiological sequence of chronic allograft nephropathy (CAN). Effects of isotretinoin (IT)
296
Symposium: Transplantation - Oral Presentations / Pathology - Research and Practice 200 (2004) 296-298
Methods: Pretreatment paraffin-embedded endoscopic esophageal tumor biopsies of 38 patients with advanced (uT3N1) esophageal adenocarcinoma (Barrett's carcinoma) were included. The patients underwent two cycles of cisplatin and fluorouracil (5-FU) therapy with or without additional paclitaxel (taxol) followed by abdominothoracal esophagectomy. RNA expression levels of 5-FU associated genes (TS, TP, DPD, MTHFR, E - M A P l l 5 , ELF3) as well as of platinum and taxane associated genes (ERCC1, ERCC4, HER2-neu, Caldesmon, GADD45, MRP1) were determined using real-time PCR. Expression levels were correlated with the histopathological response to chemotherapy as assessed in surgically resected esophagectomy specimens. Results: High expression levels of MTHFR, ERCC4, Caldesmon and MRP1 were significantly associated with histopathologically determined complete or subtotal tumor regression (p = 0.001; positive predictive value >90% for MTHFR and MRP1 and p < 0.02; positive predictive values >80% for ERCC4 and Caldesmon). High expression levels of DP and TP also showed a strong but statistically not significant correlation with complete or subtotal tumor response. Conclusion: Our results indicate that tumor profiling of mRNA expression of selected genes may be used for the prediction of the prognosis of individual cancer patients with chemotherapentically treated advanced adenocarcinoma of the esophagus
5ymp i : Transp!anta{io n 155
Potential risk factors for chronic allografl nephropathy revealed from protocoll biopsies after renal transplantation M. MENGEL l , W. GWINNER 2, J. RADERMACHER 2, H. HISS 2, H. HALLER 2 , H. KREIPE ~, A. SCHWARZ2 1Institut ftir Pathologie 2Abteilung ftir Nephrologie Medizinische Hochschule Hannover
Aims: Chronic allograft nephropathy is the main cause of graft failure and might be detected on biopsy before graft dysfunction becomes clinically evident. Protocol biopsies, taken during periods of stable graft function, constitute a meaningful strategy to reveal early surrogate markers of unfavourable long-term graft function facilitating a timely therapeutically intervention. Methods: At the Hannover transplant centre for all renal allograft recipients three protocol biopsies (pBx) are aspired at 6, 12, and 26 weeks post transplantation. Between 10/2000 and 3/2003 299 patients entered the program, providing 688 pBx. In 190 patients all three pBx were available. Biopsies were evaluated following the updated Banff criteria. Morphological findings and clinical parameters (n = 18) were statistically correlated to the onset of chronic allograft nephropathy (CAN = interstitial fibrosis in >5% of cortical area and tubular atrophy). Results: 37% (70/190) of patients with three pBx developed signs of CAN within 26 weeks after transplantation. These patients had statistically significant lower calculated glomerular filtration rate (GFR) at all three pBx, more often brain death cadaver than living donors, longer cold ischemia time, more episodes of acute rejection, and more intimal fibrosis in their arteries. A negative correlation was found between the degree of CAN and the GFR at the time of all pBx. In patients with CAN and reduced GFR at pBx 3, the GFR was already reduced at pBx 1, independently from morphological signs of CAN at this time point.
Conclusions: Reduction of GFR precedes manifestation of morphological signs of CAN in protocol biopsies. The functional impairment might represent a reversible stage of allograft injury, allowing for timely therapeutically intervention.
156
Gene profiling characterises the patho-physiological seqence of chronic allograft nephropathy (CAN). Effects of isotretinoin (IT) E. KISS 1, M. HERGENHAHN 2, J. ADAMS 3, S. WANG 1, P. NELSON 4, H.-J. GRONE 1 t Dept. of Molecular and Cellular Pathology and 2Dept. Genetic Alterations and Carcinogenesis German Cancer Research Cnter, Heidelberg 3Dept. of Urology, Karl Ruprecht University, Heidelberg 4Medical Policlinic, University of Munich, Munich
Aims: We have reported a model of CAN in F344 to Lew renal transplantation (TX) characterized by development of early chronic changes (at day 14 post TX) and progressing to CAN after 56 days. Beneficial effects of IT on rejection phenomena in this model were observed. The underlying molecular mechanisms are not understood. Our purpose was to analyze the gene expression profile of the transition period from acute to chronic rejection and the influence of IT treatment. Methods: Allografts were evaluated at days 7, 14 and 56-day with CAN-. The effect of IT on gene expression was analyzed after 14 days treatment. Oligonucleotide microarrays were used. Real time RT-PCR and immunohistochemistry were performed as confirmatory tools. Results: A significant upregulation of genes of innate and adaptive immunity was observed between days 7 and 14 with some genes showing a further increase in expression between days 14 and 56. IFN-7 and IL-10 were highly expressed at all time points, suggesting involvement of both Thl and Th2 immune responses. Antioxidant enzymes showed no compensatory upregulation but interestingly downregulation at each time point. Treatment with IT for 14 days significantly reduced expression of genes of innate immunity, markers of T cell activation (CD8), cell proliferation, and leukocyte recruitment (VCAM 1). Conclusions: Genes expressed early on in allograft rejection determine the development of CAN. IT prevents CAN primarily by its influence on immune regulatory processes.
157
Expression of T N F - ~ , C M V and MiB1 in heart biopsies with and without heart transplant rejection A. M. M(J-LLER, A. HAHN, K.-M. M()LLER t Institute of Pathology, University Clinic Bergmannsheil, Ruhr-University Bochum
Aims: Up to now, diagnosis and consecutive treatment of heart transplant rejection is based upon endomyocardial biopsies. The negative histomorphological diagnosis can not exclude a forthcoming transplant rejection. We studied whether additional immunohistochemical investigations provide foreboding signs of transplant rejection, thereby enabeling the pathologist to counsel the clinician concerning a repeated biopsy. Materials and Methods: 122 endomyocardial biopsies (33 biopsies with rejection grade 1 according to ISHLT, 89 biopsies with grade 0) and - as control - 26 biopsies taken for diagnostic reasons from patients with cardiomyopathy respectively arterial hyperten-
Symposium: Transplantation - Oral Presentations / Pathology - Research and Practice 200 (2004) 296-298
Methods: Pretreatment paraffin-embedded endoscopic esophageal tumor biopsies of 38 patients with advanced (uT3N1) esophageal adenocarcinoma (Barrett's carcinoma) were included. The patients underwent two cycles of cisplatin and fluorouracil (5-FU) therapy with or without additional paclitaxel (taxol) followed by abdominothoracal esophagectomy. RNA expression levels of 5-FU associated genes (TS, TP, DPD, MTHFR, E - M A P l l 5 , ELF3) as well as of platinum and taxane associated genes (ERCC1, ERCC4, HER2-neu, Caldesmon, GADD45, MRP1) were determined using real-time PCR. Expression levels were correlated with the histopathological response to chemotherapy as assessed in surgically resected esophagectomy specimens. Results: High expression levels of MTHFR, ERCC4, Caldesmon and MRP1 were significantly associated with histopathologically determined complete or subtotal tumor regression (p = 0.001; positive predictive value >90% for MTHFR and MRP1 and p < 0.02; positive predictive values >80% for ERCC4 and Caldesmon). High expression levels of DP and TP also showed a strong but statistically not significant correlation with complete or subtotal tumor response. Conclusion: Our results indicate that tumor profiling of mRNA expression of selected genes may be used for the prediction of the prognosis of individual cancer patients with chemotherapentically treated advanced adenocarcinoma of the esophagus
5ymp i : Transp!anta{io n 155
Potential risk factors for chronic allografl nephropathy revealed from protocoll biopsies after renal transplantation M. MENGEL l , W. GWINNER 2, J. RADERMACHER 2, H. HISS 2, H. HALLER 2 , H. KREIPE ~, A. SCHWARZ2 1Institut ftir Pathologie 2Abteilung ftir Nephrologie Medizinische Hochschule Hannover
Aims: Chronic allograft nephropathy is the main cause of graft failure and might be detected on biopsy before graft dysfunction becomes clinically evident. Protocol biopsies, taken during periods of stable graft function, constitute a meaningful strategy to reveal early surrogate markers of unfavourable long-term graft function facilitating a timely therapeutically intervention. Methods: At the Hannover transplant centre for all renal allograft recipients three protocol biopsies (pBx) are aspired at 6, 12, and 26 weeks post transplantation. Between 10/2000 and 3/2003 299 patients entered the program, providing 688 pBx. In 190 patients all three pBx were available. Biopsies were evaluated following the updated Banff criteria. Morphological findings and clinical parameters (n = 18) were statistically correlated to the onset of chronic allograft nephropathy (CAN = interstitial fibrosis in >5% of cortical area and tubular atrophy). Results: 37% (70/190) of patients with three pBx developed signs of CAN within 26 weeks after transplantation. These patients had statistically significant lower calculated glomerular filtration rate (GFR) at all three pBx, more often brain death cadaver than living donors, longer cold ischemia time, more episodes of acute rejection, and more intimal fibrosis in their arteries. A negative correlation was found between the degree of CAN and the GFR at the time of all pBx. In patients with CAN and reduced GFR at pBx 3, the GFR was already reduced at pBx 1, independently from morphological signs of CAN at this time point.
Conclusions: Reduction of GFR precedes manifestation of morphological signs of CAN in protocol biopsies. The functional impairment might represent a reversible stage of allograft injury, allowing for timely therapeutically intervention.
156
Gene profiling characterises the patho-physiological seqence of chronic allograft nephropathy (CAN). Effects of isotretinoin (IT) E. KISS 1, M. HERGENHAHN 2, J. ADAMS 3, S. WANG 1, P. NELSON 4, H.-J. GRONE 1 t Dept. of Molecular and Cellular Pathology and 2Dept. Genetic Alterations and Carcinogenesis German Cancer Research Cnter, Heidelberg 3Dept. of Urology, Karl Ruprecht University, Heidelberg 4Medical Policlinic, University of Munich, Munich
Aims: We have reported a model of CAN in F344 to Lew renal transplantation (TX) characterized by development of early chronic changes (at day 14 post TX) and progressing to CAN after 56 days. Beneficial effects of IT on rejection phenomena in this model were observed. The underlying molecular mechanisms are not understood. Our purpose was to analyze the gene expression profile of the transition period from acute to chronic rejection and the influence of IT treatment. Methods: Allografts were evaluated at days 7, 14 and 56-day with CAN-. The effect of IT on gene expression was analyzed after 14 days treatment. Oligonucleotide microarrays were used. Real time RT-PCR and immunohistochemistry were performed as confirmatory tools. Results: A significant upregulation of genes of innate and adaptive immunity was observed between days 7 and 14 with some genes showing a further increase in expression between days 14 and 56. IFN-7 and IL-10 were highly expressed at all time points, suggesting involvement of both Thl and Th2 immune responses. Antioxidant enzymes showed no compensatory upregulation but interestingly downregulation at each time point. Treatment with IT for 14 days significantly reduced expression of genes of innate immunity, markers of T cell activation (CD8), cell proliferation, and leukocyte recruitment (VCAM 1). Conclusions: Genes expressed early on in allograft rejection determine the development of CAN. IT prevents CAN primarily by its influence on immune regulatory processes.
157
Expression of T N F - ~ , C M V and MiB1 in heart biopsies with and without heart transplant rejection A. M. M(J-LLER, A. HAHN, K.-M. M()LLER t Institute of Pathology, University Clinic Bergmannsheil, Ruhr-University Bochum
Aims: Up to now, diagnosis and consecutive treatment of heart transplant rejection is based upon endomyocardial biopsies. The negative histomorphological diagnosis can not exclude a forthcoming transplant rejection. We studied whether additional immunohistochemical investigations provide foreboding signs of transplant rejection, thereby enabeling the pathologist to counsel the clinician concerning a repeated biopsy. Materials and Methods: 122 endomyocardial biopsies (33 biopsies with rejection grade 1 according to ISHLT, 89 biopsies with grade 0) and - as control - 26 biopsies taken for diagnostic reasons from patients with cardiomyopathy respectively arterial hyperten-