The direct effects of tacrolimus and cyclosporin A on isolated human islets: A functional, survival and gene expression study

The use of immunosuppressive drugs in transplanted patients is associated with the development of diabetes, possibly due to β-cell toxicity. To better understand the mechanisms leading to post-transplant diabetes, we investigated the actions of prolonged exposure of isolated human islets to therapeutical levels of tacrolimus (Tac) or cyclosporin A (CsA). Islets were isolated from the pancreas of multiorgan donors by enzymatic digestion and density gradient centrifugation. Functional, survival and molecular studies were then performed after 4 days of incubation with therapeutical concentrations of Tac or CsA. Glucose-induced insulin secretion was significantly decreased in Tac, but not in CsA exposed islets, which was associated with a reduction of the amount of insulin granules as shown by electron microscopy. The percentage of apoptotic β-cells was higher in Tac than CsA exposed islets. Microarray experiments followed by Gene Set Enrichment Analysis revealed that gene expression was more markedly affected upon Tac treatment. In conclusion, Tac and CsA affect features of beta-cell differently, with several changes occurring at the molecular level.

[1]  J. Bradley,et al.  Immunosuppressive agents in solid organ transplantation: Mechanisms of action and therapeutic efficacy. , 2005, Critical reviews in oncology/hematology.

[2]  H. Bonatti.  Risk Factors Associated With the Onset and Progression of Posttransplantation Diabetes in Renal Allograft Recipients , 2008 .

[3]  Terence P. Speed,et al.  Quality Assessment of Affymetrix GeneChip Data , 2005 .

[4]  A. Secchi,et al.  Secretory defects induced by immunosuppressive agents on human pancreatic β-cells , 2002, Acta Diabetologica.

[5]  E. Oetjen,et al.  Inhibition of human insulin gene transcription by the immunosuppressive drugs cyclosporin A and tacrolimus in primary, mature islets of transgenic mice. , 2003, Molecular pharmacology.

[6]  Benjamin M. Bolstad,et al.  affy - analysis of Affymetrix GeneChip data at the probe level , 2004, Bioinform..

[7]  A. Ojo.  Cardiovascular Complications After Renal Transplantation and Their Prevention , 2006, Transplantation.

[8]  Reversibility of the acute toxic effect of Cyclosporin A on pancreatic B cells of Wistar rats , 1986, Diabetologia.

[9]  U. Boggi,et al.  Functional and molecular defects of pancreatic islets in human type 2 diabetes. , 2005, Diabetes.

[10]  P. Marchetti.  New‐onset diabetes after liver transplantation: From pathogenesis to management , 2005, Liver transplantation : official publication of the American Association for the Study of Liver Diseases and the International Liver Transplantation Society.

[11]  V. Ravindran,et al.  The burden of new‐onset diabetes mellitus after transplantation , 2006, Clinical transplantation.

[12]  Robert Gentleman,et al.  Using GOstats to test gene lists for GO term association , 2007, Bioinform..

[13]  K. Kohnert,et al.  Toxic effects of cyclosporine on the endocrine pancreas of Wistar rats. , 1986, Transplantation.

[14]  U. Boggi,et al.  The endoplasmic reticulum in pancreatic beta cells of type 2 diabetes patients , 2007, Diabetologia.

[15]  A. Elayat,et al.  Functional and morphological study of cultured pancreatic islets treated with cyclosporine. , 2007, Life sciences.

[16]  A. Hartmann,et al.  A 6-year prospective study on new onset diabetes mellitus, insulin release and insulin sensitivity in renal transplant recipients. , 2003, Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association.

[17]  A. Gambotto,et al.  Adenovirus-mediated XIAP gene transfer reverses the negative effects of immunosuppressive drugs on insulin secretion and cell viability of isolated human islets. , 2005, Diabetes.

[18]  L. Sower,et al.  Differential effects of interleukin-2 blockade on apoptosis in naïve and activated human lymphocytes12 , 2003, Transplantation.

[19]  Gordon K. Smyth,et al.  limma: Linear Models for Microarray Data , 2005 .

[20]  U. Boggi,et al.  Lipotoxicity in human pancreatic islets and the protective effect of metformin. , 2002, Diabetes.

[21]  A. Penfornis,et al.  Immunosuppressive drug-induced diabetes. , 2006, Diabetes & metabolism.

[22]  Rafael A Irizarry,et al.  Exploration, normalization, and summaries of high density oligonucleotide array probe level data. , 2003, Biostatistics.

[23]  M. Schnitzler,et al.  Guidelines for the treatment and management of new‐onset diabetes after transplantation 1 , 2005, Clinical transplantation.

[24]  M. Márques,et al.  Tacrolimus‐Induced Diabetes in Rats Courses with Suppressed Insulin Gene Expression in Pancreatic Islets , 2007, American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons.

[25]  J. Nerup,et al.  Direct effects of cyclosporin A on human pancreatic beta-cells. , 1986, Diabetes.

[26]  Yuichiro Yamada,et al.  Tacrolimus suppresses glucose-induced insulin release from pancreatic islets by reducing glucokinase activity. , 2005, American journal of physiology. Endocrinology and metabolism.

[27]  J. Neilson,et al.  Calcineurin/NFAT signalling regulates pancreatic beta-cell growth and function. , 2006, Nature.

[28]  R. Bloom,et al.  New-onset diabetes mellitus in the kidney recipient: diagnosis and management strategies. , 2008, Clinical journal of the American Society of Nephrology : CJASN.

[29]  Pablo Tamayo,et al.  Gene set enrichment analysis: A knowledge-based approach for interpreting genome-wide expression profiles , 2005, Proceedings of the National Academy of Sciences of the United States of America.