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4. Molecular mechanisms of toxicity and cell damage in clonal human and mouse pancreatic beta cells, alpha cells, and intestinal L cells exposed to diabetic milieu

Author

Vasu, Srividya

Subjects
616.4
Abstract

Novel human pancreatic beta cell line, 1.IB4 was generated only recently by electrofusion of human islet cells and immortal P ANC-I cell line. In contrast, the mouse insulinoma cell line - MIN6, pancreatic alpha cell line - Alpha TCI clone 9 and GLP-I secreting enteroendocrine cell line - GLUTag are widely used in diabetes research. While cellular responses of pancreatic beta cells on exposure to diabetic conditions have been studied extensively, responses of this novel human insulin secreting cell line and pancreatic alpha and intestinal L cells have not been evaluated previously. In this thesis, mechanisms of toxicity and cell damage in 1.1B4, MIN6, Alpha TCI clone 9 and GLUTag cells exposed t9 diabetic milieu were evaluated. 1.1B4, MIN6 and GLUTag cells suffered cell death on exposure to chemicals, high glucose, palmitate and cytokines while Alpha TCI clone 9 cells were relatively resistant to these agents. 1.lB4 and MIN6 cell secretory function, insulin content and mRNA expression of genes involved in stimulus secretion coupling were affected to differing degrees. Expression of antioxidant enzyme genes was upregulated but was insufficient to detoxify ROS generated by toxins with the exception of Alpha TCI clone 9 cells. Pro-inflammatory cytokines caused nitrosative stress while other toxins caused oxidative stress, which was evident from the increase in DNA fragmentation and numbers of apoptotic cells. Mechanisms of toxicity and cell damage in the beta cell lines, 1.lB4 and MIN6, were consistent with previously established results using rodent cell models and isolated islets. This is the first study to report cellular responses of 1.1B4, GLUTag and Alpha TCI clone 9 cells under diabetic conditions The results indicate that 1.1 B4 cells represent a good model of human islet beta cells for studying beta cell biology. Further, whereas MIN6 and GLUTag cells are sensitive to cytotoxic damage, Alpha Tel clone 9 cells exhibit resistance due to their relatively higher expression of catalase and lower expression of Glut2. Further dissection of molecular pathways could aid in better understanding of mechanisms involved in diabetes, leading to new therapeutic approaches.

Published
2012

9. Circulating miRNA in Patients Undergoing Total Pancreatectomy and Islet Autotransplantation

Author

Vasu, Srividya, primary, Yang, Jiemin M., additional, Hodges, James, additional, Abu-El-Haija, Maisam A., additional, Adams, David B., additional, Balamurugan, Appakalai N., additional, Beilman, Greg J., additional, Chinnakotla, Srinath, additional, Conwell, Darwin L., additional, Freeman, Martin L., additional, Gardner, Timothy B., additional, Hatipoglu, Betul, additional, Kirchner, Varvara, additional, Lara, Luis F., additional, Morgan, Katherine A., additional, Nathan, Jaimie D., additional, Posselt, Andrew, additional, Pruett, Timothy L., additional, Schwarzenberg, Sarah J., additional, Singh, Vikesh K., additional, Wijkstrom, Martin, additional, Witkowski, Piotr, additional, Naziruddin, Bashoo, additional, and Bellin, Melena D., additional

Published
2021
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17. Beneficial effects of parenteral GLP-1 delivery by cell therapy in insulin-deficient streptozotocin diabetic mice.

Author

Vasu, Srividya, Moffett, R. C., McCluskey, J. T., Hamid, Muhajir, Irwin, N., Flatt, P. R., Vasu, Srividya, Moffett, R. C., McCluskey, J. T., Hamid, Muhajir, Irwin, N., and Flatt, P. R.

Abstract

Parenteral delivery of long-Acting glucagon-like peptide-1 (GLP-1) mimetics has received much attention as a therapeutic option for diabetes. However, cell therapy-based GLP-1 treatments may provide a more physiological regulation of blood glucose. The present study assessed the effects of chronic GLP-1 delivery by cell therapy, using the GLP-1-secreting GLUTag cell line, in normoglycemic and streptozotocin-induced diabetic mice. GLUTag cell aggregates were transplanted into the subscapular region of mice. Over 30 days, cellular transplantation gave rise to encapsulated and well-vascularized growths, which contained immunoreactive GLP-1. Cell implantation was well tolerated and had no appreciable metabolic effects in normal mice. However, transplantation significantly (P<0.001) countered excessive food and fluid intake in diabetic mice and maintained normal body weight. Circulating glucose (P<0.01) and glucagon (P<0.05) were significantly reduced and plasma insulin and GLP-1 dramatically increased. This was associated with significantly (P<0.01) improved glucose tolerance in diabetic mice. Histological examination of the pancreata of these mice revealed elevations (P<0.001) in islet and β-cell area, with reduced (P<0.001) -cell area. Increased β-cell mass reflected the enhanced proliferation relative to apoptosis. These studies emphasize the potential of chronic GLP-1 delivery by cell therapy as a potential therapeutic option for diabetes.

Published
2013

19. Responses of GLPI-secreting L-cells to cytotoxicity resemble pancreatic ß-cells but not α-cells.

Author

Vasu, Srividya, Moffett, R. Charlotte, McClenaghan, Neville H., and Flatt, Peter R.

Subjects
*GLUCAGON-like peptide 1, *CYTOKINES, *STREPTOZOTOCIN, *DIABETES, *CELL proliferation, *CELL-mediated cytotoxicity, *ANTIOXIDANTS, *LABORATORY mice
Abstract

Little is known about responses of intestinal L-cells to chemical or cytokine-mediated attack and how these compare with pancreatic ß- or α-cells. Administration of streptozotocin to mice induced severe diabetes, islet lymphocytic infiltration, increased α-cell proliferation and decreased numbers of ß- and L-cells. In vitro, streptozotocin and cytokines reduced cell viability with higher lethal dose 50 values for α-TC1 cells. mRNA expression of Glut2 was lower and Cat was greater in GLUTag and α-TC1 cells compared with MIN6 cells. Cytotoxins affected the transcription of genes involved in secretion in GLUTag and MIN6 cells. They are also involved in upregulation of antioxidant defence enzymes, transcription of Nf?B and Nos2, and production of nitrite in all cell types. Cytotoxin-induced DNA damage and apoptosis were apparent in all cells, but α-TC1 cells were less severely affected. Thus, responses of GLP1-secreting L-cells to cytotoxicity resemble ß-cells, whereas a-cells are resistant due to differences in the expression of genes involved in cytotoxicity or antioxidant defence. [ABSTRACT FROM AUTHOR]

Published
2015
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20. Acknowledgment to Reviewers of the International Journal of Molecular Sciences in 2020.

Subjects
QUALITY standards, GRATITUDE
Abstract

Peer review is the driving force of journal development, and reviewers are gatekeepers who ensure that the I International Journal of Molecular Sciences i maintains its standards for the high quality of its published papers. Thanks to the cooperation of our reviewers, in 2020, the median time to first decision was 15 days and the median time to publication was 33 days. [Extracted from the article]

Published
2021
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