The shortage of pancreatic beta-cells exhibiting glucose-responsive insulin secretion has been a major obstacle to the development of widely applicable cell transplantation therapies for diabetes. To address this problem, the ultimate goal of our research is to develop expanded populations of human beta cells or beta-cell precursors. This would provide a large quantity of cells, grown in vitro, that retain glucose- responsive insulin secretion or the ability to differentiate and acquire this property. Such cells could then be used for cell transplantation therapies for diabetes. Two approaches are being pursued to expand human beta-cells and endocrine cell precursors in vitro. The first is to grow primary cells on extracellular matrix (ECM) in the presence of hepatocyte growth factor/scatter factor (HGF/SF). The second is to express dominant oncogenes in the cells, resulting in matrix and growth factor independent growth in vitro. Following the expansion of primary cells, they must be removed from the ECM and aggregated into islet-like cell clusters (ICCs). However, detachment of the expanded primary cells from the matrix results in a form of apoptosis known as anoikis. For the beta-cell line expressing the SV40 T antigen and H-ras Val12 dominant oncogenes, removal of the oncogenes using the cre-lox recombinase system, prior to aggregation into ICCs, results in rapid and efficient apoptosis. Thus, apoptosis has been encountered in two distinct situations in attempts to expand human beta cells and beta-cell precursors in culture. In this proposal, approaches to inhibition apoptosis using inhibitors of nitric oxide and adenoviral vectors expressing anti-apoptotic genes will be explored. The effects of the anti-apoptotic genes on transplantation of primary, non-expanded cultures of human pancreatic endocrine cells will also be studied. These studies are important to the development of expanded populations of human beta cells and beta-cell precursors for cell transplantation therapies for diabetes.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK055065-03
Application #
6178060
Study Section
Special Emphasis Panel (ZDK1-GRB-B (O1))
Program Officer
Sato, Sheryl M
Project Start
1998-09-30
Project End
2002-09-29
Budget Start
2000-09-30
Budget End
2002-09-29
Support Year
3
Fiscal Year
2000
Total Cost
$264,687
Indirect Cost
Name
University of California San Diego
Department
Pediatrics
Type
Schools of Medicine
DUNS #
804355790
City
La Jolla
State
CA
Country
United States
Zip Code
92093
Itkin-Ansari, P; Marcora, E; Geron, I et al. (2005) NeuroD1 in the endocrine pancreas: localization and dual function as an activator and repressor. Dev Dyn 233:946-53
Itkin-Ansari, Pamela; Geron, Ifat; Hao, Ergeng et al. (2003) Cell-based therapies for diabetes: progress towards a transplantable human beta cell line. Ann N Y Acad Sci 1005:138-47
Demeterco, Carla; Itkin-Ansari, Pamela; Tyrberg, Bjorn et al. (2002) c-Myc controls proliferation versus differentiation in human pancreatic endocrine cells. J Clin Endocrinol Metab 87:3475-85
Wood, L D; Halvorsen, T L; Dhar, S et al. (2001) Characterization of ataxia telangiectasia fibroblasts with extended life-span through telomerase expression. Oncogene 20:278-88
de la Tour, D; Halvorsen, T; Demeterco, C et al. (2001) Beta-cell differentiation from a human pancreatic cell line in vitro and in vivo. Mol Endocrinol 15:476-83
Halvorsen, T; Levine, F (2001) Diabetes mellitus-cell transplantation and gene therapy approaches. Curr Mol Med 1:273-86
Demeterco, C; Levine, F (2001) Gene therapy for diabetes. Front Biosci 6:D175-91
Halvorsen, T L; Beattie, G M; Lopez, A D et al. (2000) Accelerated telomere shortening and senescence in human pancreatic islet cells stimulated to divide in vitro. J Endocrinol 166:103-9
Beattie, G M; Leibowitz, G; Lopez, A D et al. (2000) Protection from cell death in cultured human fetal pancreatic cells. Cell Transplant 9:431-8
Itkin-Ansari, P; Demeterco, C; Bossie, S et al. (2000) PDX-1 and cell-cell contact act in synergy to promote delta-cell development in a human pancreatic endocrine precursor cell line. Mol Endocrinol 14:814-22