A decline in functional ?-cell mass and subsequent inability to maintain adequate glycemic control are hallmarks of both type 1 and type 2 diabetes. Innovative therapeutic approaches are aimed at preserving and restoring functional ?-cell mass in diabetes; however, strategies to safely expand ?-cell mass remain to be identified. The predominant mechanism for adapting ?-cell mass to states of increased insulin demand is through modulation of ?-cell replication. Therefore, there has been considerable interest in understanding the mechanisms that regulate ?-cell replication with the goal of discovering new therapeutic targets to promote ?-cell regeneration. Preliminary unpublished evidence from our laboratory suggests that the NAD+-dependent cytoplasmic deacetylase Sirtuin 2 (SIRT2) acts as a nutrient-dependent regulator of mitogenic signaling in rodent and human ?-cells. Using mouse genetic and inhibitor approaches in human islets, we found that loss of SIRT2 activity stimulates ?-cell proliferation and ?-cell mass expansion under hyperglycemic conditions. We have also obtained evidence that mimicking nutrient state changes by manipulating NAD+ availability regulates ?-cell proliferation in a manner consistent with SIRT2-dependent responses. Since intracellular NAD+ levels fluctuate with glucose availability, we hypothesize that SIRT2 couples ?-cell proliferation to glucose metabolism. Furthermore, we have found that SIRT2 inhibits ?-cell proliferation by dampening MAPK signaling and that SIRT2 inhibition in systemic hyperglycemia promotes ?-cell proliferation, while protecting ?-cells from activating pro-apoptotic signaling downstream of the endoplasmic reticulum (ER) stress response. In this proposal, we will explore how SIRT2 regulates mitogenic signaling as well as ER stress responses in ?-cells. To accomplish this, we will pursue three Aims.
In Aim 1 we will employ mouse genetic approaches and experiments in human islets to determine how glucose and nutrient state affect SIRT2-dependent regulation of ?-cell proliferation. Here, we will investigate links between NAD metabolism, activity of the master regulator of cellular energy homeostasis AMPK, SIRT2 activity, and ?-cell proliferation to gain mechanistic insight into the signaling cascades that couple nutrient availability to proliferation in ?-cells. To understand how SIRT2 modulates intracellular signaling to affect glucose-induced proliferative and apoptotic responses in ?-cells, in Aim 2, we will identify the downstream effectors of SIRT2 in the regulation of ?-cell proliferation, employing proteomic as well as in vitro and in vivo approaches. Finally, in Aim 3, we will examine the effects of SIRT2 inhibition on human ?-cell proliferation and function in vivo and explore whether combinatorial targeting of different mitogenic signaling pathways can augment pro-proliferative effects of SIRT2 inhibition. Together, experiments under this proposal will uncover how ?-cells translate nutrient cues into mitogenic signals as well as pave the way for developing pharmacological strategies to safely increase ?-cell mass in humans with diabetes.

Public Health Relevance

Nutritional cues have prominent effects on ?-cell proliferation; however, it is still unclear how ?-cells translate nutrient cues into mitogenic signals. Here, we will explore how the cytoplasmic deacetylase SIRT2 regulates mitogenic signaling and cell death responses in ?-cells. Our proposal will define signaling cascades that couple nutrient availability to cell proliferation in ?-cells, as well as pave the way for developing pharmacological strategies to safely increase ?-cell mass in humans with diabetes.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
2R01DK078803-11
Application #
9818751
Study Section
Cellular Aspects of Diabetes and Obesity Study Section (CADO)
Program Officer
Sato, Sheryl M
Project Start
2007-09-01
Project End
2024-06-30
Budget Start
2020-07-01
Budget End
2021-06-30
Support Year
11
Fiscal Year
2020
Total Cost
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
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