Islet amyloid formation is a pathological hallmark of type 2 diabetes and is observed in >90% of patients with the disease. These amyloid deposits are associated with -cell apoptosis resulting in -cell mass loss. In the autoimmune disease process of type 1 diabetes, cytokines are responsible for the induction of -cell apoptosis. Studies of the mechanisms of -cell loss have identified the involvement of a number of pro- apoptotic molecules. However, the role of apoptosis repressor with a caspase recruitment domain (ARC), a novel anti-apoptotic protein recently identified in the cell, remains largely unknown. To gai greater insight into the mechanisms of -cell apoptosis, we will determine how ARC interacts with other proteins in the cell to limit apoptosis. These interactions include (a) synergizing wth the anti-apoptotic protein 14-3-3?, (b) inhibiting signaling by the pro-apoptotic protein JNK, an (c) inhibiing signaling pathway molecules activated by TNF-?-, IL-1, and IFN-?. In this application we propose four specific aims to examine the mechanisms by which ARC decreases - cell apoptosis.
Specific Aims 1 -3 will utilize in vitro approaches. The first two aims primaril address islet amyloid-induced mechanisms, while the third focuses on those induced by cytokines.
Specific Aim 4 will be done in vivo to examine the translational relevance of increasing ARC in the cell.
Specific Aim 1 : To determine if ARC's inhibition of JNK and/or p53 is sufficient to prevent amyloid-induced - cell apoptosis.
Specific Aim 2 : To determine the role of 14-3-3? in mediating the effect of ARC to reduce amyloid-induced - cell apoptosis.
Specific Aim 3 : To determine whether ARC prevents cytokine-induced -cell apoptosis and the mechanism(s) by which it does so.
Specific Aim 4 : To determine in vivo whether increasing ARC expression in cells can safely reduce amyloid- induced -cell loss. For the in vitro studies (Specific Aims 1-3) we will use islets from our transgenic mouse model of islet amyloid, wild type mice and humans as well as an immortalized -cell line. For the in vivo work examining the effects of ARC in cells (Specific Aim 4), we will produce a new transgenic mouse model expressing both human islet amyloid polypeptide (hIAPP) and ARC in its cells. The long-term goal of this work is to define additional targets to prevent or curtail -cell loss, an important component of the diabetes disease process.

Public Health Relevance

The number of individuals affected by diabetes continues to increase, with the disease disproportionately affecting Veterans. The islet cell represents a key component in the disease process, manifest as reduced insulin secretion and the loss of insulin producing cells. There are a number of causes for the loss of cells, including islet amyloid and cytokines The formation of amyloid deposits and the action of cytokines are associated with cellular stress that activates pathways leading to the death of the cell. We propose to examine proteins in the islet cell that act on molecules in these pathways and thus could prevent -cell death. By gaining a greater understanding of the role of proteins linked to the death and survival of the cell, our goal is ultimately to develop new approaches to treat patients with diabetes in order to preserve their cells.

Agency
National Institute of Health (NIH)
Institute
Veterans Affairs (VA)
Type
Non-HHS Research Projects (I01)
Project #
5I01BX001060-08
Application #
9636447
Study Section
Endocriniology A (ENDA)
Project Start
2010-10-01
Project End
2019-12-31
Budget Start
2019-01-01
Budget End
2019-12-31
Support Year
8
Fiscal Year
2019
Total Cost
Indirect Cost
Name
VA Puget Sound Healthcare System
Department
Type
DUNS #
020232971
City
Seattle
State
WA
Country
United States
Zip Code
98108
Templin, Andrew T; Meier, Daniel T; Willard, Joshua R et al. (2018) Use of the PET ligand florbetapir for in vivo imaging of pancreatic islet amyloid deposits in hIAPP transgenic mice. Diabetologia 61:2215-2224
Salunkhe, Vishal A; Veluthakal, Rajakrishnan; Kahn, Steven E et al. (2018) Novel approaches to restore beta cell function in prediabetes and type 2 diabetes. Diabetologia 61:1895-1901
Morton, Gregory J; Muta, Kenjiro; Kaiyala, Karl J et al. (2017) Evidence That the Sympathetic Nervous System Elicits Rapid, Coordinated, and Reciprocal Adjustments of Insulin Secretion and Insulin Sensitivity During Cold Exposure. Diabetes 66:823-834
de Boer, Ian H; Kahn, Steven E (2017) SGLT2 Inhibitors-Sweet Success for Diabetic Kidney Disease? J Am Soc Nephrol 28:7-10
Templin, Andrew T; Samarasekera, Tanya; Meier, Daniel T et al. (2017) Apoptosis Repressor With Caspase Recruitment Domain Ameliorates Amyloid-Induced ?-Cell Apoptosis and JNK Pathway Activation. Diabetes 66:2636-2645
Hogan, Meghan F; Meier, Daniel T; Zraika, Sakeneh et al. (2016) Inhibition of Insulin-Degrading Enzyme Does Not Increase Islet Amyloid Deposition in Vitro. Endocrinology 157:3462-8
Meier, Daniel T; Entrup, Leon; Templin, Andrew T et al. (2016) The S20G substitution in hIAPP is more amyloidogenic and cytotoxic than wild-type hIAPP in mouse islets. Diabetologia 59:2166-71
Meier, Daniel T; Entrup, Leon; Templin, Andrew T et al. (2015) Determination of Optimal Sample Size for Quantification of ?-Cell Area, Amyloid Area and ?-Cell Apoptosis in Isolated Islets. J Histochem Cytochem 63:663-73
Meier, Daniel T; Tu, Ling-Hsien; Zraika, Sakeneh et al. (2015) Matrix Metalloproteinase-9 Protects Islets from Amyloid-induced Toxicity. J Biol Chem 290:30475-85
Meier, Daniel T; Morcos, Mary; Samarasekera, Thanya et al. (2014) Islet amyloid formation is an important determinant for inducing islet inflammation in high-fat-fed human IAPP transgenic mice. Diabetologia 57:1884-8