Insulin production in the ? cell is controlled primarily at the level of transcription, a process which is regulated through the action of transcription factors specifically expressed in the ? cell and via histone modification of insulin gene chromatin structure. The overall goal of this project will be to further investigate the process whereby insulin gene activation occurs and to define acute and chronic changes that occur in chromatin structure in response to nutrients such as glucose. I hypothesize that transcription of the insulin gene is regulated, in part, through changes in chromatin, which are mediated by Pdx-1, a member of the homeobox class of transcription factors. I further hypothesize that in states of disease, like diabetes mellitus, there are defects at the chromatin level that alter gene expression patterns.
In Aim 1 of this proposal, I will characterize the role of Pdx-1 in glucose-stimulated insulin gene transactivation, using Pdx-1 deletion studies in whole animals and isolated human islets. In this aim, I will establish how Pdx-1 functionally links extracellular glucose levels to insulin transcription, histone modifications, and metabolic control.
Aim 2 will characterize the interactions of Pdx-1 with novel components of the basal transcriptional machinery in the ? cell, concentrating initially on the interaction of Pdx-1 with Paf1, a protein that associates with RNA polymerase II. Other novel interacting proteins will be identified using MALDI-TOF mass spectrometry, and interactions will be verified by mammalian two-hybrid assay. The biological relevance of these interactions in the ? cell will be tested using siRNA and chromatin immunoprecipitation assays.
In Aim 3, I will characterize defects in histone modification and turnover in hyperinsulinemic and hypoinsulinemic rodent models of Type 2 diabetes and human islets chronically exposed to high glucose, using chromatin analysis techniques, such as the micrococcal digest assay and chromatin immunoprecipitation. Relevance: Diabetes mellitus (DM) is a metabolic disease that results from either a complete or relative deficiency of the hormone insulin. DM currently affects 20.8 million people in the U.S. and is increasing in incidence. The cost of caring for this disease is enormous and exceeds over 100 billion dollars annually. In this proposal, the precise mechanisms by which the pancreas produces the hormone insulin will be explored at the level of the gene encoding insulin. Research of this type has the potential to guide the development of new therapies for DM.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Clinical Investigator Award (CIA) (K08)
Project #
5K08DK080225-06
Application #
8116028
Study Section
Diabetes, Endocrinology and Metabolic Diseases B Subcommittee (DDK)
Program Officer
Hyde, James F
Project Start
2007-09-25
Project End
2013-04-30
Budget Start
2011-09-01
Budget End
2013-04-30
Support Year
6
Fiscal Year
2011
Total Cost
$141,443
Indirect Cost
Name
Indiana University-Purdue University at Indianapolis
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
603007902
City
Indianapolis
State
IN
Country
United States
Zip Code
46202
Blue, Emily K; Ballman, Kimberly; Boyle, Frances et al. (2015) Fetal hyperglycemia and a high-fat diet contribute to aberrant glucose tolerance and hematopoiesis in adult rats. Pediatr Res 77:316-25
Kono, Tatsuyoshi M; Sims, Emily K; Moss, Dan R et al. (2014) Human adipose-derived stromal/stem cells protect against STZ-induced hyperglycemia: analysis of hASC-derived paracrine effectors. Stem Cells 32:1831-42
Rajashekhar, Gangaraju; Ramadan, Ahmed; Abburi, Chandrika et al. (2014) Regenerative therapeutic potential of adipose stromal cells in early stage diabetic retinopathy. PLoS One 9:e84671
Erdel, Blake L; Juneja, Rattan; Evans-Molina, Carmella (2014) A case of calciphylaxis in a patient with hypoparathyroidism and normal renal function. Endocr Pract 20:e102-5
Sims, Emily K; Evans-Molina, Carmella (2014) Urinary biomarkers for the early diagnosis of retinopathy and nephropathy in type 1 diabetes mellitus: a ""steady stream"" of information using proteomics. Transl Res 163:183-7
Evans-Molina, C; Hatanaka, M; Mirmira, R G (2013) Lost in translation: endoplasmic reticulum stress and the decline of ?-cell health in diabetes mellitus. Diabetes Obes Metab 15 Suppl 3:159-69
Sims, Emily K; Hatanaka, Masayuki; Morris, David L et al. (2013) Divergent compensatory responses to high-fat diet between C57BL6/J and C57BLKS/J inbred mouse strains. Am J Physiol Endocrinol Metab 305:E1495-511
Chaudhry, Zunaira Z; Morris, David L; Moss, Dan R et al. (2013) Streptozotocin is equally diabetogenic whether administered to fed or fasted mice. Lab Anim 47:257-65
Evans-Molina, Carmella; Mirmira, Raghavendra G (2013) Achieving ""PeaK-A"" insulin secretion. Diabetes 62:1389-90
Kono, Tatsuyoshi; Ahn, Geonyoung; Moss, Dan R et al. (2012) PPAR-? activation restores pancreatic islet SERCA2 levels and prevents ?-cell dysfunction under conditions of hyperglycemic and cytokine stress. Mol Endocrinol 26:257-71

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