After an absence from academic research, I chose to return to the University of Florida to be a part of its internationally recognized Diabetes Research Group. As a junior member of this distinguished group, I have access to human and animal samples for parallel studies of the immunopathogenesis of Type 1 diabetes, and the support a large, diverse university's resources for advanced technology, collaboration, and funding. I have a wide breath of technical skills, especially in the design or adapt methodology. However, my further development as a research scientist depends on refining these skills to execute a focused research plan. The writing of this KO1 Career Development Award proposal gives me the opportunity to gain experience in crucial areas where I lack depth: clinical research design, grant project development, and scientific authorship. This proposal addresses fundamental defects in antigen presenting cell (APC) function important in immunopathogenesis of Type 1 Diabetes (IDDM). We have reported a biochemical component of APC dysfunction in diabetic/at-risk humans and in the Nonobese diabetic (NOD) mouse: the aberrant expression of the normally inducible cyclooxygenase, prostaglandin synthase 2 (PGS2/COX2). Sequence analysis indicates this phenotype is not due to a cis defect in the PGS2 structural gene or its immediate 5' or 3' regions. These finding suggest that components further up- or downstream and/or trans acting molecules may be responsible for its aberrant expression. In our analysis of trans acting factors, we found that PGS2 expression in NOD macrophages and in the peripheral blood monocytes of 55% of at- risk/IDDM subjects tested was resistant to IL10 suppression. In addition, the PGS2 expression of these cells could be further enhanced by even a brief exposure to GM-CSF(granulocyte monocyte colony stimulating factor). In contrast, PGS2 expression of control cells is not induced by treatment with GM-CSF and is IL 10 sensitive after LPS (lipopolysaccharide) induction. These aspects of NOD/IDDM PGS2 expression are not due to a defect in IL 10 production or its general functional activity, but appears to be linked to GM-CSF activation of STAT5, a transcription regulator of the PGS2 gene's enhancer. Preliminary analyses indicate a loss of DNA binding of STAT dominant negative repressor (truncated) isoforms and a persistence of binding of its full-length activator forms may contribute to aberrant PGS2 expression seen in NOD and at-risk/IDDM human myeloid cells. Our goal is to examine STAT5 signal transduction/transcription regulation in Type 1 diabetic patients, their at-risk relatives, and in the NOD mouse, and determine its 1) role in monocyte and macrophage PGS2 expression dysregulation, 2) possible effects on myeloid differentiation, and 3) relationship to APC dysfunction in the immunopathogenesis of Type 1 Diabetes.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Scientist Development Award - Research & Training (K01)
Project #
1K01DK002947-01A1
Application #
6400404
Study Section
Diabetes, Endocrinology and Metabolic Diseases B Subcommittee (DDK)
Program Officer
Hyde, James F
Project Start
2001-09-01
Project End
2004-08-31
Budget Start
2001-09-01
Budget End
2002-08-31
Support Year
1
Fiscal Year
2001
Total Cost
$87,738
Indirect Cost
Name
University of Florida
Department
Pathology
Type
Schools of Medicine
DUNS #
073130411
City
Gainesville
State
FL
Country
United States
Zip Code
32611
Garrigan, Erin; Belkin, Nicole S; Seydel, Federica et al. (2015) Csf2 and Ptgs2 Epigenetic Dysregulation in Diabetes-prone Bicongenic B6.NODC11bxC1tb Mice. Genet Epigenet 7:5-17
Garrigan, Erin; Belkin, Nicole S; Alexander, John J et al. (2013) Persistent STAT5 phosphorylation and epigenetic dysregulation of GM-CSF and PGS2/COX2 expression in Type 1 diabetic human monocytes. PLoS One 8:e76919
Seydel, F; Garrigan, E; Stutevoss, B et al. (2008) GM-CSF induces STAT5 binding at epigenetic regulatory sites within the Csf2 promoter of non-obese diabetic (NOD) mouse myeloid cells. J Autoimmun 31:377-84
Rumore-Maton, B; Elf, J; Belkin, N et al. (2008) M-CSF and GM-CSF regulation of STAT5 activation and DNA binding in myeloid cell differentiation is disrupted in nonobese diabetic mice. Clin Dev Immunol 2008:769795
Xiang, Zhimin; Litherland, Sally A; Sorensen, Nicholas B et al. (2006) Pharmacological characterization of 40 human melanocortin-4 receptor polymorphisms with the endogenous proopiomelanocortin-derived agonists and the agouti-related protein (AGRP) antagonist. Biochemistry 45:7277-88
Litherland, Sally A; Grebe, Kristie M; Belkin, Nicole S et al. (2005) Nonobese diabetic mouse congenic analysis reveals chromosome 11 locus contributing to diabetes susceptibility, macrophage STAT5 dysfunction, and granulocyte-macrophage colony-stimulating factor overproduction. J Immunol 175:4561-5
Litherland, S A; Xie, T X; Grebe, K M et al. (2005) Signal transduction activator of transcription 5 (STAT5) dysfunction in autoimmune monocytes and macrophages. J Autoimmun 24:297-310
Litherland, S A; Xie, T X; Grebe, K M et al. (2004) IL10 resistant PGS2 expression in at-risk/Type 1 diabetic human monocytes. J Autoimmun 22:227-33
Lian, Wei; Litherland, Sally A; Badrane, Hassan et al. (2004) Ultrasensitive detection of biomolecules with fluorescent dye-doped nanoparticles. Anal Biochem 334:135-44