The research proposed in this application will develop and utilize massively parallel gene analysis technology. Both commercially available microarrays from Affymetrix and arrays custom-made in this facility will be used. The basic strategy for these experiments is to identify genes that are relevant for individual experimental projects. These finite gene sets will be included on arrays made in our facility, which will be customized for the individual user. The advantages of such an approach are that multiple experiments will then be performed cost effectively, allowing appropriate statistical analyses of data, as well as systematic variations and manipulations. Ongoing studies are those from NIDDK-funded investigators examining disease processes and models in humans, animals and homogenous cells: 1) Nephropathy occurring during the course of experimental diabetes mellitus in mice; 2) Lymphocytic abnormalities in mice with experimental inflammatory bowel disease (IBD); 3) Molecular basis of intestinal epithelial adaptation to altered luminal sodium load; 4) Activation defects in monocytes from patients with IBD; 5) Effects of steroids on muscle and adipocyte gene expression in polycystic ovary syndrome; 6) Analysis of signaling defects in mouse models of thyroid hormone resistance; 7) Analysis of signaling defects in patients with thyroid hormone resistance; 8) Analyses of mRNA changes in human pancreatic islets prepared for transplantation; 9) Effects of high glucose exposure to pancreatic beta cells; and, 10) Neuronal gene expression in response to gonadotropin-releasing hormone. These analyses will permit the identification of relevant genes that are upregulated or downregulated in these various conditions. Pilot and feasibility studies will be to: 1) Develop cDNA microarrays from pituitary, pancreatic islets and renal glomeruli to allow gene profiling from these anatomically unique sites; and 2) Utilize laser capture microdissection microscopy to obtain glomerular RNA from animal models and human tissue specimens of lupus nephritis, and subsequently apply microarray technology to identify gene changes in this disease. If this NIDDK-sponsored core facility is funded, innovative work investigating gene expression in a variety of renal, endocrinologic and gastrointestinal disease models and processes will be performed.
