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Genomic Characterization of Diabetic Gastroparesis
To date, little is known in regards to the genetic alterations which accompany the onset of gastroparesis (GP) which affects 30-65% of diabetic patients. The goal of this study is to characterize unique changes in RNA expression in gastric tissue and serum of diabetic, gastroparetic patients to identify the molecular pathways that lead to gastric dysfunction. To do this, we have been approved to use serum and gastric tissue samples collected by the Gastroparesis Clinical Research Consortium (GpCRC) on which we propose to isolate mRNA (from gastric tissues) and miRNA (from serum and gastric tissues). These serum and tissue samples are from symptomatic diabetics, who have documented delayed emptying of the stomach, have glucose control with HA1c < 10, no renal impairment (creatinine <2.0 mg/dl) and no active malignancy. Our control group will include diabetic patients who have no symptoms of gastroparesis, have no peripheral neuropathy, have good glucose (HbA1c<10), creatinine <2mg/dl and no malignancy. Once isolated and hybridized to expression arrays, these RNA samples will help to identify genes uniquely expressed or absent in the experimental (diabetic, gastroparetic) vs. control (diabetic-only) group. Briefly we will isolate circulating miRNA from serum-exosomes or mRNA and miRNA from paraffin-embedded gastric tissue samples using commercially available and highly optimized kits. Once isolated, the RNA will be assayed for integrity, and if suitable, amplified, and properly labeled for hybridization onto either whole genome microarray or microRNA expression arrays. Hybridized signal intensities will be assayed and analyzed for statistical significance using appropriate microarray software. Upon completion of this project, we expect to identify clusters of mRNA and microRNA changes on which we can apply pathway analysis software to identify putative pathways that regulate or are regulated by these genes. If awarded, this very sophisticated genetic research project could be recognized as a scientific bridge between two Consortiums, GPCRC and DCC focusing on the same population of patients and their clinical complications.

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