Alternative RNA splicing events contribute to the onset of islet dysfunction in type 1 diabetes
EMSL Project ID
60072
Abstract
Type 1 diabetes (T1D) is a results from the autoimmune destruction of pancreatic β cells, which are the cells responsible for insulin production and secretion in the boby. Despite T1D is primarily associated with autoimmune response, recent studies have found numeros genes expressed in pancreatic β cells that are associated with the disease development. Studies from our group and others have provided evidence of a variety of proteins that are differentially processed by splicing during T1D development. Therefore, we hypothesize that altered RNA splicing events contribute to β cell dysfunction, potentially by forming of immunogenic protein isoforms to trigger autoimmune response. The next generation sequencing has immensely improve genome wide RNA analysis, which has revealed the complexity of the transcriptome providing insights on the physiological and pathological processes. This includes a growing appreciation for the role of alternative splicing in many biological processes. In fact, there is a large number of human diseases caused by mutations in the RNA splicing machinery. In the pancreas, a considerable dysregulation of RNA binding proteins (RBPs) and aberrant mRNA splicing has been shown in islets from donor with type 2 diabetic patients compared to healthy controls. Despite that splicing network in pancreas has been studied, only limited information is available in the context of T1D. In fact, an important knowledge gap is what are processes and consequences of alternative splicing regulation during the T1D development. We propose to investigate the role of alternative splicing in β cells in processes relevant to T1D development. Our goal is to understand the how anternative splicing contribute to the autoimmune response and β cell death. To accomplish these goals, we propose the following specific aims: Aim 1. Use novel RNA sequencing technologies to identify conserved alternatively spliced transcripts in mouse and human T1D islet samples; Aim 2. Determine the localization of candidate RNA isoforms in individual cells in intact human islets using fliFISH as a high accuracy smFISH approach that localizes individual transcripts with 20-30 nm resolution using STORM microscopy; Aim 3. Use advanced proteomics and innovative computational approaches to identify candidate novel protein products generated from alternative spliced transcripts in T1D β cells; and Aim 4. Determine the functional ramifications of altered splicing events in β cells by deleting a splicing factor important for regulating b cell function and survival.
Project Details
Start Date
2021-06-15
End Date
2023-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
Co-Investigator(s)