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The role of SRSF1 in liver function and NASH

SRSF1在肝功能和NASH中的作用

基本信息

批准号：
9124530
负责人：
Waqar Arif
金额：
$ 4.86万
依托单位：
UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-05-16 至 2021-05-15
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9124530
关键词：
5&apos Splice Site Ablation Affect Alcohol consumption Alternative Splicing American Arginine Binding Bioinformatics Biological Assay Biological Models Cell Death Cell physiology Cells Computer Analysis Cytoplasm Data Defect Dependovirus Development Dipeptides Disease Disease Progression Etiology Event Family Fatty Liver Fatty acid glycerol esters Fibrinogen Fibrosis Gene Expression General Population Genes Global Change Goals Hepatic Hepatocyte Histology Homeostasis Human In Vitro Individual Infiltration Inflammation Injection of therapeutic agent Injury to Liver Knock-out Knowledge Lead Length Leukocytes Life Lipid Peroxidation Lipids Liver Liver Cirrhosis Liver Failure Liver diseases MALAT1 gene Mediating Messenger RNA Metabolic Diseases Methods Modeling Molecular Monitor Mus Non-Insulin-Dependent Diabetes Mellitus Nuclear Obesity Oxidative Stress Pathology Pathway interactions Patients Phenotype Physiology Play Population Prevalence Protein Family Proteins RNA RNA Processing RNA Recognition Motif RNA Splicing RNA-Protein Interaction Reactive Oxygen Species Recombinants Regulation Regulator Genes Research Resolution Role Serine Spliced Genes Staging Stress Tail Tertiary Protein Structure Therapeutic Tissues Transcript Translations Untranslated RNA Up-Regulation Veins Viral Wild Type Mouse cell injury cytokine disease phenotype early onset effective therapy genetic regulatory protein in vivo insight liver function liver injury mRNA Precursor member mouse model mutant nonalcoholic steatohepatitis nucleocytoplasmic transport oxidative damage programs public health relevance research study response therapeutic development therapeutic target transcriptome transcriptome sequencing

项目摘要

DESCRIPTION (provided by applicant): Non-Alcoholic Steatohepatitis (NASH) is emerging as one of the most common liver disease in the American population. It is a metabolic disorder in which fat accumulation within the liver (steatosis) is associated with inflammation, hepatic injury and cirrhosis without significant consumption of alcohol. Despite affecting 2-5% of the American population, there are currently no effective therapeutic treatments for NASH. Current knowledge of this disease is limited because early stages (simple steatosis) are asymptomatic and difficult to detect. Furthermore, development of effective therapeutics against NASH pathology has been slow due to lack of a feasible and robust model system. We have discovered that hepatocyte-specific ablation of SRSF1 (SRSF1 HKO), a splicing regulatory protein, triggers severe and early onset of NASH phenotype. SRSF1 plays direct roles in both constitutive and alternative splicing and has recently been shown to also regulate translation and non-sense mediated decay of specific transcripts. Although the structural and functional roles of SRSF1 in splicing are extensively characterized, its role in tissue physiology is not well understood. The overall objective of this proposal is to determine the pathogenic mechanism(s) by which loss of SRSF1 results in NASH. Aim 1. We will first determine the underlying molecular irregularities promoting liver damage in our mouse model. Using an in vivo viral mediated SRSF1 HKO model, I will identify which of the activated mechanisms are primary versus secondary responses to loss of SRSF1 activity. We also have preliminary data which shows that MALAT1, a long non-coding RNA, has lost association to nuclear speckles in the SRSF1 HKO hepatocytes. Speckles are highly dynamic nuclear domains enriched with pre-mRNA splicing factors, RNA processing factors and RNA molecules including MALAT1. We will further investigate speckle composition in SRSF1 HKO hepatocytes and determine if loss of MALAT1 localization to speckles has direct implications in the development of NASH pathology. Aim 2. Secondly, we will construct the gene network regulated by SRSF1 by performing high-resolution RNA-Seq on hepatocytes isolated from wildtype and SRSF1 HKO mice. We will also determine direct mRNA targets of SRSF1 using iCLIP-Seq, a method used to identify protein-RNA interactions in living cells. Data from both of these approaches will allow for the construction ofa robust gene regulatory network. This network will provide insights into the molecular mechanisms resulting in the activation of cellular responses identified in Aim 1. Aim 3. Finally, we will determine which of SRSF1's functions, splicing or translation regulation, is crucial for maintaining normal hepatocyte function. This will be achieved using mutant constructs, which have either altered splicing and/or translation regulation activities. We will introduce these constructs to SRSF1 HKO livers in vivo and perform similar assays described in Aim 1. Results of these experiments will be compared to determine the contributions of SRSF1's splicing and translation regulatory functions in maintaining liver homeostasis.