Determining roles of the pro-apoptotic UPR gene CHAC1 in atherosclerosis.

确定促凋亡 UPR 基因 CHAC1 在动脉粥样硬化中的作用。

• 批准号: 8532027
• 负责人: Imran Mungrue
• 金额: $ 23.54万
• 依托单位: LSU HEALTH SCIENCES CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8532027
• 关键词: Ablation; Americas; Apoptosis; Apoptotic; Arterial Fatty Streak; Atherosclerosis; Binding; Biological Assay; Biology; Blood Vessels; Cell physiology; Cells; Clear Cell; Cultured Cells; DNA; Development; Diet; Disease; Disease model; Generations; Genes; Genetic; Grant; Heart Arrest; Knock-out; Knockout Mice; Lesion; Mediating; Messenger RNA; Methods; Molecular; Morbidity - disease rate; Mus; Pathway interactions; Peptides; Plasmids; Precipitation; Process; Protein Region; Proteins; Role; Signal Pathway; Signal Transduction; Site-Directed Mutagenesis; Small Interfering RNA; Smooth Muscle Myocytes; Stroke; Systems Biology; TNFRSF6B gene; TNFSF6 gene; Vascular Diseases; base; caspase 12; in vivo; macrophage; mortality; novel; promoter; receptor; research study; response; screening

Atherosclerosis is the major cause of morbidity and mortality in America. This disease process is defined by the formation of vascular lesions that contain activated macrophages (MO) and dysfunctional endothelial (EC) and smooth muscle cells (SMC). The unfolded protein response (UPR) has been implicated as a cellular process that is induced in atherosclerotic lesions, and in cultured cells (MO, EC and SMC) treated with known pro-atherogenic inducers. The primary aim of this grant will be to explore the function of a novel gene, CHAC1. This protein is specifically induced by UPR activation and is downstream ofthe ATF4-ATF3- CHOP cascade, an important pro-apoptotic pathway ofthe UPR. CHAC1 is a soluble cytosolic peptide, which induces apoptosis when over-expressed, and siRNA knockdown confirms a pro-apoptotic role for this gene. CHAC1 activates TNFRSF6B, a FASL decoy receptor, which functions to antagonize FASL-FAS induced apoptosis signaling. This pathway is of importance because FASL expressing cells can induce apoptosis of MO, EC or SMC, and this my exacerbate the progression of vascular disease. The proposal herein will define the function of CHAC1 at the molecular level, and characterize its contribution to apoptosis and the atherosclerotic disease process in MO, EC and SMC. The contribution of CHAC1 in CHOP and UPR induced apoptosis will be defined using established over-expression plasmids and siRNAs, generation of knockout cells, and assaying different components of putative signaling cascades. Direct binding partners of CHAC1 will also be explored using the TAP method, to define the molecular'^function ofthis gene. Candidates from TAP screening will be validated using co-immuno-precipitation. Based on candidates identified, pijtative functions ofthe chaC domain will be assayed, and targeted deletion and site directed mutagenesis used to define important regions of the protein. Finally, the use of mice that have the CHAC1 gene knocked out will be generated to explore the effects of CHAC1 genetic ablation in the development of atherosclerosis in a mouse disease model.

动脉粥样硬化是美国发病率和死亡率的主要原因。这种疾病的过程被定义为