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REGULATION OF CHOLESTEROL HOMEOSTASIS BY NONCODING RNAS

非编码 RNA 调节胆固醇稳态

基本信息

批准号：
8274949
负责人：
DANIEL S ORY
金额：
$ 16.72万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-04-01 至 2014-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8274949
关键词：
Animal Model Atherosclerosis Cell Line Cells Chinese Hamster Ovary Cell Cholesterol Cholesterol Homeostasis Complement Computer Simulation Coupled Defect Disease Elements Endoplasmic Reticulum Equilibrium Event Exhibits Exons Fatty acid glycerol esters Feedback Functional RNA Gene Expression Gene Targeting Generations Genes Genetic Screening Genetic Translation Genomics Goals Hereditary Disease Human Human Genetics Inborn Errors of Metabolism Inborn Genetic Diseases Introns Knock-out Knockout Mice Laboratories Maintenance Mass Spectrum Analysis Measures Mediating Membrane Membrane Lipids Metabolic stress Metabolism MicroRNAs Modeling Molecular Molecular Target Nucleotides Oligonucleotides Open Reading Frames Orphan Pathway interactions Phenotype Physiological Predisposition Process Promoter Regions Proteomics RNA RNA Splicing Regulation Regulatory Pathway Role Small Nucleolar RNA Small RNA Sterols Time Translations Walking base cell growth regulation cell type cholesterol trafficking drug development embryonic stem cell feeding human DICER1 protein in vivo innovation lipoprotein cholesterol mRNA Precursor meetings mouse model mutant novel promoter public health relevance response trafficking uptake

项目摘要

DESCRIPTION (provided by applicant): Cellular cholesterol homeostasis is regulated at multiple cholesterol transfer steps and through a negative feedback loop that responds to elevations of membrane cholesterol in the endoplasmic reticulum (ER). Alterations in these sterol sensing and trafficking pathways in contribute to human inborn errors of metabolism and to acquired disease states. To elucidate mechanisms governing these critical cholesterol homeostatic pathways, we performed a functional genetic screen that led to isolation of cell lines with intracellular cholesterol trafficking defects that were enriched in mutants with disruption of long non-coding RNA (ncRNA) genes. These genes exhibit evolutionarily conserved exon/intron organization and core promoter regions, lack significant open reading-frames or nucleotide homology within exons, and contain short, highly conserved intronic regions that harbor small nucleolar RNA (snoRNA) species. This class of long ncRNAs appears principally to serve as host genes to facilitate expressing and processing of the orphan snoRNAs. We hypothesize that the processed snoRNAs modulate expression of genes involved in cholesterol homeostasis, possibly through control of splicing events or translation, and thus represent a previously unrecognized mode of regulation for cellular cholesterol homeostasis. The Specific Aims of this proposal are (1) To characterize ncRNA genes identified by our genetic screen that are critical for maintenance of cholesterol homeostasis, (2) To determine the role of the snoRNA host genes in regulation of intracellular cholesterol transfer and cellular cholesterol homeostasis, (3) To determine the molecular mechanism(s) through which the orphan snoRNA elements exert control over cholesterol regulatory pathways, and (4) To examine the physiological role of the orphan snoRNAs and gene pathways identified through the genetic screen by extending our cell-based studies to in vivo animal models. The proposed studies are innovative in that they explore a novel small RNA-dependent pathway not previously implicated in regulation of cellular cholesterol homeostasis. These studies are highly significant because elucidation of this RNA regulatory pathway has the potential to provide new molecular targets for manipulation of the cellular handling of cholesterol. This proposal is highly relevant to atherosclerosis, a common disease characterized by dysregulation of cholesterol homeostasis, as well as to rare, often fatal inborn errors of sterol metabolism. PUBLIC HEALTH RELEVANCE: Alterations in pathways that regulate cellular cholesterol balance contribute to human genetic diseases and to acquired disease states, such as atherosclerosis. Preliminary studies in our laboratory have identified a set of genes not previously implicated in these cholesterol regulatory pathways. The goal of this proposal is to study how these genes modulate the distribution and level of cholesterol in cells. The proposed studies may identify new targets for drug development for lowering cholesterol in disease states.

描述（由申请人提供）：细胞胆固醇稳态在多个胆固醇转移步骤中并通过响应内质网（ER）中膜胆固醇升高的负反馈回路进行调节。这些甾醇感应和运输途径的改变导致人类先天性代谢错误和后天疾病状态。为了阐明控制这些关键胆固醇稳态途径的机制，我们进行了功能性遗传筛选，分离出具有细胞内胆固醇运输缺陷的细胞系，这些缺陷在长非编码 RNA (ncRNA) 基因破坏的突变体中富集。这些基因表现出进化上保守的外显子/内含子组织和核心启动子区域，外显子内缺乏显着的开放阅读框或核苷酸同源性，并且含有短的、高度保守的内含子区域，其中包含小核仁RNA (snoRNA)物种。这类长 ncRNA 主要充当宿主基因，以促进孤儿 snoRNA 的表达和加工。我们假设加工后的 snoRNA 可能通过控制剪接事件或翻译来调节参与胆固醇稳态的基因表达，因此代表了一种以前未被识别的细胞胆固醇稳态调节模式。该提案的具体目标是 (1) 表征我们的遗传筛选鉴定出的对维持胆固醇稳态至关重要的 ncRNA 基因，(2) 确定 snoRNA 宿主基因在调节细胞内胆固醇转移和细胞胆固醇稳态中的作用，(3) 确定孤儿 snoRNA 元件控制胆固醇的分子机制 (4) 通过将我们基于细胞的研究扩展到体内动物模型，来检查通过遗传筛选确定的孤儿 snoRNA 和基因途径的生理作用。拟议的研究具有创新性，因为它们探索了一种新颖的小 RNA 依赖性途径，以前未涉及细胞胆固醇稳态的调节。这些研究非常重要，因为阐明这种 RNA 调节途径有可能为操纵胆固醇的细胞处理提供新的分子靶标。该提议与动脉粥样硬化高度相关，动脉粥样硬化是一种以胆固醇稳态失调为特征的常见疾病，以及罕见且通常致命的甾醇代谢先天性错误。公共卫生相关性：调节细胞胆固醇平衡途径的改变会导致人类遗传疾病和后天疾病状态，例如动脉粥样硬化。我们实验室的初步研究已经确定了一组以前未涉及这些胆固醇调节途径的基因。该提案的目标是研究这些基因如何调节细胞中胆固醇的分布和水平。拟议的研究可能会确定药物开发的新目标，以降低疾病状态下的胆固醇。