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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：
8444326
负责人：
DANIEL S ORY
金额：
$ 35.81万
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 2015-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8444326
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.

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通过对内质网（ER）中膜胆固醇升高做出响应的负反馈回路进行调节。这些固醇传感和运输途径的改变有助于人类先天性代谢缺陷和获得性疾病状态。为了阐明这些关键的胆固醇稳态途径的机制，我们进行了功能性遗传筛选，导致细胞内胆固醇运输缺陷的细胞系的分离，这些细胞系富含长非编码RNA（ncRNA）基因破坏的突变体。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，缺乏显着的开放阅读框或核苷酸同源性的外显子内，并含有短，高度保守的内含子区域，港口小核仁RNA（snoRNA）的物种。这类长ncRNA似乎主要用作宿主基因以促进孤儿snoRNA的表达和加工。我们假设，加工snoRNA调节参与胆固醇稳态的基因的表达，可能通过控制剪接事件或翻译，因此代表了一种以前未被认识到的细胞胆固醇稳态调节模式。该提议的具体目的是（1）表征通过我们的遗传筛选鉴定的对于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元件通过其对胆固醇调节途径施加控制的分子机制，以及（4）通过将我们的基于细胞的研究扩展到体内动物模型来检查孤儿snoRNA的生理作用和通过遗传筛选鉴定的基因通路。拟议的研究是创新的，因为他们探索了一种新的小RNA依赖性途径，以前没有涉及细胞胆固醇稳态的调节。这些研究是非常重要的，因为这种RNA调节途径的阐明有可能提供新的分子靶点，用于操纵细胞处理胆固醇。这一建议与动脉粥样硬化（一种以胆固醇稳态失调为特征的常见疾病）以及罕见的、往往致命的甾醇代谢先天性缺陷高度相关。