Regulation of plasma LDL and HDL by microRNA-541-3p

microRNA-541-3p 对血浆 LDL 和 HDL 的调节

• 批准号: 10733641
• 负责人: M Mahmood Hussain
• 金额: $ 63.19万
• 依托单位: NYU LONG ISLAND SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10733641
• 关键词: 3' Untranslated Regions; Affect; Anti-Inflammatory Agents; Antiatherogenic; Antioxidants; Apolipoproteins; Apolipoproteins B; Applications Grants; Atherosclerosis; Binding Sites; Blood; Cardiovascular Diseases; Cause of Death; Cells; Cholesterol; Complex; Enhancers; Gene Expression Regulation; Genes; Genetic Transcription; Goals; Hepatic; Hepatocyte; High Density Lipoprotein Cholesterol; High Density Lipoproteins; Human; Libraries; Lipids; Lipoproteins; Low-Density Lipoproteins; Messenger RNA; MicroRNAs; Molecular; Mus; Mutate; Orthologous Gene; Output; Pathway interactions; Physiological; Plasma; Primary carcinoma of the liver cells; Production; Property; Proteins; RNA; Regulation; Reporting; Repression; Role; Site; Testing; Transcriptional Regulation; Untranslated RNA; Zinc Fingers; cardiovascular disorder risk; cardiovascular risk factor; combat; drug development; gene repression; hepatoma cell; in vivo; mRNA Transcript Degradation; mutant; new therapeutic target; novel; posttranscriptional; prevent; promoter; protein expression; transcription factor

High LDL and low HDL levels are risk factors for cardiovascular disease. Our long-term goal is to identify and explain mechanisms through which microRNAs (miRs) regulate plasma lipoprotein levels. Here, we report the identification of a novel miR-541-3p that decreases ApoB secretion and increases ApoA1 secretion in human hepatoma cells. Furthermore, we show that miR-541-3p regulates the zinc finger (Znf) transcription factors (TFs) zinc finger 101 (Znf101) and castor zinc finger protein 1 (Casz1), thereby modulating the expression of ApoB and ApoA1, respectively. On the basis of these novel findings, the following aims focus on elucidating how miR- 541-3p regulates the expression of these TFs, and how these TFs regulate ApoB and ApoA1 expression. Furthermore, we plan to establish the roles of these TFs in the control of plasma lipoproteins and atherosclerosis. Aim 1. Establish that miR-541-3p enhances post-transcriptional degradation of Znf101 and Casz1 mRNA by interacting with 3´-untranslated region (UTR) sequences. We propose to establish that RNA-RNA interactions between miR-541-3p and Znf101/Casz1 mRNAs are necessary for the regulation of mRNA levels and show that miR-541-3p interacts with target sequences in the 3´-UTR, thereby enhancing mRNA degradation. Aim 2. Explain the mechanism of regulation of ApoB and ApoA1 expression by Znf101 and Casz1 in human and mouse liver cells. We plan to elucidate how Znf101 and Casz1 regulate transcription of the APOB and APOA1 genes. We propose to show that Znf101 is an enhancer of APOB gene transcription, and Casz1 is a repressor of APOA1 gene transcription. Furthermore, we plan to clarify the roles of mouse orthologs of these TFs in the transcriptional regulation of mouse Apob and Apoa1 genes. These studies should reveal the molecular underpinnings of the regulation of ApoB and ApoA1 by miR-541-3p. Aim 3. Elucidate the roles of Zfp961 (mouse ortholog of human Znf101) and Casz1 in the regulation of plasma LDL, HDL, and atherosclerosis in mice. In these in vivo studies, we will investigate the physiological roles of ApoB and ApoA1 regulation by the TFs identified and characterized in the earlier aims. We will interrogate whether regulation of ApoB and ApoA1 by these TFs decreases LDL levels, increases HDL levels, and diminishes atherosclerosis. These studies should demonstrate that transcriptional regulation of ApoB and ApoA1 leads to an anti-atherogenic profile in mice. This proposal is based on our discovery of a novel miR-541-3p that regulates the expression of ApoB and ApoA1 in opposite directions through two newly identified TFs. After completion of the proposed studies, we expect to: (1) elucidate how miR-541-3p regulates various TFs; (2) explain the molecular mechanisms through which these TFs regulate APOB and APOA1 transcription; and (3) establish the physiological importance of the identified molecules and mechanisms in the regulation of plasma lipoproteins and atherosclerosis. These genes and mechanisms may serve as new therapeutic targets to prevent and treat cardiovascular disease.

高LDL和低HDL水平是心血管疾病的危险因素。我们的长期目标是确定和 解释microRNA（miRS）调节血浆脂蛋白水平的机制。在这里，我们报告 识别新型miR-541-3p，可减少APOB分泌并增加人类的ApoA1分泌 肝癌细胞。此外，我们表明miR-541-3p调节锌指（ZNF）转录因子（TFS） 锌指101（ZNF101）和蓖麻锌指蛋白1（CASZ1），从而调节APOB的表达 和apoa1。根据这些新发现，以下目的是阐明mir- 541-3p调节这些TF的表达，以及这些TF如何调节APOB和APOA1的表达。 此外，我们计划确定这些TF在控制血浆脂蛋白和动脉粥样硬化中的作用。 目标1。确定miR-541-3p增强了Znf101和Casz1 mRNA的转录后降解 通过与3´-非翻译区域（UTR）序列相互作用。我们建议确定RNA-RNA miR-541-3p与Znf101/casz1 mRNA之间的相互作用对于调节mRNA水平是必需的 并表明miR-541-3p与3´-UTR中的靶序列相互作用，从而增强mRNA降解。 AIM 2。解释Znf101和casz1中APOB和APOA1表达的调节机理 人和小鼠肝细胞。我们计划阐明Znf101和Casz1如何调节APOB的转录 和apoA1基因。我们建议表明Znf101是Apob基因转录的增强子，Casz1是 ApoA1基因转录的复制品。此外，我们计划阐明这些小鼠直系同源物的作用 小鼠APOB和APOA1基因的转录调控中的TF。这些研究应揭示分子 miR-541-3p对APOB和APOA1调控的基础。 目标3。阐明ZFP961（人Znf101的小鼠直系同源）和Casz1在调节中的作用 小鼠血浆LDL，HDL和动脉粥样硬化。在这些体内研究中，我们将研究生理 在早期目标中鉴定和表征的TFS调节APOB和APOA1的作用。我们将 询问这些TF对APOB和APOA1的调节是否会降低LDL水平，增加HDL水平， 并减少动脉粥样硬化。这些研究应证明APOB和APOB的转录调节 apoA1导致小鼠的抗动脉粥样硬化特征。 该提议基于我们发现的新型miR-541-3p，该miR-541-3p调节APOB和APOA1的表达 通过两个新鉴定的TF在相反的方向上。拟议的研究完成后，我们希望： （1）阐明miR-541-3p如何调节各种TF； （2）解释这些分子机制 TFS调节APOB和APOA1转录； （3）确定已确定的物理重要性 血浆脂蛋白和动脉粥样硬化调节中的分子和机制。这些基因和 机制可以作为预防和治疗心血管疾病的新治疗靶标。