3'UTR Shortening in Pulmonary Vascular Disease

肺血管疾病中的 3UTR 缩短

• 批准号: 9921481
• 负责人: Harry Karmouty-Quintana
• 金额: $ 38.87万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-01 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9921481
• 关键词: 3' Untranslated Regions; Address; Affect; Apoptosis; Binding Sites; Biology; Blood Vessels; Blood flow; Cell Proliferation; Cellular Stress; Cessation of life; Chronic lung disease; Complex; Data; Deposition; Development; Diagnosis; Experimental Models; Extracellular Matrix; Extracellular Matrix Proteins; Fibrinogen; Functional disorder; Gene Expression; Genes; Heart failure; Human; Hypertrophy; In Vitro; Knockout Mice; Knowledge; Lead; Length; Lesion; Lung; Lung Transplantation; Lung diseases; Messenger RNA; Metabolism; MicroRNAs; Modeling; Molecular; Morbidity - disease rate; Mus; Pathogenesis; Pathologic Processes; Pathway Analysis; Pathway interactions; Patients; Physiological; Polyadenylation; Process; Proteins; Pulmonary Hypertension; Pulmonary artery structure; RNA; RNA-Binding Proteins; Regulation; Regulatory Element; Resistance; Right ventricular structure; Role; SM 22 muscle protein; Severity of illness; Side; Smooth Muscle; Smooth Muscle Myocytes; Structure of parenchyma of lung; System; Testing; Therapeutic Uses; Tissues; Transcript; Translations; Vascular Diseases; Vascular Smooth Muscle; Vascular remodeling; base; biobank; blood lead; cleavage factor; conditional knockout; effective therapy; hemodynamics; human tissue; in vivo; inhibitor/antagonist; knock-down; mortality; mouse model; nanoparticle delivery; novel; novel therapeutic intervention; novel therapeutics; overexpression; pressure; prevent; promoter; therapeutic evaluation; transcriptome sequencing

Project Summary Pulmonary hypertension (PH) is a disorder of the lung vasculature defined by increased mean pulmonary arterial pressure (mPAP) leading to right ventricle (RV) hypertrophy and dysfunction, right-sided heart failure and ultimately death. The pathologic process in PH is characterized by extensive vascular remodeling affecting pulmonary artery smooth muscle cells (PASMC) and the RV wall. Several process have been associated with the development of pulmonary hypertension including increased perivascular fibro-proliferative deposition altered cellular metabolism and increased cell proliferation and resistance to apoptosis. Recent advances in the field of RNA biology have shown that alternative polyadenylation (APA) results in shorter 3'-untranslated regions (3'UTR) of mRNAs that can avoid mRNA regulation resulting in overexpression of these transcripts. Recent studies have shown that 3'UTR is present following cellular stress. Depletion of a 25kDa subunit of the RNA binding protein cleavage factor I (CFIm25), has been shown to result in APA and marked 3'UTR shortening. Provocative preliminary data from my lab demonstrate depletion of CFIm25 and evidence of 3'UTR shortening from isolated pulmonary arteries from models of pulmonary hypertension. These observations are in line with evidence of 3'UTR shortening in isolated pulmonary artery smooth muscle cells. However, the effects of 3'UTR shortening in PH remain unknown. Pathway analysis from CFIm25 knock-down studies in PASMCs revealed 3'UTR shortening of genes associated with fibro-proliferative deposition and cellular proliferation. Taken together, our hypothesis is that 3'UTR shortening alters gene expression of many factors influencing the development of PH. Aim 1 will address how temporal changes in CFIm25 depletion lead to increased vascular remodeling through 3'UTR shortening. We will also evaluate expression levels of CFIm25 using patient-derived tissues and PASMCs to track how depletion of CFIm25 correlates with disease severity in PH. Using novel RNA- seq approaches in murine and human tissue we aim to identify changes in 3'UTR length in pathways contributing to the development of PH. In Aim 2, we will evaluate how CFIm25 depletion in vascular smooth muscle cells worsens the development of PH. Here we will also perform novel RNA-seq approaches to identify shortened 3'UTRs following depletion of CFIm25. Aim 3 will determine the levels of miRs-203 and miR-509-3p in patients with PH and in experimental models of PH. We will test the therapeutic potential of strategies aimed at elevating CFIm25 expression as a novel treatment for PH.

项目摘要 肺动脉高压（PH）是一种肺血管系统疾病， 压力（mPAP）导致右心室（RV）肥大和功能障碍、右侧心力衰竭和 最终死亡。PH的病理过程以广泛的血管重塑为特征， 肺动脉平滑肌细胞（PASMC）和RV壁。有几个过程与 肺动脉高压的发展，包括血管周围纤维增生性沉积增加 改变细胞代谢并增加细胞增殖和对凋亡的抗性。的最新进展 RNA生物学领域的研究表明，选择性多聚腺苷酸化（阿帕）导致较短的3 ′-非翻译区 在一些实施方案中，mRNA的非编码区（3 'UTR）可以避免mRNA调节，导致这些转录物的过表达。最近 研究表明，细胞应激后存在3’UTR。RNA的25 kDa亚基的缺失 结合蛋白切割因子I（CFIm 25）已显示导致阿帕和显著的3 'UTR缩短。 来自我实验室的挑衅性初步数据表明CFIm 25的耗尽和3 'UTR缩短的证据 从肺动脉高压模型中分离出的肺动脉。这些观察结果符合 分离的肺动脉平滑肌细胞中3 'UTR缩短的证据。然而，3 'UTR的作用 PH值的缩短仍然未知。来自PASMCs中CFIm 25敲低研究的途径分析显示， 与纤维增生性沉积和细胞增殖相关的基因的3 'UTR缩短。采取 总之，我们的假设是，3 'UTR缩短改变了许多影响基因表达的因素， 目的1将解决CFIm 25消耗的时间变化如何导致增加的血管生成。 通过3 'UTR缩短重塑。我们还将使用患者来源的免疫组织化学方法评估CFIm 25的表达水平。 组织和PASMC来追踪CFIm 25的缺失如何与PH疾病严重程度相关。使用新型RNA- 在鼠和人组织中的seq方法中，我们旨在鉴定3 'UTR长度的变化， 在目的2中，我们将评估血管平滑肌细胞中CFIm 25缺失如何影响PH的发展。 在这里，我们还将进行新的RNA-seq方法，以确定缩短的 CFIm耗尽后的3 'UTR 25。目的3将确定患者中miR-203和miR-509- 3 p的水平 与PH和PH的实验模型。我们将测试的治疗潜力的战略，旨在提高 CFIm 25表达作为PH的新治疗。