Paracrine Action of BMP3 in Pulmonary Hypertension

BMP3 在肺动脉高压中的旁分泌作用

基本信息

批准号：
10683927
负责人：
Yassine Sassi
金额：
$ 44.03万
依托单位：
VIRGINIA POLYTECHNIC INST AND ST UNIV
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-15 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10683927
关键词：
Affect Age Animal Diseases Animal Model Animals Apoptosis BMP3 gene Biological Assay Blood capillaries Bone Morphogenetic Proteins Cardiopulmonary Cell Communication Cell Proliferation Cell Separation Cell physiology Cell secretion Cells Cessation of life Chemicals Clinical Co-Immunoprecipitations Coculture Techniques Disease Down-Regulation Endothelial Cells Endothelin-1 Equilibrium Exposure to Functional disorder Future Goals Heritability Heterozygote Human Hypertension Hypoxia In Vitro Injury Knockout Mice Ligands LoxP-flanked allele Lung MADH2 gene MADH4 gene Measurement Messenger RNA Modeling Molecular Morbidity - disease rate Mus Mutation Pathogenesis Pathologic Pathway interactions Patients Pharmaceutical Preparations Phase Phenotype Phosphorylation Pilot Projects Process Production Protein Overexpression Pulmonary Hypertension Pulmonary artery structure Rattus Recombinants Regulation Research Rodent Role Series Serum Signal Pathway Signal Transduction Smooth Muscle Myocytes Testing Up-Regulation Vascular Diseases Vascular remodeling Western Blotting bone morphogenetic protein receptors experimental study improved in vivo intercellular communication loss of function mutation member migration mortality new therapeutic target novel strategies novel therapeutic intervention overexpression paracrine promoter pulmonary arterial hypertension pulmonary vascular remodeling pulmonary vasoconstriction right ventricular failure sex treatment strategy

项目摘要

PROJECT ABSTRACT/SUMMARY Paracrine Action of BMP3 in Pulmonary Hypertension Pulmonary arterial hypertension (PAH) is a severe vascular disease characterized by persistent precapillary pulmonary hypertension, leading to right heart failure and death. Despite intense research in the last decades PH remains an incurable disease with a high morbidity and mortality. New directions and therapies to improve understanding and treatment of PAH are desperately needed. Although evidence demonstrates the importance of the Bone Morphogenetic Protein (BMP) signaling pathway in PAH, our current understanding of the exact pathophysiological role of several members of this pathway remains limited. In this R01 we propose detailed functional and mechanistic studies to understand the role of BMP3 in PAH. We hypothesize that PASMC-derived BMP3 inhibits PAEC dysfunction and pathological pulmonary vascular remodeling. Based on robust preliminary studies, we propose testing this central hypothesis with 3 specific aims. Aim 1 will define BMP3 regulation in PAH and its effects in vitro. We will assess the levels of pulmonary BMP3 levels and circulating BMP3 levels in lung and serum of patients with clinical PAH and of PAH-diseased animals. We will also determine whether sex affects pulmonary BMP3 expression in humans and rodents, and we will investigate the effect of PASMC-derived BMP3 on PAEC function in cocultures of PASMCs and PAECs isolated from non-PAH and PAH patients. Aim 2 will characterize the in vivo effects of cell-specific BMP3 deletion and of BMP3 upregulation. We will provide a detailed characterization of the cardiopulmonary phenotypes of SMC- and EC-specific BMP3 KO mice. We will also examine the effects of exogenous recombinant BMP3 on PAH in the Sugen/Hypoxia model in rats. Using a novel approach, which we have demonstrated to be highly specific and efficient for protein overexpression in the lung (i.e. chemically modified mRNAs (modRNAs)), we will assess whether BMP3 modRNA reverses PAH in the Sugen/Hypoxia model in rats. Aim 3 will dissect the molecular mechanisms of BMP3. We will perform a series of in vitro and in vivo studies to determine the mechanism(s) of BMP3 regulation and to define BMP3 mechanism(s) of action. Promoter activity assays, co-immunoprecipitation experiments, GST pull down assays, quantitative PCR measurements and western blot analyses will determine the pathways implicated in BMP3 signals. Cocultures of PASMCs and PAECs and lungs from BMP3-KO mice and from rats overexpressing BMP3 in vivo will determine the pathways implicated in BMP3 signals.

项目摘要/总结 BMP3 在肺动脉高压中的旁分泌作用肺动脉高压（PAH）是一种严重的血管疾病，其特征是持续性毛细血管前病变肺动脉高压，导致右心衰竭和死亡。尽管过去几十年进行了大量研究 PH 仍然是一种发病率和死亡率较高的不治之症。改进的新方向和疗法迫切需要了解和治疗 PAH。尽管有证据表明骨形态发生蛋白 (BMP) 信号通路的重要性在 PAH 中，我们目前对该途径的几个成员的确切病理生理学作用的理解仍然有限。在本 R01 中，我们提出了详细的功能和机制研究，以了解 PAH 中的 BMP3。我们假设 PASMC 衍生的 BMP3 抑制 PAEC 功能障碍和病理学肺血管重塑。基于强有力的初步研究，我们建议测试这个中心具有 3 个具体目标的假设。目标 1 将定义 BMP3 在 PAH 中的调节及其体外作用。我们将评估肺部 BMP3 的水平临床 PAH 患者和 PAH 疾病患者肺和血清中的水平和循环 BMP3 水平动物。我们还将确定性别是否影响人类和啮齿动物的肺 BMP3 表达，以及我们将研究 PASMC 衍生的 BMP3 对 PASMC 和 PAEC 共培养中 PAEC 功能的影响从非 PAH 和 PAH 患者中分离出来。目标 2 将描述细胞特异性 BMP3 缺失和 BMP3 上调的体内效应。我们将提供 SMC 和 EC 特异性 BMP3 KO 心肺表型的详细特征老鼠。我们还将在 Sugen/缺氧模型中研究外源重组 BMP3 对 PAH 的影响在老鼠身上。使用一种新方法，我们已证明该方法对蛋白质具有高度特异性和高效性如果肺中过度表达（即化学修饰的 mRNA (modRNA)），我们将评估 BMP3 是否 modRNA 在大鼠 Sugen/缺氧模型中逆转 PAH。目标 3 将剖析 BMP3 的分子机制。我们将进行一系列体外和体内研究确定 BMP3 调节机制并定义 BMP3 作用机制。发起人活性测定、免疫共沉淀实验、GST Pull Down 测定、定量 PCR 测量蛋白质印迹分析将确定 BMP3 信号所涉及的途径。 PASMC 的共培养和来自 BMP3-KO 小鼠和体内过表达 BMP3 的大鼠的 PAEC 和肺将决定与 BMP3 信号有关的通路。