Paracrine Action of BMP3 in Pulmonary Hypertension

BMP3 在肺动脉高压中的旁分泌作用

• 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
• 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

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 骨形成蛋白3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，其特征在于持续性毛细血管前病变， 肺动脉高压，导致右心衰竭和死亡。尽管在过去几十年里进行了大量的研究 PH仍然是一种具有高发病率和死亡率的不治之症。新的方向和疗法，以改善 我们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，我们目前对该途径几个成员的确切病理生理作用的理解 仍然有限。在本R 01中，我们提出了详细的功能和机制研究，以了解 PAH中的BMP 3。我们假设PASMC衍生的BMP 3抑制PAEC功能障碍和病理学改变。 肺血管重构基于强大的初步研究，我们建议测试这个中心 有三个具体目标的假设。 目的1将明确PAH中BMP 3的调节及其体外作用。我们将评估肺骨形态发生蛋白3的水平 肺组织和血清中的BMP 3水平和循环BMP 3水平 动物我们还将确定性别是否会影响人类和啮齿动物的肺BMP 3表达， 我们将研究PASMC衍生的BMP 3对PASMC和PAEC共培养物中PAEC功能的影响 从非PAH和PAH患者中分离。 目的2将表征细胞特异性BMP 3缺失和BMP 3上调的体内效应。我们将 提供了SMC和EC特异性BMP 3 KO的心肺表型的详细表征 小鼠我们还将在Sugen/缺氧模型中检测外源性重组BMP 3对PAH的影响。 对大鼠使用一种新的方法，我们已经证明是高度特异性和有效的蛋白质 在肺中过度表达（即化学修饰的mRNA（modRNA）），我们将评估BMP 3 modRNA逆转大鼠Sugen/缺氧模型中的PAH。 目的3：探讨BMP 3的分子机制。我们将进行一系列的体外和体内研究 确定BMP 3调节的机制并定义BMP 3作用机制。启动子 活性测定、免疫共沉淀实验、GST下拉测定、定量PCR测量 蛋白质印迹分析将确定BMP 3信号中涉及的途径。PASMC的共培养物和 来自BMP 3-KO小鼠和来自体内过表达BMP 3的大鼠的PAEC和肺将确定PAEC的活性。 与BMP 3信号有关的通路。