Mechanisms of PKG-induced PASMC Differentiation

PKG诱导PASMC分化的机制

• 批准号: 7212065
• 负责人: JESSE D ROBERTS
• 金额: $ 33.15万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-05-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7212065
• 关键词: Abbreviations; Adenoviruses; Apoptosis; Applications Grants; Arteries; Attenuated; Binding; Blood Vessels; Blood flow; Boxing; Cell Differentiation process; Cell Proliferation; Cells; Child; Childhood; Coiled-Coil Domain; Commercial Sources; Contractile Proteins; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Cyclic GMP; Cyclic GMP-Dependent Protein Kinases; Development; Diffuse; Distal; Endothelial Cells; Enzymes; Epitopes; Fingers; Gene Expression; Genes; Genetic Transcription; Green Fluorescent Proteins; Human; In Vitro; Infant; KT 5720; KT 5823; Laboratory Study; Lead; Length; Lung; Mediating; Membrane; Molecular; Morbidity - disease rate; Mus; N-terminal; Nature; Newborn Infant; Nitric Oxide; PKA inhibitor; Patients; Peripheral; Pharmaceutical Preparations; Phenotype; Phosphorylation; Phosphotransferases; Principal Investigator; Protein Isoforms; Proteins; Pulmonary Hypertension; Pulmonary artery structure; RNA Splicing; Regulation; Relaxation; Reporting; Research; Research Personnel; Role; Serum; Signal Pathway; Signal Transduction; Smooth Muscle Myocytes; Soluble Guanylate Cyclase; Specific qualifier value; Testing; Two-Hybrid System Techniques; Variant; Vascular Diseases; Yeasts; analog; base; genetic regulatory protein; human RBX1 protein; inhaled nitric oxide; inhibitor/antagonist; injured; lung injury; novel; novel therapeutics; prevent; programs; therapeutic target

DESCRIPTION (provided by applicant): Lung injury causes excessive pulmonary artery smooth muscle cell (PASMC) proliferation in the peripheral arteries of the lung and pulmonary hypertension and significant morbidity in infants and children. Laboratory studies indicate that inhaled nitric oxide (NO) attenuates pulmonary vascular disease in the developing lung. Although the protective mechanisms of NO are incompletely understood, NO regulates PASMC in part by stimulating soluble guanylate cyclase to synthesize cGMP, which stimulates cGMP-dependent protein kinase (PKG). PKG has been observed to induce PASMC differentiation by decreasing cell proliferation, increasing the expression of contractile proteins, and stimulating apoptosis. The BROAD, LONG-TERM OBJECTIVES of this grant proposal are to identify and characterize molecular mechanisms whereby PKG regulates PASMC differentiation. Recently, we identified two novel RING finger proteins, TRIM39R and axotrophin (AXOT), that interact with PKG in a yeast two-hybrid assay. Since PKG and RING finger proteins both regulate cell differentiation, they may share similar signaling pathways and RING finger proteins may mediate many of the cell regulatory activities of PKG. The central hypothesis of the proposal is that PKG regulates PASMC differentiation by interacting with RING finger proteins and regulatory proteins. Specific aim 1 examines how TRIM39R modulates PKG's regulation of PASMC proliferation, differentiation, and apoptosis. Specific aim 2 defines the molecular interaction between PKG and AXOT and defines the role of AXOT in modulating PKG's important non-vasodilatory actions in PASMC. Specific aim 3 utilizes a novel experimental approach to identify new RING finger and transcription regulating proteins phosphorylated by PKG in PASMC, based on their differential phosphorylation in cells. The studies proposed in this grant application will provide important new mechanistic information about how PKG modulates PASMC proliferation, differentiation, and apoptosis. They are likely to provide information that will lead to the development of novel therapeutic targets that can be used to prevent pulmonary vascular disease in the injured lung.

描述（由申请人提供）：肺损伤导致肺外周动脉中肺动脉平滑肌细胞（PASMC）过度增殖和肺动脉高压，以及婴儿和儿童的显着发病率。实验室研究表明，吸入一氧化氮 (NO) 可减轻发育中肺部的肺血管疾病。 尽管NO的保护机制尚不完全清楚，但NO部分通过刺激可溶性鸟苷酸环化酶合成cGMP来调节PASMC，cGMP刺激cGMP依赖性蛋白激酶(PKG)。据观察，PKG 通过减少细胞增殖、增加收缩蛋白的表达和刺激细胞凋亡来诱导 PASMC 分化。该拨款提案的广泛、长期目标是确定和表征 PKG 调节 PASMC 分化的分子机制。最近，我们鉴定了两种新型环指蛋白 TRIM39R 和轴肌营养不良蛋白 (AXOT)，它们在酵母双杂交测定中与 PKG 相互作用。由于 PKG 和 RING 指蛋白均调节细胞分化，因此它们可能具有相似的信号传导途径，并且 RING 指蛋白可能介导 PKG 的许多细胞调节活性。该提案的中心假设是 PKG 通过与 RING 指蛋白和调节蛋白相互作用来调节 PASMC 分化。具体目标 1 检查 TRIM39R 如何调节 PKG 对 PASMC 增殖、分化和凋亡的调节。具体目标 2 定义了 PKG 和 AXOT 之间的分子相互作用，并定义了 AXOT 在调节 PKG 在 PASMC 中的重要非血管舒张作用中的作用。 具体目标 3 利用一种新颖的实验方法，根据 PASMC 中 PKG 磷酸化的新 RING 指和转录调节蛋白，基于它们在细胞中的磷酸化差异。本拨款申请中提出的研究将提供有关 PKG 如何调节 PASMC 增殖、分化和凋亡的重要新机制信息。他们可能提供的信息将导致新的治疗靶点的开发，这些靶点可用于预防受损肺部的肺血管疾病。