Expression of PKG in Vascular Smooth Muscle Cells

PKG在血管平滑肌细胞中的表达

• 批准号: 8108392
• 负责人: Thomas M. Lincoln
• 金额: $ 37.13万
• 依托单位: UNIVERSITY OF SOUTH ALABAMA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-01-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8108392
• 关键词: Accounting; Address; Animal Model; Atherosclerosis; Blood Vessels; Blood flow; Cardiovascular Diseases; Carotid Arteries; Cell Culture Techniques; Cells; Cessation of life; Contractile Proteins; Contracts; Cultured Cells; Cyclic GMP; Cyclic GMP-Dependent Protein Kinases; Disease; Down-Regulation; Enzymes; Excision; Extracellular Matrix; Extracellular Matrix Proteins; Failure; Funding; Gene Expression; Genetic; Growth Factor; Immunoprecipitation; In Vitro; Inflammation; Inflammation Mediators; Inflammatory; Injury; Interleukin-1; Ischemia; Knockout Mice; Knowledge; Laboratories; Lead; Learning; Lesion; Ligase; Light; Lysine; Mediator of activation protein; Messenger RNA; Modeling; Nitric Oxide; Nitric Oxide Signaling Pathway; Nitric Oxide Synthase; Organ; Pathway interactions; Phenotype; Platelet Activation; Platelet aggregation; Platelet-Derived Growth Factor; Procedures; Proliferating; Protein Isoforms; Protein-Serine-Threonine Kinases; Proteins; Research; Role; Signal Pathway; Signal Transduction; Site-Directed Mutagenesis; Smooth Muscle; Smooth Muscle Myocytes; Soluble Guanylate Cyclase; Stimulus; Stroke; TNF gene; Testing; Tissues; Transfection; Vascular Diseases; Vascular Smooth Muscle; Vascular remodeling; Work; Wound Healing; cytokine; genetic pedigree; human NOS2A protein; in vivo; inhibitor/antagonist; interest; nitric oxide synthase II; premature; prevent; protein expression; response; response to injury; restenosis; restoration; ubiquitin-protein ligase; vascular smooth muscle cell proliferation; Accounting; Address; Animal Model; Atherosclerosis; Blood Vessels; Blood flow; Cardiovascular Diseases; Carotid Arteries; Cell Culture Techniques; Cells; Cessation of life; Contractile Proteins; Contracts; Cultured Cells; Cyclic GMP; Cyclic GMP-Dependent Protein Kinases; Disease; Down-Regulation; Enzymes; Excision; Extracellular Matrix; Extracellular Matrix Proteins; Failure; Funding; Gene Expression; Genetic; Growth Factor; Immunoprecipitation; In Vitro; Inflammation; Inflammation Mediators; Inflammatory; Injury; Interleukin-1; Ischemia; Knockout Mice; Knowledge; Laboratories; Lead; Learning; Lesion; Ligase; Light; Lysine; Mediator of activation protein; Messenger RNA; Modeling; Nitric Oxide; Nitric Oxide Signaling Pathway; Nitric Oxide Synthase; Organ; Pathway interactions; Phenotype; Platelet Activation; Platelet aggregation; Platelet-Derived Growth Factor; Procedures; Proliferating; Protein Isoforms; Protein-Serine-Threonine Kinases; Proteins; Research; Role; Signal Pathway; Signal Transduction; Site-Directed Mutagenesis; Smooth Muscle; Smooth Muscle Myocytes; Soluble Guanylate Cyclase; Stimulus; Stroke; TNF gene; Testing; Tissues; Transfection; Vascular Diseases; Vascular Smooth Muscle; Vascular remodeling; Work; Wound Healing; cytokine; genetic pedigree; human NOS2A protein; in vivo; inhibitor/antagonist; interest; nitric oxide synthase II; premature; prevent; protein expression; response; response to injury; restenosis; restoration; ubiquitin-protein ligase; vascular smooth muscle cell proliferation

DESCRIPTION (provided by applicant): Atherosclerosis and other vascular diseases account for more deaths in the U.S. than any other cause. It is now well-understood that atherosclerosis is an inflammatory disorder whereby in response to injury and insult to the vasculature, growth factors and cytokines stimulate normal vascular smooth muscle cells (VSMC) to proliferate and secrete excessive extracellular matrix material. The resultant lesions cause disruptions in normal blood flow, platelet aggregation and activation, and eventually end-organ damage due to circulatory failure. Much research in the past two decades has identified signaling pathways in VSMC that lead to these proliferative and secretory changes. One pathway that normally blocks VSMC proliferation and secretory activity and promotes normal gene expression is the nitric oxide (NO)/cGMP pathway. Cyclic GMP activates a serine/threonine protein kinase, PKG-I, in vascular smooth muscle that leads to increases in normal contractile protein gene expression and suppresses proliferative and secretory activity. PKG-I thus is an important homeostatic control point whose activity is important for preventing excessive VSMC phenotypic modulation to the fibroproliferative state. Over the past few years, our laboratory has been interested in the mechanisms that control the expression of PKG-I in VSMC. Briefly, PKG-I expression is suppressed by inflammatory cytokines and growth factors that increase the fibroproliferative phenotype; restoration of PKG-I expression by transfection/adenoviral transduction restores the contractile phenotype of the VSMC. Hence, how inflammation controls PKG-I expression is critically important to understand in order to fully understand how these vascular disorders come about. During the previous funding period, we identified a pathway by which inflammatory cytokines regulate PKG-I expression. By inducing the expression of type II NO synthase (iNOS) in VSMC, cytokines/growth factors cause a persistent elevation in cGMP which directly causes down-regulation in the levels of PKG-I. This, in turn, is caused by cGMP-induced autophosphorylation of the PKG-Ia isoform and its ubiquitinylation. In this proposal, we will investigate the mechanisms by which PKG-Ia is ubiquitinylated in cultured VSMC, and determine whether this mechanism exists in vivo using wild- type and iNOS null mice to define the role of iNOS. The results of this proposed three-year study will shed new light on the mechanisms by which inflammation bring about vascular disorders. PUBLIC HEALTH RELEVANCE: Cardiovascular diseases such as atherosclerosis and stroke account for more premature deaths than any other cause. This project will investigate the genesis of atherosclerosis by studying the role of a signaling pathway (nitric oxide-cyclic GMP) in vascular smooth muscle cells in a genetic animal model. Specifically, a key component of this pathway (PKG) will be studied to understand how inflammation leads to these vascular disorders.

描述（由申请人提供）：在美国，动脉粥样硬化和其他血管疾病的死亡人数比任何其他原因都要多。现在，人们对动脉粥样硬化是一种炎症性疾病的理解，响应损伤并侮辱脉管系统，生长因子和细胞因子刺激正常的血管平滑肌细胞（VSMC），以增殖并分泌过多的细胞外基质材料。最终的病变会导致正常血流，血小板聚集和激活以及由于循环衰竭而导致的最终器官损伤。在过去的二十年中，许多研究都确定了VSMC中的信号通路，从而导致了这些增殖和分泌变化。通常阻止VSMC增殖和分泌活性并促进正常基因表达的一种途径是一氧化氮（NO）/CGMP途径。环状GMP在血管平滑肌中激活丝氨酸/苏氨酸蛋白激酶PKG-I，从而导致正常收缩蛋白基因表达的增加并抑制增殖和分泌活性。因此，PKG-I是一个重要的稳态控制点，其活性对于防止过度的VSMC表型调节对纤维增生状态很重要。在过去的几年中，我们的实验室一直对控制PKG-I在VSMC中的表达的机制感兴趣。简而言之，炎症细胞因子和增加纤维增生性表型的生长因子抑制了PKG-I的表达。通过转染/腺病毒转导恢复PKG-I表达，恢复了VSMC的收缩表型。因此，炎症如何控制PKG-I表达至关重要，以便完全理解这些血管疾病是如何产生的。在上一个资金期间，我们确定了炎症细胞因子调节PKG-I表达的途径。通过在VSMC中诱导II型NO合酶（INOS）的表达，细胞因子/生长因子导致CGMP持续升高，这直接导致PKG-I水平下调。反过来，这是由CGMP诱导的PKG-IA同工型及其泛素化的自磷酸化引起的。在此提案中，我们将研究PKG-IA在培养的VSMC中被泛素化的机制，并确定使用野生型和Inos null小鼠在体内是否存在这种机制来定义INOS的作用。这项提出的三年研究的结果将为炎症引起血管疾病的机制提供新的启示。 公共卫生相关性：心血管硬化和中风等心血管疾病比任何其他原因都要过早死亡。该项目将通过研究遗传动物模型中信号传导途径（一氧化氮循环GMP）在血管平滑肌细胞中的信号传导途径（一氧化氮循环GMP）的作用来研究动脉粥样硬化的发生。具体而言，将研究该途径（PKG）的关键组成部分，以了解炎症如何导致这些血管疾病。