The mechanistic role of surfactant protein A in smooth muscle cell phenotype modulation and vascular remodeling

表面活性蛋白A在平滑肌细胞表型调节和血管重塑中的机制作用

• 批准号: 10516027
• 负责人: Skylar Dawn King
• 金额: $ 6.98万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-10 至 2024-09-09
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10516027
• 关键词: Angioplasty; Animals; Arterial Disorder; Arteries; Asthma; Atherosclerosis; Attenuated; Blood Vessels; Bypass; Carotid Arteries; Cell Differentiation process; Cell Nucleus; Cell Proliferation; Contractile Proteins; Data; Development; Diabetes Mellitus; Diabetic Angiopathies; Disease; Environment; Genes; Genetic Transcription; Goals; Histologic; Host Defense; Hyperplasia; In Vitro; Inflammatory Response; Injury; Knock-out; Knockout Mice; Laboratories; Lung; MYH11 gene; Malignant Neoplasms; Mechanics; Medial; Mediating; Mentors; Missouri; Modeling; Molecular; Mus; Operative Surgical Procedures; Pathogenesis; Phenotype; Phosphorylation; Physiological; Play; Process; Proteins; Pulmonary Surfactant-Associated Protein A; Rattus; Recombinants; Role; Signal Transduction; Smooth Muscle; Smooth Muscle Myocytes; Stents; Supplementation; Surface Tension; Systemic hypertension; Testing; Time; Training; Transforming Growth Factor beta; Transplantation; Universities; Vascular Diseases; Vascular Proliferation; Vascular Smooth Muscle; Vascular remodeling; alpha Actin; alveolar type II cell; extracellular; gain of function; genetic approach; human disease; hydrophilicity; in vivo; loss of function; mRNA Expression; mouse model; myocardin; neointima formation; neutralizing antibody; novel; promoter; restenosis; surfactant function; targeted treatment; therapy development; vascular injury

PROJECT SUMMARY The overall goal of this proposal is to find novel mechanisms whereby surfactant protein A (SPA) regulates vascular smooth muscle cell (SMC) phenotype modulation. SMC transition from a differentiated to dedifferentiated phenotype in addition to neointima formation/vascular remodeling has a critical role in human diseases such as the development of atherosclerosis, restenosis after angioplasty or bypass, diabetic vascular complications, arteriopathy transplants, asthma and cancer. Mechanisms that regulate SMC phenotype modulation and neointima formation are not well understood. The physiological function of SPA is its secretion by type II alveolar cells to maintain minimal surface tension in the lungs. However, preliminary data indicate a role for SPA as a SMC phenotype modulator. In vivo, SPA was expressed in the medial and neointimal SMCs following mechanical injury in rat and mouse carotid arteries. The wire-injury induced intimal hyperplasia was dramatically attenuated in SPA knockout mice. Furthermore, increased mRNA expression of SMC contractile genes and key regulators for contractile SMC phenotype, Myocardin and TGF-β1 was observed in SMCs isolated from SPA knockout mice. Additionally, SMCs from SPA knockout mice had increased Smad3 phosphorylation and the increase was blocked by the TGF-β1 neutralizing antibody. SPA is localized in the nucleus of SMCs suggesting it may have a role in SMC gene transcription. Indeed, SPA deficiency increased smooth muscle α-actin and smooth muscle 22-α promoter activity whereas recombinant SPA protein attenuated their activities. Hence, the central hypothesis is that SPA regulates SMC phenotype modulation and vascular remodeling through both extracellular (via modulating TGF-β1 signaling) and intracellular (Myocardin-related gene transcription) mechanisms. Using primary culture of SMC, in vivo mouse wire injury models combined with molecular, cellular and histological approaches, this proposal will 1) determine the molecular extracellular and intracellular mechanisms by which SPA regulates SMC phenotypic modulation; and 2) determine if SPA is essential for SMC phenotype modulation/vascular remodeling in vivo. Project completion will uncover novel mechanisms regulating SMC phenotypic modulation and provide understanding into whether SPA is a potential target for therapy against vascular damage associated with common vascular diseases such as diabetes, restenosis, atherosclerosis and cancer. The training plan laid out by the sponsor and the outstanding environment in the mentor’s laboratory and at the University of Missouri will safeguard the successful completion of the proposed studies.

项目摘要 该提案的总体目标是找到新的机制，表面活性剂蛋白A（SPA） 调节血管平滑肌细胞（SMC）表型调节。 SMC从一个 除了新内膜形成/血管重塑外，差异化为去分化的表型 在人类疾病中的关键作用，例如动脉粥样硬化的发展，再狭窄 血管成形术或旁路，糖尿病血管并发症，动脉疾病移植，哮喘和癌症。 调节SMC表型调制和新内膜形成的机制不好 理解齿。水疗中心的物理功能是由II型肺泡细胞分泌以维持 肺中的最小表面张力。但是，初步数据表明SPA作为SMC的作用 表型调制器。在体内，水疗中心在媒体和新媒介SMC中表达 大鼠和小鼠颈动脉的机械损伤。电线伤害引起的内膜增生为 在水疗敲除小鼠中动态减弱。此外，SMC的mRNA表达增加 收缩基因和收缩SMC表型，心肌蛋白和TGF-β1的关键调节剂是 在从水疗敲除小鼠中分离出的SMC中观察到。此外，来自水疗淘汰小鼠的SMC有 TGF-β1中和抗体阻止了SMAD3磷酸化的增加并增加。 水疗中心位于SMC的细胞核中，表明它可能在SMC基因转录中起作用。 实际上，水疗缺乏增加了平滑肌α-肌动蛋白和平滑肌22-α启动子活性 而重组水疗蛋白减弱了其活性。因此，中心假设是 SPA调节SMC表型调节和通过两个细胞外的血管重塑（通过 调节TGF-β1信号传导）和细胞内（肌心素相关基因转录）机制。 使用SMC的一级培养，体内小鼠电线损伤模型与分子，细胞和 组织学方法，该建议将确定细胞外和细胞内的分子 SPA调节SMC表型调制的机制； 2）确定水疗是否必不可少 用于体内的SMC表型调节/血管重塑。项目完成将发现小说 调节SMC表型调制的机制，并提供了理解水疗中水疗中的机制 治疗的潜在靶向与常见血管疾病相关的血管损伤的靶标 作为糖尿病，再狭窄，动脉粥样硬化和癌症。赞助商和 精神实验室和密苏里大学的杰出环境将维护 成功完成拟议的研究。