Smooth muscle cell integration of differentiation signals

平滑肌细胞分化信号整合

• 批准号: 8644312
• 负责人: Lucy Liaw
• 金额: $ 38.34万
• 依托单位: MAINEHEALTH
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-03 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8644312
• 关键词: Address; Affect; Binding; Biochemical; Blood Vessels; Cardiovascular Diseases; Cell Communication; Cell Differentiation process; Cells; Characteristics; Clinical Trials; Complex; Core-Binding Factor; Data; Development; Disease; Disease model; Embryonic Development; Endoglin; Endothelial Cells; Family; Gene Expression; Gene Expression Regulation; Gene Targeting; Genes; Genetic; Goals; Growth; Human; Injury; Laboratories; Lesion; Ligands; Link; Mediating; Mediator of activation protein; Modeling; Molecular; Mus; Muscle; Mutate; Mutation; Paracrine Communication; Pathway interactions; Phenotype; Play; Regulation; Role; Signal Pathway; Signal Transduction; Smooth Muscle Actin Staining Method; Smooth Muscle Myocytes; TGF-beta type I receptor; Testing; Transcription Repressor/Corepressor; Transcriptional Regulation; Transgenic Mice; Transgenic Model; Vascular Diseases; Vascular Endothelial Cell; Vascular remodeling; cardiovascular disorder therapy; cytokine; gene function; in vitro Assay; in vivo; mouse model; notch protein; novel; receptor

DESCRIPTION (provided by applicant): The vascular smooth muscle cell (SMC) alters its molecular and phenotypic characteristics during pathological vascular remodeling. Both Notch and TGF¿ signaling pathways promote a differentiated, contractile phenotype characteristic of mature SMC. However, mechanisms by which these pathways interact with each other are only beginning to be understood. Mutations in either pathway are causal for human cardiovascular disease, and therapies targeting these pathways are currently in clinical trials. Our laboratory discovered a regulatory interaction between Notch and Smad in SMC, which links Notch and TGF¿/BMP/Smad signaling. Concomitant Notch and TGF¿ signaling leads to synergistic activation of phosphoSmad (pSmad) transcriptional activity and SMC marker expression. The goal of this project is to define mechanisms of integrative molecular signaling leading to SMC differentiation. This project utilizes molecular and biochemical signaling approaches in human primary vascular cells, and mouse transgenic models to study in vivo gene regulation and function. We propose the following hypotheses: 1) Jagged1 activation of Notch signaling in SMC transcriptionally represses the TGF¿ co-receptor, endoglin, via the canonical CBF-1 mediated pathway. 2) Coordinate Notch and TGF¿ signaling leads to synergistic activation of SMC contractile genes via interaction of CBF1- and pSmad- containing transcriptional complexes. 3) Activation of Notch in vivo in SMC prior to vascular injury will maintain the differentiated phenotype, leading to suppressed neointimal lesion formation and arteriogenesis. Specific Aim 1: Test the hypothesis that the TGF¿ co-receptor, endoglin, is transcriptionally regulated by Notch signaling in SMC. This aim will utilize molecular assays in vitro and transgenic mouse models in vivo to characterize Notch regulation of endoglin expression and TGF¿ signaling in SMC. Specific Aim 2: Characterize the interaction of CBF1 and Smad transcriptional complexes on SMC contractile genes. We hypothesize that CBF1 and pSmad transcriptional complexes interact to transcriptionally regulate contractile genes including smooth muscle ¿-actin (SM actin) and calponin1. Specific Aim 3: Determine how the regulation of endoglin and TGF¿ signaling by Notch affects pathological vascular remodeling. Established mouse transgenic strains will be evaluated in vascular disease models.

描述（由申请人提供）：血管平滑肌细胞（SMC）在病理性血管重塑过程中改变其分子和表型特征。 Notch 和 TGF¿ 信号通路均促进成熟 SMC 的分化、收缩表型特征。然而，这些途径相互作用的机制才刚刚开始被了解。任一途径的突变都会导致人类心血管疾病，针对这些途径的治疗目前正在进行临床试验。我们的实验室在 SMC 中发现了 Notch 和 Smad 之间的调节相互作用，该相互作用将 Notch 和 TGF¿/BMP/Smad 信号传导联系起来。伴随的 Notch 和 TGF¿ 信号传导导致 phosphaSmad (pSmad) 转录活性和 SMC 标记表达的协同激活。该项目的目标是定义导致 SMC 分化的综合分子信号传导机制。该项目利用人类原代血管细胞的分子和生化信号传导方法以及小鼠转基因模型来研究体内基因调控和功能。我们提出以下假设：1) Jagged1 激活 SMC 中的 Notch 信号传导，通过经典的 CBF-1 介导途径，转录抑制 TGF¿ 辅助受体内皮糖蛋白。 2) 协调Notch和TGF¿信号传导通过含有CBF1和pSmad的转录复合物的相互作用导致SMC收缩基因的协同激活。 3) 在血管损伤之前，SMC 体内 Notch 的激活将维持分化的表型，从而抑制新内膜病变的形成和动脉生成。具体目标 1：检验以下假设：TGF¿ 辅助受体内皮糖蛋白受 SMC 中的 Notch 信号传导转录调节。该目标将利用体外分子测定和体内转基因小鼠模型来表征 SMC 中内皮糖蛋白表达和 TGF¿ 信号传导的 Notch 调节。具体目标 2：表征 CBF1 和 Smad 转录复合物对 SMC 收缩基因的相互作用。我们假设 CBF1 和 pSmad 转录复合物相互作用以转录调节收缩基因，包括平滑肌 ¿-肌动蛋白 (SM 肌动蛋白) 和 calponin1。具体目标 3：确定 Notch 对内皮糖蛋白和 TGF¿ 信号传导的调节如何影响病理性血管重塑。已建立的小鼠转基因品系将在血管疾病模型中进行评估。