ROLE OF M-CSF IN THE PATHOGENESIS OF ATHEROSCLEROSIS

M-CSF 在动脉粥样硬化发病机制中的作用

• 批准号: 6625308
• 负责人: Tripathi Byasmuni Rajavashisth
• 金额: $ 30.6万
• 依托单位: CEDARS-SINAI MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-12-01 至 2004-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6625308
• 关键词: apolipoprotein E; atherosclerosis; atherosclerotic plaque; biological signal transduction; cell proliferation; colony stimulating factor; disease /disorder model; gene expression; genetic regulatory element; guanine nucleotide binding protein; immunocytochemistry; laboratory mouse; macrophage; mitogen activated protein kinase; molecular pathology; protein localization; protein structure function; radiotracer; site directed mutagenesis; stromelysin; tissue /cell culture; transcription factor; urokinase; vascular smooth muscle

Macrophage(MO)-colony stimulating factor (M-CSF) importantly contributes to the development of atherosclerotic lesions. We have found that the absence of M-CSF in atherosclerosis-prone apolipoprotein (apo) E or low-density lipoprotein receptor (LDLR)- deficient mice results in substantially reduced atherosclerosis despite augmented hypercholesterolemia. Our most recent studies provide compelling evidence in favor of a direct local effect of M-CSF within the vessel wall. These advances, together with the characterization of the M-CSF-mediated induction of urokinase plasminogen activator (uPA) and matrix metalloproteinases (MMPs) cascade have prompted more refined questions on the molecular mechanisms responsible for the full range of M-CSF actions in the diseased vessel wall. In this proposal, we seek to extend our research efforts to understand the role of M-CSF in the development and disruption of arterial lesions by testing following three hypotheses: 1) pleiotropic effects of M-CSF on intimal MO and SMC are modulated mainly through the activation of nuclear factors downstream to the Ras-mediated cell signaling pathways. One such factor is the transcription factor Ets-2 that promotes cell proliferation and survival, 2) increased M-CSF activity in atherosclerotic lesions contributes to the MO -mediated matrix remodeling by up regulating the expression of uPA and MT3-MMP genes. This effect of M-CSF may play a role in plaque disruption, and 3) M-CSF up regulates the MO-specific transcription of uPA and MT3-MMP genes by activating a common set of trans-acting factors (such as Ets family of transcription factors) that form ternary complexes with AP-l and bind to cis-acting DNA elements present in the 5' regulatory region of these genes. The specific aims are: 1) to investigate the effects of M-CSF on the growth of arterial lesion-associated cells in vivo using mice lacking M-CSF and/or apoE and to perform in vitro studies using cultured cells from M-CSF-deficient mice to determine the mechanism(s) underlying the M-CSF mediated proliferation and survival of MO and SMC, 2) to determine the effects of M-CSF on the expression of uPA and MT3-MMP in cultured MO and to examine the association of M-CSF regulated production of uPA and MT3-MMP to alterations in the character of atherosclerotic lesions, and 3) to identify the cis-acting elements in the uPA and MT3-MMP promoters that mediate the inductive effects of M-CSF on the expression of these genes and to examine the signaling events connecting M-CSF with the nuclear regulators of uPA and MT3-MMP. We believe our studies will provide new and important information regarding the role of M-CSF in the development and disruption of atherosclerotic and proliferative vascular lesions and this information may prove useful in design of novel therapeutic interventions for vascular diseases.

巨噬细胞（MO）-集落刺激因子（M-CSF）很重要 有助于动脉粥样硬化病变的发展。我们发现 易发生动脉粥样硬化的载脂蛋白 (apo) E 中缺乏 M-CSF 或 低密度脂蛋白受体（LDLR）缺陷的小鼠会导致 尽管高胆固醇血症增加，但动脉粥样硬化显着减少。 我们最近的研究提供了令人信服的证据，支持直接本地化 M-CSF 在血管壁内的作用。这些进步，连同 M-CSF 介导的尿激酶纤溶酶原诱导的表征 激活剂（uPA）和基质金属蛋白酶（MMP）级联促使更多 关于负责全方位的分子机制的精细问题 M-CSF 在患病血管壁中的作用。在本提案中，我们寻求扩展 我们的研究工作是为了了解 M-CSF 在发育和发育中的作用 通过测试以下三个假设来破坏动脉病变：1) M-CSF 对内膜 MO 和 SMC 的多效性作用主要通过调节 Ras 介导的细胞信号传导下游核因子的激活 途径。其中一种因子是转录因子 Ets-2，它可以促进细胞 增殖和存活，2) 动脉粥样硬化中 M-CSF 活性增加 病变通过上调 MO 介导的基质重塑 uPA 和 MT3-MMP 基因的表达。 M-CSF 的这种作用可能在 斑块破坏，3) M-CSF 上调 MO 特异性转录 uPA 和 MT3-MMP 基因通过激活一组常见的反式作用因子（例如 作为转录因子的 Ets 家族）与 AP-1 形成三元复合物 并与存在于 5' 调控区的顺式作用 DNA 元件结合 这些基因。具体目标是：1）研究M-CSF对 使用缺乏 M-CSF 的小鼠体内动脉病变相关细胞的生长 和/或 apoE 并使用来自的培养细胞进行体外研究 M-CSF 缺陷小鼠以确定 M-CSF 的潜在机制 介导的 MO 和 SMC 的增殖和存活，2) 确定效果 M-CSF 对培养 MO 中 uPA 和 MT3-MMP 表达的影响，并检查 M-CSF 调节 uPA 和 MT3-MMP 的产生与改变的关联 动脉粥样硬化病变的特征，3) 识别顺式作用 uPA 和 MT3-MMP 启动子中介导诱导效应的元件 M-CSF 对这些基因表达的影响并检查信号事件 将 M-CSF 与 uPA 和 MT3-MMP 的核调节因子连接起来。我们相信我们的 研究将提供有关 M-CSF 作用的新的重要信息 动脉粥样硬化和增殖性血管的发展和破坏 病变，这些信息可能有助于设计新的治疗方法 血管疾病的干预措施。