Specific Beta-tubulin Isotype Involvement in Foam Cell Formation and Progression of Atherosclerosis

项目总结/摘要 动脉粥样硬化性心血管疾病是全球发病率和死亡率的主要原因。电流 动脉粥样硬化的治疗集中于预防或血管修复， 导致动脉粥样硬化的进展。在细胞水平上，动脉粥样硬化的主要效应细胞是 当巨噬细胞经历与吞噬相关的形态学变化时形成的泡沫细胞， 脂质。这是由在细胞形状和功能中起重要作用的细胞骨架促进的。重要 细胞骨架的组分是由α-和β-二聚体聚合形成的管状聚合物， 微管蛋白叫做微管。微管的稳定性、动力学性质和行为取决于 β-微管蛋白同种型的组成，其中有许多。这些个体β-微管蛋白同种型的表达 在不同的细胞或组织类型中差异很大。我们假设β-微管蛋白同种型的一个特定亚类调节 泡沫细胞的形成和功能。为了了解β-微管蛋白同种型在泡沫细胞形成和功能中的作用， 我们提出了一种综合的方法，将体外和动物模型相结合，以了解 β-微管蛋白同种型在泡沫细胞形成和动脉粥样硬化发展中的生物学相关作用 斑块我们的目的是建立单个β-微管蛋白在泡沫细胞形成过程中的表达、分布和作用。 使用原代细胞（骨髓来源的巨噬细胞）体外形成，并确定β- 使用动脉粥样硬化小鼠模型进行体内微管蛋白依赖性泡沫细胞形成。β-微管蛋白同种型水平 在经历泡沫细胞形成的原代细胞和主动脉病变内测量细胞分布 使用β-微管蛋白同种型特异性抗体。将在细胞和主动脉组织中定量β-微管蛋白基因表达 使用qRT-PCR。此外，改变原代细胞中个别β-微管蛋白同种型或β-微管蛋白 结合药物在小鼠中，对泡沫细胞的影响将被评估。拟议的研究将确定特定的β-微管蛋白 参与泡沫细胞形成的同种型。我们将获得的知识可以用来设计特定的药物， 通过靶向特定β-微管蛋白同种型抑制泡沫细胞形成和随后的斑块形成。

Project Summary/Abstract Atherosclerotic cardiovascular disease is the leading cause of morbidity and mortality worldwide. Current treatments for atherosclerosis focus on prevention or vessel repair with very few targeting the cellular events leading to the progression of atherosclerosis. At the cellular level, the main effector cell of atherosclerosis is the foam cell which is formed when macrophages undergo a morphological change associated with engulfment of lipids. This is facilitated by the cytoskeleton which plays important roles in cell shape and function. Important components of the cytoskeleton are the tubular polymers formed by the polymerization of dimers of α- and β- tubulin called microtubules. The stability, dynamic properties, and behavior of a microtubule depend on the composition of β-tubulin isotypes, of which there are many. The expression of these individual β-tubulin isotypes can differ greatly across cell or tissue type. We hypothesize that a specific subset of β-tubulin isotypes regulates foam cell formation and function. To understand the role of β-tubulin isotypes in foam cell formation and function, we propose a comprehensive approach integrating in vitro and animal models to gain an understanding of the biologically relevant roles β-tubulin isotypes play in foam cell formation and the development of atherosclerotic plaques. We aim to establish the expression, distribution, and role of individual β-tubulins during foam cell formation in vitro using primary cells (bone-marrow derived macrophage) and to determine the mechanism of β- tubulin-dependent foam cell formation in vivo using mouse models of atherosclerosis. β-tubulin isotype levels and cellular distribution will be measured in primary cells undergoing foam cell formation and within aortic lesions using β-tubulin isotype-specific antibodies. β-tubulin gene expression will be quantified in cells and aortic tissue using qRT-PCR. In addition, the effect that altering individual β-tubulin isotypes in primary cells or, with β-tubulin binding drugs in mice, has on foam cells will be assessed. The proposed study will define the specific β-tubulin isotypes involved in foam cell formation. Knowledge that we will gain could be used to design specific drugs that inhibit foam cell formation, and subsequent plaque formation, by targeting specific β-tubulin isotypes.