Novel molecular mechanisms of vascular smooth muscle cell-mediated large and small artery calcification

项目摘要/摘要 本提案描述了一项为期5年的心血管学术生涯发展培训计划。 研究。利诺·卡德纳斯博士将在一个超过100人的环境中从多名调查人员的专业知识中获益 并将使用MGH/HMS的一系列生物医学核心设施，以帮助成功 完成申请书中概述的研究计划。利诺·卡德纳斯博士的研究项目将重点放在 与急性冠脉病变密切相关的血管钙化分子机制研究 综合征、动脉粥样硬化和外周动脉疾病。越来越多的证据指向自噬，一种 进化上保守的过程，如在动脉粥样硬化早期起保护作用。然而，自噬可以 随着动脉粥样硬化的进展而变得失调。在初步研究中，利诺·卡德纳斯博士观察到 自噬途径的药物激活减少了MGP-/-小鼠(小鼠)的血管钙化 自发性血管钙化模型)并提高存活率。候选人提议定义 破坏自噬加剧血管钙化的确切分子机制 以下两个目标：在目标1中，候选人的第一个目标是确定血管平滑肌的作用 细胞(VSMC)特异性自噬失调对血管钙化发展的影响 自噬的调节可能会抑制血管钙化。利诺·卡德纳斯博士将扩展他的初步发现 通过结合RNA-SEQ、CHIP-SEQ和ATAC-SEQ技术来定义染色质可塑性对 血管钙化过程中的自噬途径。利诺·卡德纳斯博士将研究一系列 体外和体外自噬调节剂对VSMC表型和钙化的影响。在目标2中，候选人将 确定自噬途径的药物激活或遗传激活是否抑制血管钙化 在体内使用两种不同的小鼠血管钙化模型。增进对新分子的认识 血管钙化的机制可能具有重要的临床意义并提供新的 心血管疾病的治疗目标。候选人的目标是立即完成以下工作 和长期的职业目标：(1)更广泛地了解导致 血管钙化。(2)学习利用现有的人类基因数据库进行基因组-表型分析 鉴定与血管表型相关的自噬启动基因的多态。此外，利诺博士 卡德纳斯将学习分子生物学和多只小鼠条件性基因缺失的先进技术 模特们。(3)在导师和咨询委员会的指导下，培养必要的技能 指导实验室，促进富有成效的研究合作和拨款撰写(4)以成功申请 在他开始获奖后的3年内申请R01资金。

PROJECT SUMMARY/ABSTRACT This proposal describes a 5-year training program for the development of an academic career in cardiovascular research. Dr. Lino Cardenas will gain from the expertise of multiple investigators in an environment of over 100 faculty and will have access to a wide range of biomedical core facilities at MGH/HMS to aid in the successful completion of the research program outlined in his application. Dr. Lino Cardenas's research program focuses on the molecular mechanisms of vascular calcification, highly relevant to the pathogenesis of acute coronary syndromes, atherosclerosis, and peripheral arterial disease. Growing evidence points towards autophagy, an evolutionarily conserved process, as being protective during early atherosclerosis. However, autophagy can become dysregulated with advanced atherosclerosis. In preliminary studies, Dr. Lino Cardenas has observed that pharmacologic activation of the autophagy pathway reduces vascular calcification in Mgp-/- mice (mouse model of spontaneous vascular calcification) and improves survival. The candidate proposes to define the precise molecular mechanisms by which disruption of autophagy exacerbates vascular calcification with the following two aims: In Aim 1, the candidate's first objective is to determine the role of vascular smooth muscle cell (VSMC)-specific autophagy dysregulation on the development of vascular calcification and whether modulation of autophagy may inhibit vascular calcification. Dr. Lino Cardenas will extend his preliminary findings by combining RNA-seq, ChIP-seq and ATAC-seq technologies to define the effects of chromatin plasticity on the autophagy pathway during vascular calcification. Dr. Lino Cardenas will study the therapeutic effect of a range of autophagy modulators on VSMC phenotype and calcification in vitro and ex vivo. In Aim 2, the candidate will determine whether pharmacologic or genetic activation of the autophagy pathway inhibits vascular calcification in vivo using two different murine models of vascular calcification. Enhancing knowledge of novel molecular mechanisms responsible for vascular calcification may hold important clinical implications and provide new targets for the treatment of cardiovascular disease. The candidate aims to accomplish the following immediate and long-term career goals: (1) To develop a broader understanding of the molecular mechanisms resulting in vascular calcification. (2) To learn genomice-phenotype analyses using existing human genetic databases to identify polymorphisms in autophagy initiation genes associated with vascular phenotypes. Additionally, Dr. Lino Cardenas will learn advanced techniques in molecular biology and conditional gene deletion in multiple murine models. (3) To develop under the guidance of his mentors and advisory committee the necessary skills of directing a laboratory, fostering productive research collaborations and grant writing (4) To successfully apply for R01 funding within 3 years of his award initiation.