Mechanobiology of Endothelial-to-Mesenchymal Transition in Cardiovascular Calcification

摘要 这份提案描述了一个为期五年的有指导的医生-科学家培训计划，以描述 心血管钙化中内皮间充质转化(EndMT)的机制生物学。 心血管钙化是一种非常普遍的过程，在这种过程中，血管内形成钙矿物。 和心脏组织，它的存在与发病率和死亡率的增加有关。尽管 几十年的研究，仍然没有有效的药物疗法来治疗这一过程。最近的工作有 EndMT与心血管钙化有关。EndMT描述了内皮细胞 心脏瓣膜或血管的内壁被刺激去分化为间充质细胞。 随后，这些间充质细胞能够分化为各种谱系，包括 可促进钙化的软骨和成骨谱系。虽然EndMT是一个很有希望的目标 心血管钙化的治疗干预：实验性EndMT过程的特征 学习可能是具有挑战性的。目前评估EndMT的方法，如遗传谱系追踪研究和 免疫染色检测内皮和间质标记物的表达，是劳力、资源和时间- 密集的。此外，这些方法是对EndMT的二元评估，并未涵盖 这个过程--它的动态机械性质。接受EndMT治疗的内皮细胞改变其细胞骨架 力学，脱离其邻近的细胞，并迁移到间隙空间。因此，他们的机械 在EndMT过程中，特性或“机械表型”可能是动态的，但以前从未被探索过。 在这项建议中，我们概述了我们的目标是表征EndMT的机械表型，希望 开发新的高通量平台，以更快地确定EndMT的可能治疗靶点- 相关疾病，包括心血管钙化。此外，随之而来的是指导职业发展 培训计划将允许应聘者发展成功完成该项目所需的专业知识，并 将为他提供必要的指导和支持，使他成为一名完全独立的科学调查员。 根据NHLBI的战略目标，这项建议的目的集中在：(1)澄清我们的理解 EndMT和心血管钙化的病理生物学研究，以及(2)开发创新的新平台以 加快我们确定这些过程的治疗靶点的能力，从而推动 研究。

ABSTRACT This proposal describes a five-year mentored physician-scientist training program to characterize the mechanobiology of endothelial-to-mesenchymal transition (EndMT) in cardiovascular calcification. Cardiovascular calcification is a highly prevalent process whereby calcium mineral forms within the blood vessels and heart tissue, and its presence is associated with an increased risk of morbidity and mortality. Despite decades of research, there remains no effective medical therapy to treat this process. Recent work has implicated EndMT in cardiovascular calcification. EndMT describes the phenomenon whereby endothelial cells lining the lumen of the heart valves or blood vessels are stimulated to dedifferentiate into mesenchymal cells. Subsequently, these mesenchymal cells are capable of differentiating into various lineages, including the chondrogenic and osteogenic lineages that can promote calcification. While EndMT is a promising target for therapeutic intervention in cardiovascular calcification, characterizing the EndMT process in experimental studies can be challenging. Current methods to assess EndMT, such as genetic lineage tracing studies and immunostaining detection of endothelial and mesenchymal marker expression, are labor-, resource-, and time- intensive. Further, these methods are binary assessments of EndMT and do not capture an important aspect of the process – its dynamic mechanical nature. Endothelial cells undergoing EndMT alter their cytoskeletal mechanics, detach from their neighboring cells, and migrate into the interstitial space. Thus, their mechanical properties, or “mechanophenotype,” during EndMT are likely dynamic, but have not been previously explored. In this proposal, we outline our aims to characterize the mechanophenotypes of EndMT, with the hope of developing novel, high-throughput platforms to more rapidly identify possible therapeutic targets for EndMT- related disease, including cardiovascular calcification. Further, the accompanying mentored career development training plan will allow the candidate to develop the expertise needed to successfully complete this project and will provide him with the mentorship and support necessary to become a fully independent scientific investigator. In line with the strategic goals of the NHLBI, the aims of this proposal focus on: (1) elucidating our understanding of the pathobiology of EndMT and cardiovascular calcification, and (2) developing innovative new platforms to accelerate our ability to identify therapeutic targets for these processes, thereby advancing translational research.