Mechanobiology of Endothelial-to-Mesenchymal Transition in Cardiovascular Calcification

摘要 该提案描述了一个为期五年的指导医生科学家培训计划，以描述 心血管钙化中内皮-间质转化（EndMT）的机械生物学。 心血管钙化是一个非常普遍的过程，其中钙矿物质在血管内形成 和心脏组织，并且其存在与发病率和死亡率的增加的风险相关。尽管 经过几十年的研究，仍然没有有效的医学疗法来治疗这一过程。最近的工作已经 提示EndMT参与心血管钙化。EndMT描述了内皮细胞 心脏瓣膜或血管内腔的衬里被刺激去分化成间充质细胞。 随后，这些间充质细胞能够分化成各种谱系，包括骨髓间充质干细胞。 成软骨和成骨谱系，可以促进钙化。虽然EndMT是一个很有前途的目标， 心血管钙化的治疗干预，表征实验性EndMT过程， 研究可能是具有挑战性的。当前评估EndMT的方法，例如遗传谱系追踪研究和 免疫染色检测内皮和间充质标志物的表达，是劳动力，资源，和时间- 密集。此外，这些方法是EndMT的二元评估，并且没有捕获EndMT的重要方面。 这个过程-它的动态机械性质。内皮细胞接受EndMT改变其细胞骨架 机械，脱离其邻近细胞，并迁移到间隙空间。因此，他们的机械 EndMT过程中的性质或“机械表型”可能是动态的，但以前没有探索过。 在这个建议中，我们概述了我们的目标，以表征EndMT的机械表型，希望 开发新的高通量平台，以更快地识别EndMT的可能治疗靶点， 相关疾病，包括心血管钙化。此外，伴随而来的指导性职业发展 培训计划将允许候选人发展成功完成该项目所需的专业知识， 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《无法律约束力文书》的战略目标，本提案的目标侧重于：（1）阐明我们的理解 EndMT和心血管钙化的病理生物学，以及（2）开发创新的新平台， 加快我们确定这些过程的治疗靶点的能力，从而促进翻译 research.

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.