Use of Tissue Engineered Heart Valves to study Calcific Aortic Valve Disease

项目摘要 钙化性主动脉瓣病(CAVD)是指主动脉瓣逐渐僵硬和钙化， 作为含氧血液从左心室单向流动到主动脉的通道 心跳。在严重的CAVD病例中，瓣膜无法打开以允许血液从心脏流向身体， 这可能会对左心室造成严重损害，导致心力衰竭和死亡。尽管它的高 CAVD的流行，到目前为止还没有有效的药物治疗方法，迫使患者接受 心脏手术接受救命治疗。我们认为，对生物多样性的动态阶段缺乏了解 早期、中期和晚期疾病与对异质性分化性疾病的有限了解相结合 细胞群体和体外药物测试平台的使用没有捕捉到疾病的全部复杂性 这是迄今为止缺乏临床有效的药物疗法的原因之一。 为了解决CAVD领域的这些关键空白，我将首先将我们实验室的组织工程机械结合起来 主动共培养瓣膜模型与先进的实时OCT成像，以获取3D时间推移图像 钙化过程。接下来，我将利用我们的组织工程瓣膜模型的单细胞测序来揭示 异质性、易发病和抗病的瓣膜细胞的特征。最后，我将结合 从多维药物测试项目的前两个阶段产生的信息和技术 研究以前发现的药物靶点NFkB途径何时以及为什么， 减少体外钙化。 在本项目结束时，我们将发现ECM中前所未有的延时阶段，细胞 对瓣膜钙化至关重要的形态和矿化变化以及分子途径 与诱导或防止钙化的异质细胞群有关。我们将利用这些 共同评估一个有前景的治疗靶点的进展不仅是因为它是否有效，而且是因为它 什么时候和为什么它对3D体外瓣膜矿化有影响。药品检测平台作为一种 这一项目的结果可用于严格评估我们的其他有希望的治疗靶点 单细胞测序结果和其他实验室的测序结果，允许更快速、更具成本效益地评估CAVD 药物治疗学。

Project Summary Calcific Aortic Valve Disease (CAVD) is the gradual stiffening and calcification of the aortic heart valve, which serves as a gateway for unidirectional flow of oxygenated blood from the left ventricle to the aorta during each heartbeat. In severe cases of CAVD, the valve cannot open to allow blood to travel from the heart to the body, which can cause severe damage to the left ventricle resulting in heart failure and death. Despite its high prevalence, there are no effective pharmacological therapeutics to date for CAVD, forcing patients to undergo heart surgery to receive life-saving treatment. We believe that a lack of knowledge about the dynamic stages of early, mid, and late disease combined with a limited understanding of the heterogenous differentiated diseased cell populations and the use of in-vitro drug testing platforms that do not capture the full complexity of the disease contribute to this lack of clinically effective pharmacological therapeutics to date. To address these critical gaps in the CAVD field, I will first combine our lab’s tissue engineered mechanically active co-culture valve model with cutting-edge live-OCT imaging to acquire 3D time-lapse images of the calcification process. Next, I will utilize single-cell sequencing of our tissue-engineered valve model to uncover characteristics of heterogeneous disease-prone and disease-resistant valvular cells. Finally, I will combine the information and technologies produced from the first two stages of the project in a multi-dimensional drug testing platform to investigate when and why a previously discovered pharmacological drug target, NFkB pathway, reduces calcification in-vitro. At the conclusion of this project, we will uncover never-before seen time-lapse stages in ECM, cellular morphology and mineralization changes that are critical for valvular calcification, as well as molecular pathways related to heterogenous cell groups that are inductive or preventative of calcification. We will utilize these advancements together to evaluate a promising therapeutic target not just for whether it has an effect, but rather when and why it has an effect on 3D in-vitro valvular mineralization. The drug-testing platform produced as a result of this project may be utilized to rigorously evaluate other promising therapeutic targets identified by our single-cell sequencing results and by other labs, allowing for more rapid and cost-effective evaluation of CAVD pharmacological therapeutics.