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

项目摘要 钙化性主动脉瓣疾病（CAVD）是主动脉心脏瓣膜的逐渐硬化和钙化， 在每一次心脏手术期间， 心跳在严重的CAVD病例中，瓣膜不能打开以允许血液从心脏流向身体， 这会对左心室造成严重损害，导致心力衰竭和死亡。尽管其高 由于CAVD的流行，迄今为止还没有有效的药物治疗方法，迫使患者接受 接受心脏手术以挽救生命。我们认为，缺乏对动态阶段的了解， 早期、中期和晚期疾病结合对异质性分化疾病的有限理解， 细胞群和使用体外药物测试平台，无法捕捉疾病的全部复杂性 导致迄今为止缺乏临床有效的药理学治疗。 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联合收割机 主动共培养瓣膜模型，采用先进的实时OCT成像，以获取瓣膜的3D延时图像。 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 异质性易患病和抗病瓣膜细胞的特征。最后，我将联合收割机 从项目的前两个阶段产生的信息和技术在一个多维的药物测试 平台，以调查何时以及为什么以前发现的药理学药物靶点，NF κ B途径， 减少体外钙化。 在这个项目的结论，我们将发现以前从未见过的时间推移阶段，在ECM，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 与诱导或预防钙化的异质细胞群有关。我们将利用这些 一起评估一个有前途的治疗目标，不仅是为了它是否有效果， 何时以及为什么它对3D体外瓣膜矿化有影响。该药物测试平台作为一个 该项目的结果可用于严格评估我们确定的其他有前途的治疗靶点。 单细胞测序结果和其他实验室，允许更快速和更具成本效益的CAVD评估 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.