Creating Engineered Models of Valvular Disease to Study Anti-Calcific Therapies

描述(申请人提供)：3-羟基-3-甲基戊二酰辅酶A还原酶抑制剂，或他汀类药物，以其降低胆固醇的能力而闻名，是世界上处方最多的药物。尽管他汀类药物在抑制动脉粥样硬化进展方面具有无可争辩的积极作用，但最近关于他汀类药物对钙化性瓣膜狭窄的影响存在很大争议，钙化性瓣膜狭窄是最常见的心脏瓣膜疾病类型。由于许多因素，包括他汀类药物在全身的多效性(通常是自相矛盾的)效应，为了阐明他汀类药物对心脏瓣膜组织和功能的影响，很有必要进行体外对照研究。因此，为了：(1)更好地了解瓣膜钙化的进展，(2)开发更多预防和治疗瓣膜钙化的方法，以及(3)更好地了解世界上最常见的处方药如何影响(以及可能有益于)心脏瓣膜，我们的目标是建立瓣膜疾病的体外和体外模型，并在对照实验中使用这些模型来探索和表征他汀类药物对瓣膜功能的影响。目的1：研究他汀类药物对不同成分体外培养的瓣膜间质细胞(VIC)功能的影响。在目标1中，来自健康和疾病来源的VIC将主要在确定成分的二维环境中培养，用不同剂量的他汀类药物治疗不同时间，然后检查钙化/功能障碍的指标。这些实验的结果将使我们了解他汀类药物如何调节VIC功能障碍，培养成分在调节VIC对他汀类药物的反应中的作用，以及他汀类药物的限制/能力。目的2：建立和表征心脏瓣膜病的三维模型。利用体外器官和体外细胞培养，我们将通过多种不同的方法来制作与生理相关的瓣膜病变模型。我们量身定做的瓣膜病变环境将为研究天然瓣膜疾病的进展和各种因素对瓣膜功能的影响提供一个受控和容易获得的培养系统。目的：利用体外和体外心脏瓣膜病三维模型，评价他汀类药物抗钙化治疗方案的疗效。在目标3中，我们将结合从目标1和目标2中获得的知识，将目标1中确定的他汀类药物治疗方案应用于目标2中创建的“合成疾病”瓣膜模型。通过这些实验，我们将能够评估他汀类药物在生理相关环境中的能力和局限性，分析他汀类药物作用的中间阶段，并最终验证我们的模型作为抗钙化药物体外测试的平台是否有效。这项研究的结果可能具有重大的临床意义，因为拟议的工作满足了临床心血管修复的几个关键需求，例如更好地了解瓣膜疾病，探索治疗或抑制瓣膜狭窄的潜在疗法。这项拟议工作的目标是研究他汀类药物如何影响心脏瓣膜细胞的功能。他汀类药物是全球数百万人服用的降胆固醇药物。这些实验直接适用于医学和公共卫生，因为它们将产生数据，使人们能够更好地了解心脏瓣膜疾病的原因以及他汀类药物与心脏瓣膜相互作用的机制。我们的研究还将产生钙化性瓣膜疾病的体外模型，可用于测试其他抗钙化药物或策略。

DESCRIPTION (provided by applicant): 3-hydroxy-3-methyglutaryl-coenzyme A reductase inhibitors, or statins, are best known for their cholesterol-lowering abilities, and are the most prescribed drugs in the world. While statins have indisputably positive effects on inhibiting the progression of atherosclerosis, much controversy has recently arisen regarding the impact of statins on calcific valvular stenosis, the most common type of heart valve disease. Due to many factors, including the pleiotropic (and often paradoxical) effects of statins throughout the body, controlled in vitro studies are greatly needed in order to elucidate the effect of statins on heart valve tissue and function. Thus, in order to: (1) better understand the progression of valve calcification, (2) develop more options for prevention and treatment of valve calcification, and (3) better understand how the most prescribed drug in the world affects (and possibly benefits) heart valves, we aim to create ex vivo and in vitro models of valvular disease, and use these models in controlled experiments to explore and characterize the effects of statins on valve function. The following specific aims will help us to achieve this goal: AIM 1: Characterize the effects of statin treatment on valvular interstitial cell (VIC) (dys)function in in vitro cultures with varying compositions. In Aim 1, VICs from both healthy and diseased sources will be cultured in primarily 2-D environments of defined compositions, treated with different dosages of statins for varying lengths of time, and then examined for indicators of calcification/dysfunction. The results from these experiments will allow us to understand how statins regulate VIC dysfunction, the role of culture composition in regulating VIC response to statins, and the limitations/capabilities of statins. AIM 2: Create and characterize 3-D models of valvular disease. Using ex vivo organ and in vitro cell cultures, we will proceed through multiple different approaches to produce physiologically relevant models of diseased valves. Our tailored diseased valve environments will offer a controlled and readily-available culture system for studying the progression of native valve disease and the effects of various agents on valve function. AIM 3: Using 3-D ex vivo and in vitro models of valvular disease, evaluate the efficacy of statin- based anti-calcification treatment regimens. In Aim 3, we will combine the knowledge gained from Aims 1 and 2 by applying the statin treatment regimens identified in Aim 1 to the `synthetically-diseased' valve models created in Aim 2. Through these experiments, we will be able to evaluate statin capabilities and limitations in physiologically-relevant environments, analyze intermediate stages of statin action, and, ultimately, validate whether our models are effective as platforms for in vitro testing of anti-calcific drugs. The results of this investigation are likely to have significant clinical implications, as the proposed work addresses several critical needs in clinical cardiovascular repair, such as gaining a better understanding of valvular disease and exploring potential therapies to treat or inhibit valvular stenosis. The goal of the proposed work is to study how statins, cholesterol-lowering drugs which are taken by millions of people worldwide, affect the function of heart valve cells. These experiments are directly applicable to medicine and public health in that they will generate data that will enable a better understanding of the causes of heart valve disease and the mechanism of how statins interact with heart valves. Our study will also generate in vitro models of calcific valvular disease that may be used for testing other anti-calcification pharmaceuticals or strategies.