Development of gene editing based therapy for cardiovascular diseases

项目总结/摘要 最近的治疗进展大大降低了心血管疾病（CVD）的发病率。的 低密度脂蛋白（LDL）和极低密度脂蛋白（VLDL）的作用在动脉粥样硬化形成中是关键的。 无论过去30年临床治疗的进展如何，对于很大比例的他汀类药物治疗的患者， 接受或不接受联合治疗的患者，LDL-C降低不充分，剩余风险仍相对较高 仍然存在，可能与持续相对较高的甘油三酯（TG）水平相关，从而限制了 这些疗法。这强调了需要额外的新的治疗靶向血脂代谢的心血管疾病 预防和治疗。在这项提案中，我们建议通过以下途径发展ApoC 3的遗传缺陷： 腺相关病毒（AAV）通过Cas9碱基编辑器（Cas9-BE）策略介导的ApoC 3的体内沉默 （AAV-Cas9-BE-ApoC 3）以提供针对高胆固醇饮食诱导的高胆固醇血症的保护，以及 家兔动脉粥样硬化。具体地，我们将1）在临床前模型物种中开发AAV-Cas9-BE-ApoC 3， 新西兰白色兔子。我们将使用体外培养的兔来确定最佳的靶向策略 细胞，然后进行实验以确定最佳递送参数，以实现有效的ApoC 3基因 2）使用最佳条件评估AAV-Cas9-BE-ApoC 3的功效 在目的1中确定的AAV-1基因敲除兔中的ApoC 3; 3）对AAV-1基因敲除兔中的ApoC 3的安全性进行多年评估。 兔中的Cas9-BE-ApoC 3。通过利用新的CRISPR/Cas9技术来实现这些目标， ApoC 3将为建立ApoC 3作为基于基因编辑的新的可行靶点提供令人信服的证据。 治疗高脂血症和心血管疾病。

PROJECT SUMMARY/ABSTRACT Recent therapeutic advances considerably reduced the incidence of cardiovascular disease (CVD). The contribution of low-density (LDL) and very low density (VLDL) lipoproteins is critical in atherogenesis. Regardless the progress of clinical treatments in the past 30 years, for a significant proportion of statin-treated patients, with or without combination therapy, insufficient LDL-C reduction and relatively high residual risk still remain, likely associated with persistent relatively high triglyceride (TG) levels, thus limiting the benefits of these therapies. This underscores the need for additional new therapies targeting lipid metabolism in CVD prevention and treatment. In this proposal, we propose to develop genetic deficiency of ApoC3 through an adeno-associated virus (AAV) mediated in vivo silencing of ApoC3 by Cas9 base editor (Cas9-BE) strategy (AAV-Cas9-BE-ApoC3) to render protection against high cholesterol diet-induced hypercholesterolemia and atherosclerosis in rabbits. Specifically, we will 1) develop AAV-Cas9-BE-ApoC3 in a preclinical model species, the New Zealand White rabbits. We will determine the optimal targeting strategies using in vitro cultured rabbit cells, followed by experiments to determine the optimal delivery parameters to achieve effective ApoC3 gene knockout in rabbit hepatocytes; 2) evaluate the efficacy of AAV-Cas9-BE-ApoC3 using optimal conditions determined in Aim 1 to knockout ApoC3 in rabbits; 3) conduct multiple-year evaluation of the safety of AAV- Cas9-BE-ApoC3 in rabbits. Completion of these aims by leveraging new CRISPR/Cas9 technology to target ApoC3 will provide compelling evidence to establish ApoC3 as a novel feasible target for gene editing-based therapy for hyperlipidemia and CVD.