Development of gene editing based therapy for cardiovascular diseases

开发基于基因编辑的心血管疾病疗法

• 批准号: 10313701
• 负责人: YUQING Eugene CHEN
• 金额: $ 71.73万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10313701
• 关键词: Address; Animal Model; Animals; Antibodies; Aromatase; Atherosclerosis; Award; CRISPR/Cas technology; Cardiovascular Diseases; Cardiovascular system; Cause of Death; Cells; Chemistry; Cholesterol; Clinical Treatment; Clinical Trials; Combined Modality Therapy; Coronary; Coronary Arteriosclerosis; DNA; DNA Double Strand Break; Dangerousness; Dependovirus; Development; Diet; Disease; Drug Targeting; Dyslipidemias; Elements; Engineering; Enzymes; Evaluation; Event; Frequencies; Genes; Genetic; Genome; Hepatocyte; High Density Lipoproteins; High Fat Diet; Human; Hyperlipidemia; Hypertriglyceridemia; In Vitro; Incidence; Knock-out; Knowledge; LDLR gene; Lipids; Longevity; Low-Density Lipoproteins; Mediating; Meta-Analysis; Methods; Modeling; Monitor; Mus; Mutation; New Zealand; Nobel Prize; Oryctolagus cuniculus; Patients; Pharmaceutical Preparations; Pharmacologic Substance; Positioning Attribute; Pre-Clinical Model; Proteins; RNA Interference; Reporting; Residual state; Risk; Safety; Sagittaria; Study models; Therapeutic; Transcript; Triglycerides; Tumorigenicity; Variant; Very low density lipoprotein; Work; adeno-associated viral vector; apolipoprotein C-III; atherogenesis; atherosclerosis risk; base; cardiovascular disorder prevention; cardiovascular disorder risk; cardiovascular disorder therapy; density; efficacy evaluation; experimental study; gene therapy; genotoxicity; hypercholesterolemia; improved; in vivo; infancy; inhibitor/antagonist; insight; intein; knockout gene; lipid metabolism; mouse model; new therapeutic target; novel; novel therapeutics; response; translational study

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.

项目概要/摘要 最近的治疗进展大大降低了心血管疾病（CVD）的发病率。这 低密度（LDL）和极低密度（VLDL）脂蛋白的作用在动脉粥样硬化形成中至关重要。 无论过去30年临床治疗取得怎样的进展，很大一部分接受他汀类药物治疗的患者 患者，无论是否接受联合治疗，LDL-C降低不足且残留风险仍然较高 仍然存在，可能与持续相对较高的甘油三酯（TG）水平有关，从而限制了 这些疗法。这强调需要针对 CVD 脂质代谢的其他新疗法 预防和治疗。在本提案中，我们建议通过开发 ApoC3 的遗传缺陷 通过 Cas9 碱基编辑器 (Cas9-BE) 策略，腺相关病毒 (AAV) 介导 ApoC3 体内沉默 (AAV-Cas9-BE-ApoC3) 提供保护，防止高胆固醇饮食引起的高胆固醇血症和 兔子的动脉粥样硬化。具体来说，我们将 1) 在临床前模型物种中开发 AAV-Cas9-BE-ApoC3， 新西兰白兔。我们将利用体外培养的兔确定最佳靶向策略 细胞，然后通过实验确定最佳递送参数以实现有效的 ApoC3 基因 兔肝细胞的敲除； 2) 使用最佳条件评估 AAV-Cas9-BE-ApoC3 的功效 目标 1 确定敲除兔子中的 ApoC3； 3）对AAV的安全性进行多年评估- 兔子体内的 Cas9-BE-ApoC3。通过利用新的 CRISPR/Cas9 技术来靶向完成这些目标 ApoC3 将提供令人信服的证据，将 ApoC3 确立为基于基因编辑的新型可行靶标 治疗高脂血症和CVD。