Genome Editing Treatment for Catecholaminergic Polymorphic Ventricular Tachycardia

儿茶酚胺能多形性室性心动过速的基因组编辑治疗

基本信息

批准号：
10441135
负责人：
Oliver Moore
金额：
$ 4.71万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-03-01 至 2025-09-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10441135
关键词：
3-Dimensional Address Affect Age Alleles Arrhythmia CRISPR therapeutics CRISPR/Cas technology Calcium Calcium Channel Cardiac Cardiac Myocytes Cardiovascular system Catecholaminergic Polymorphic Ventricular Tachycardia Confocal Microscopy Data Defibrillators Dependovirus Disease Double Effect Echocardiography Effectiveness Electric Stimulation Electrocardiogram Emotional Stress Event Exertion Genes Genetic Goals Heart Heart Abnormalities Heart Diseases Human Human Genome Image Individual Inherited Injections Knowledge Lead Left Lentivirus Light Long-Term Effects Measures Medical Mendelian disorder Messenger RNA Methods Microscopy Mission Molecular Analysis Mus Muscle Cells Mutation Operative Surgical Procedures Organoids Outcome Patients Pharmacological Treatment Physiological Pre-Clinical Model Predisposition Protein Isoforms Proteins Public Health Reporter Genes Research Research Project Grants Risk RyR2 Ryanodine Receptor Calcium Release Channel Sarcoplasmic Reticulum Silent Mutation Site Specificity Stress Tests Sudden Death Sympathectomy Syncope System Technology Testing United States National Institutes of Health Variant Ventricular Ventricular Arrhythmia Ventricular Tachycardia base editing causal variant confocal imaging deep sequencing disability disease-causing mutation effective therapy gene therapy genome editing heart function heart rhythm human disease implantation improved in vivo induced pluripotent stem cell induced pluripotent stem cell derived cardiomyocytes insertion/deletion mutation knock-down metabolomics mortality mouse model mutant mutation correction non-compliance novel novel strategies novel therapeutic intervention preservation prevent prime editing repaired side effect sudden cardiac death targeted treatment therapeutic evaluation therapeutic genome editing vector

项目摘要

There is a fundamental gap in treatment for Catecholaminergic Polymorphic Ventricular Tachycardia (CPVT). Pharmacological treatment of CPVT is partially effective for a disease that causes sudden death. A fuller understanding of novel gene editing approaches is needed to develop safe, non-surgical, and effective therapies that target the underlying causes of CPVT. In approximately 60% of CPVT patients, mutations in RYR2 cause arrhythmia through abnormal calcium handling in cardiomyocytes. The lab has shown that using CRISPR/Cas9 to knockdown the RYR2 mutant allele is effective in preventing ventricular tachycardia(VT) by decreasing expression of dysfunctional RyR2. The overall objective of this application is to improve therapy for patients with CPVT. Preliminary data generated by the applicant showed that directly targeting a disease- causing mutation with traditional CRISPR/Cas9 therapy in a CPVT mouse model prevented pacing induced VT but reduced total expression of RyR2. The central hypothesis of this proposal is that novel methods of CRISPR/Cas9 gene editing can treat CPVT by specifically correcting causative mutation sites, reducing mutant RyR2 expression while preserving total RyR2 expression, normalizing Ca2+ handling, and decreasing susceptibility to VT. The rationale for this research is that an understanding of the effectiveness and specificity of gene editing in the correcting mutations in the heart may lead to safe novel approaches to treat genetic cardiac disorders. The hypothesis will be tested with the following specific aims 1) test the therapeutic potential of human RYR2 mutation correction in an iPSC-CM preclinical model of CPVT and 2) test the therapeutic potential of mouse RYR2 mutation correction in a mouse model of CPVT. To determine aim 1, we deliver CRISPR/Cas9 prime editing vectors to IPSC, derive CM, and measure function through confocal and light sheet imaging of IPSC and 3D cardiac organoids. To determine aim 2, we will use Lenti-CRISPR/Cas9 prime editing vectors to treat mouse models of CPVT and measure long-term cardiac function through echocardiography, EKG, programmed electrical stimulation, and molecular analysis. This research is significant in that it will advance gene editing correction of RYR2 mutations as a safe and effective method for the treatment of CPVT.

儿茶酚胺能多态性心脏心动过速（CPVT）的治疗方面存在基本差距。 CPVT的药理学治疗对于导致猝死的疾病部分有效。一个饱满需要了解新型基因编辑方法来发展安全，非手术和有效针对CPVT根本原因的治疗方法。在大约60％的CPVT患者中，突变 RYR2通过心肌细胞中的异常钙处理引起心律不齐。实验室表明使用 CRISPR/CAS9敲除RyR2突变等位基因可有效防止心室心动过速（VT）功能失调的RYR2的表达降低。该应用的总体目的是改善 CPVT患者。申请人产生的初步数据表明，直接针对疾病 - 在CPVT小鼠模型中使用传统CRISPR/CAS9治疗引起突变，阻止了起搏引起的VT 但是降低了RYR2的总表达。该提议的中心假设是 CRISPR/CAS9基因编辑可以通过特异性纠正因果突变位点来治疗CPVT，从而减少突变体 RYR2表达在保留总RYR2表达，标准化Ca2+处理和减少的同时对VT的敏感性。这项研究的理由是了解有效性和特异性心脏校正突变中的基因编辑可能会导致安全的新方法治疗遗传心脏疾病。该假设将以以下特定目的进行检验1）测试治疗潜力 CPVT的IPSC-CM临床前模型中的人RYR2突变校正和2）测试治疗小鼠RYR2突变校正的电位在CPVT的小鼠模型中。为了确定目标1，我们交付 CRISPR/CAS9素数编辑向量iPSC，得出CM并通过共焦和光线量 IPSC和3D心脏器官的薄片成像。要确定目标2，我们将使用lenti-crispr/cas9 Prime 编辑向量以治疗CPVT的小鼠模型并通过超声心动图，心电图，编程的电刺激和分子分析。这项研究是重要的是，它将推动基因编辑校正RYR2突变，作为一种安全有效的方法 CPVT的处理。