Gene Therapy Design Principles for Duchenne Muscular Dystrophy

杜氏肌营养不良症的基因治疗设计原则

• 批准号: 10723951
• 负责人: Asuka Eguchi
• 金额: $ 16.2万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10723951
• 关键词: Address; Advisory Committees; Affect; Award; Biochemical; Biochemistry; Biological Assay; Biomedical Engineering; Calcium; Cardiac; Cardiac Myocytes; Cardiomyopathies; Cardiovascular Diseases; Cardiovascular system; Cell Death; Cell Survival; Cell membrane; Cells; Cellular Morphology; Cellular biology; Cicatrix; Code; Contracts; Cytoskeletal Proteins; Data; Defect; Dependovirus; Development Plans; Dilatation - action; Dilated Cardiomyopathy; Disease; Disease Progression; Disease model; Duchenne cardiomyopathy; Duchenne muscular dystrophy; Dyes; Dystrophin; Echocardiography; Engineering; Ensure; Exhibits; Extracellular Matrix; Fibrosis; Functional disorder; Heart; Heart Diseases; Heart failure; Histology; Human; Hydrogels; Image; Impairment; Learning; Left ventricular structure; Length; Life; Longevity; Luciferases; Measures; Mendelian disorder; Mentored Research Scientist Development Award; Mentors; Mentorship; Methods; Microscopy; Muscle; Muscular dystrophy cardiomyopathy; Mutation; Onset of illness; Pathogenicity; Pathologic; Patients; Phenotype; Proteins; Quality of life; Reactive Oxygen Species; Reporter; Research; Research Personnel; Respiratory Failure; Skeletal Muscle; Stroke Volume; Symptoms; Techniques; Telomere Shortening; Testing; Therapeutic Effect; Thinness; Tissues; Traction Force Microscopy; Training; Variant; Ventricular; Western Blotting; adeno-associated viral vector; career; career development; design; efficacy evaluation; experimental study; gene therapy; heart function; improved; induced pluripotent stem cell; innovation; link protein; lipid nanoparticle; male; mouse model; muscle degeneration; mutation correction; nanoparticle delivery; novel; pediatric cardiologist; premature; prevent; programs; promoter; protein expression; ratiometric; reduce symptoms; response; restoration; skeletal muscle wasting; skills; telomere; therapy design; vector

Project Summary/Abstract The long-term objective of this study is to develop gene therapies that treat Duchenne muscular dystrophy (DMD) cardiomyopathy. DMD cardiomyopathy, characterized by ventricular chamber enlargement and thinning of the ventricular wall, ultimately leads to heart failure. Pathogenic features of DMD cardiomyocytes include contractile dysfunction, poor calcium handling, elevated reactive oxygen species, telomere shortening, and premature cell death. When a large number of cells die in the heart, scar tissue forms, increasing the stiffness of the heart. Although there are treatments available to alleviate symptoms of dilated cardiomyopathy, there are currently no therapies to prevent or delay the onset of this disease. Smaller versions of dystrophin amenable to gene therapy have shown promise to treat DMD-associated severe skeletal muscle wasting; however, surprisingly little is known about their effects in treating heart failure. This research plan will leverage bioengineered hydrogels of tunable stiffness, human induced pluripotent stem cells (iPSCs) with dystrophin mutations, and biochemical techniques to determine if full-length dystrophin can rescue DMD cardiomyocytes from their pathogenic demise. During the K01 award period, Dr. Asuka Eguchi will train under the mentorship of Dr. Helen Blau, an expert on DMD. By engineering hydrogels that mimic stiff, diseased heart tissue, Dr. Eguchi will be able to measure parameters of contraction in cardiomyocytes differentiated from DMD iPSCs. Aim 1 will test if full-length dystrophin can rescue DMD cardiomyocytes from contractile deficits, aberrant calcium handling, and premature cell death. Aim 2 will determine if split vector or lipid nanoparticle approaches can deliver full-length dystrophin to cardiomyocytes. Aim 3 will test whether this gene therapy strategy to deliver full-length dystrophin can delay the onset of DMD cardiomyopathy in a mouse model. Gene therapy approaches targeting the root cause of disease, the lack of dystrophin, is critical for extending lifespan and improving the quality of life of DMD patients. The career development plan is designed to enable Dr. Eguchi to successfully transition to a career as independent investigator. Her scientific advisory committee consist of Dr. Beth Pruitt, a bioengineer with expertise in traction force microscopy, Dr. Joseph Wu, an expert on cardiovascular disease modeling, and Dr. Daniel Bernstein, a pediatric cardiologist. Collectively, these collaborators will help Dr. Eguchi develop skills at the interface of bioengineering, cell biology, and biochemistry to launch an independent research program in cardiovascular research.

项目总结/摘要 本研究的长期目标是开发治疗杜氏肌营养不良症（DMD）的基因疗法 心肌病DMD心肌病，以心室腔扩大和心室壁变薄为特征， 心室壁，最终导致心力衰竭。DMD心肌细胞的致病特征包括收缩性 功能障碍、钙处理不良、活性氧升高、端粒缩短和早熟细胞 死亡当大量的细胞在心脏中死亡时，疤痕组织形成，增加心脏的硬度。 虽然有治疗方法可以缓解扩张型心肌病的症状，但目前还没有 预防或延缓这种疾病的发作。适合基因治疗的较小版本的抗肌萎缩蛋白 已经显示出治疗DMD相关的严重骨骼肌萎缩的前景;然而，令人惊讶的是， 已知它们在治疗心力衰竭中的作用。这项研究计划将利用生物工程水凝胶， 可调刚度，具有肌营养不良蛋白突变的人诱导多能干细胞（iPSC），以及生物化学 技术，以确定是否全长肌营养不良蛋白可以拯救DMD心肌细胞从他们的致病性死亡。 在K 01奖励期间，江口明日香博士将在海伦·布劳博士的指导下进行培训，海伦·布劳博士是一位专家， DMD。通过设计模拟僵硬的患病心脏组织的水凝胶，江口博士将能够测量 图4示出了从DMD iPSC分化的心肌细胞中的收缩参数。目标1将测试是否全长 肌营养不良蛋白可以挽救DMD心肌细胞的收缩缺陷，异常钙处理，和过早 细胞死亡目标2将确定是否分裂载体或脂质纳米颗粒的方法可以提供全长肌营养不良蛋白 到心肌细胞。目的3将测试这种基因治疗策略，以提供全长肌营养不良蛋白是否可以延迟 DMD心肌病的发病机制基因治疗方法针对的根本原因 在DMD患者中，缺乏肌营养不良蛋白对于延长DMD患者的寿命和改善其生活质量至关重要。 职业发展计划旨在使江口博士能够成功地过渡到职业生涯， 独立调查员她的科学顾问委员会由Beth Pruitt博士组成，她是一位生物工程师， 在牵引力显微镜的专业知识，约瑟夫吴博士，心血管疾病建模专家，和博士。 丹尼尔伯恩斯坦，儿科心脏病专家。总的来说，这些合作者将帮助江口博士发展技能， 生物工程，细胞生物学和生物化学的接口，以启动一个独立的研究计划， 心血管研究