Evaluation of adeno-associated viral (AAV) mediated gene replacement therapy as a therapeutic option for SLC25A4 deficiency

评估腺相关病毒 (AAV) 介导的基因替代疗法作为 SLC25A4 缺陷的治疗选择

基本信息

批准号：
10606065
负责人：
Abigail Benkert
金额：
$ 7.63万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10606065
关键词：
Adrenergic Agents Affect Base Pairing Biochemical Cardiac Myocytes Cardiomyopathies Cell Line Cell model Cells Codon Nucleotides Complementary DNA DNA Data Development Disease Disease Progression Evaluation Exertion Family Fellowship Foundations Frameshift Mutation Functional disorder Generations Goals Heart Transplantation Heart failure Hereditary Disease Hypertrophic Cardiomyopathy Impairment In Vitro Individual Inherited Inner mitochondrial membrane Laboratories Lactic Acidosis Link Mediating Mendelian disorder Mennonite Mitochondria Mitochondrial Proteins Modality Modeling Modification Molecular Muscle Cells Muscle Fibers Mutation Myocardial Myocardial dysfunction Myocardium Myopathy Natural History Nuclear Organ Organoids Oxidative Phosphorylation Patients Phenotype Pre-Clinical Model Protein Isoforms Proteins Recombinants Recovery Research Research Personnel SLC25A4 gene Scientist Secondary to Skeletal Development Skeletal Muscle Structure Surgeon Systemic Therapy Therapeutic Tissues Training Transgenes Translating Translational Research Translations Tropism Viral Viral Vector autosome career cohort delivery vehicle disease phenotype efficacy evaluation experimental study gene replacement gene replacement therapy genetic pedigree improved in vitro Model induced pluripotent stem cell loss of function member mitochondrial metabolism next generation null mutation premature prevent reduce symptoms restoration skeletal skills solute therapeutic gene treatment strategy vector vector biodistribution

项目摘要

Project Summary/Abstract Disruption of mitochondrial oxidative phosphorylation (OXPHOS) is associated with the development of biochemical alterations that typically affect tissues with a high energy demand, particularly skeletal and cardiac muscle. An inherited autosomal recessive skeletal myopathy and hypertrophic cardiomyopathy has been linked to loss of function of a nuclear DNA-encoded mitochondrial protein, due to a frameshift mutation in solute carrier family 25, member 4 (SLC25A4; c.523delC, p.Q175RfxX38). SLC25A4 encodes the heart-muscle isoform of the adenine nucleotide translocator-1 (ANT1, SLC25A4), which in the wild-type state is a critical component of mitochondrial metabolism. Patients with SLC25A4 deficiency display lactic acidosis, persistent adrenergic activation, and exertional intolerance secondary to both a general skeletal muscle myopathy as well as a hypertrophic cardiomyopathy. Ultimately, myocardial thickening and cardiac dysfunction progress to end- stage heart failure necessitating cardiac transplantation. There are not currently any disease-modifying therapies available for this patient cohort. However, adeno- associated viral (AAV) mediated gene replacement therapies have emerged as a powerful strategy for disease modification of inherited monogenic disorders. The long-term goal of our research is to develop a therapeutic gene replacement strategy to treat SLC25A4 deficiency. The objective of this proposal is to further characterize the disease phenotype as well as to synthesize and evaluate the efficacy of a recombinant AAV (rAAV) vector in an in vitro model of patient-derived cell lines and organoid models. The central hypothesis of this proposal is that AAV-mediated gene replacement can ameliorate the biochemical and functional effects of SLC25A4 deficiency and can more decisively prevent disease progression. The specific aims of this proposal are: 1. Characterize the SLC25A4 deficiency phenotype in patient-derived cell lines. 2. Synthesize a recombinant AAV vector for delivery of codon-optimized SLC25A4 cDNA to skeletal and cardiac myocytes. 3. Evaluate the efficacy of AAV-SLC25A4 viral transduction in patient-derived cell lines. These experiments will improve our understanding of the molecular mechanisms underlying SLC25A4 deficiency as well as allow us to evaluate the efficacy of an AAV platform in a relevant preclinical model. Moreover, the skills I will acquire during this fellowship will help to establish me as an independent investigator and a surgeon-scientist focused on the development of translational gene replacement therapies.

项目概要/摘要线粒体氧化磷酸化 (OXPHOS) 的破坏与通常影响能量需求高的组织的生化改变，特别是骨骼和心脏肌肉。遗传性常染色体隐性骨骼肌病与肥厚性心肌病存在关联由于溶质中的移码突变，核 DNA 编码的线粒体蛋白功能丧失载体家族 25，成员 4（SLC25A4；c.523delC，p.Q175RfxX38）。 SLC25A4 编码心肌腺嘌呤核苷酸易位子-1（ANT1、SLC25A4）的亚型，在野生型状态下是一个关键线粒体代谢的组成部分。 SLC25A4 缺乏症患者表现出持续性乳酸性酸中毒肾上腺素能激活和继发于一般骨骼肌肌病的劳力不耐受作为肥厚型心肌病。最终，心肌增厚和心功能障碍进展至终末期。阶段心力衰竭需要心脏移植。目前尚无任何可用于该患者群体的疾病缓解疗法。然而，腺- 相关病毒（AAV）介导的基因替代疗法已成为治疗疾病的强大策略遗传性单基因疾病的修饰。我们研究的长期目标是开发一种治疗方法治疗 SLC25A4 缺陷的基因替代策略。该提案的目的是进一步描述疾病表型以及合成和评估重组 AAV (rAAV) 载体的功效在患者来源的细胞系和类器官模型的体外模型中。本研究的中心假设建议认为 AAV 介导的基因替换可以改善生化和功能效应发现SLC25A4缺陷，可以更果断地阻止疾病进展。本次活动的具体目标提案是： 1. 表征患者来源的细胞系中 SLC25A4 缺陷表型。 2. 合成重组 AAV 载体，将密码子优化的 SLC25A4 cDNA 递送至骨骼肌细胞和心肌细胞。 3. 评估 AAV-SLC25A4 病毒转导在患者来源的细胞系中的功效。这些实验将增进我们对 SLC25A4 分子机制的理解缺陷，并让我们能够评估 AAV 平台在相关临床前模型中的功效。此外，我在这次研究期间获得的技能将有助于我成为一名独立调查员以及一位专注于转化基因替代疗法开发的外科医生科学家。