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

• 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
• 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

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.

项目摘要/摘要 线粒体氧化磷酸化（OXPHOS）的破坏与发展 生化改变通常会影响高能量需求的组织，尤其是骨骼和心脏 肌肉。遗传的常染色体隐性骨骼肌病和肥厚性心肌病已连接 由于溶质中的翻新突变，核DNA编码的线粒体蛋白的功能丧失 载体家族25，成员4（SLC25A4； C.523delc，P.Q175RFXX38）。 SLC25A4编码心肌 腺嘌呤核苷酸易位剂1（ANT1，SLC25A4）的同工型，在野生型状态下是关键的 线粒体代谢的成分。 SLC25A4缺乏症患者表现出乳酸性酸中毒，持续 肾上腺素能激活和继发于一般骨骼肌肉肌病的劳累不耐受性 作为肥厚的心肌病。最终，心肌增厚和心脏功能障碍发展为结束 阶段心力衰竭需要心脏移植。 目前，该患者队列尚无任何修改疾病改良疗法。但是，adeno- 相关病毒（AAV）介导的基因置换疗法已成为疾病的有力策略 遗传性疾病的修饰。我们研究的长期目标是开发治疗性 治疗SLC25A4缺乏症的基因替代策略。该提议的目的是进一步表征 疾病表型以及综合和评估重组AAV（RAAV）载体的功效 在患者衍生的细胞系和类器官模型的体外模型中。中心假设 建议是AAV介导的基因置换可以改善生化和功能效应 SLC25A4缺乏症，可以更具决定性预防疾病进展。这个特定的目的 提案是： 1。表征患者衍生的细胞系中SLC25A4缺乏表型。 2。合成重组AAV载体，用于将密码子优化的SLC25A4 cDNA传递到 骨骼和心肌细胞。 3。评估AAV-SLC25A4病毒转导在患者衍生细胞系中的功效。 这些实验将提高我们对SLC25A4下分子机制的理解 不足以及使我们能够在相关的临床前模型中评估AAV平台的功效。 此外，我在这项奖学金期间将获得的技能将有助于确立我的独立调查员 和外科医生科学家致力于转化基因替代疗法的发展。