Translational studies of GAA deficiency in bioengineered human muscle

生物工程人体肌肉中 GAA 缺乏的转化研究

基本信息

批准号：
8737169
负责人：
Nenad Bursac
金额：
$ 33.13万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-18 至 2018-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8737169
关键词：
3-Dimensional Acids Adjuvant Adjuvant Chemotherapy Adjuvant Therapy Adrenergic Agonists Alpha-glucosidase Alternative Therapies Autophagocytosis Biochemical Biomedical Engineering Biopsy Body Size Candidate Disease Gene Cardiac Cardiac Myocytes Cell Culture Techniques Cells Child Chronic Clinical Clinical Treatment Clinical Trials Degenerative Disorder Development Disease Disease model Dorsal Dose Electric Stimulation Engineering Enzymes Exercise FDA approved Fiber Follistatin Future Genes Glucan 1,4-alpha-Glucosidase Glycogen Glycogen storage disease type II Heart Heart failure Histology Human Human Engineering Implant In Vitro Individual Intervention Intravenous infusion procedures Knock-out Knockout Mice Lead Life Metabolic Methods Modeling Molecular Monitor Mus Muscle Muscle Cells Muscle Fibers Muscle Weakness Muscle function Muscle satellite cell Myoblasts Myocardium Myopathy Needles Nude Mice Optics Outcome Output Patients Pattern Performance Pharmaceutical Preparations Pharmacotherapy Physiological Physiology Principal Investigator Property Recombinants Respiratory Diaphragm Role Serotyping Severity of illness Skeletal Muscle Striated Muscles System Testing Tetanus Therapeutic Time Tissue Engineering Tissues Treatment Efficacy Validation Vascularization base cell injury clinically relevant disease phenotype dosage drug candidate enzyme deficiency enzyme replacement therapy functional outcomes gene therapy glucosidase human IGF2R protein human tissue in vivo infancy minimally invasive mouse model muscle engineering novel prevent public health relevance receptor expression respiratory response satellite cell screening skeletal therapeutic gene translational study vector

项目摘要

DESCRIPTION (provided by applicant): Glycogen storage disease type II (Pompe disease) is a fatal degenerative disease caused by the deficiency of acid-alpha glucosidase (GAA) or acid maltase. This disease is characterized by progressive myopathy resulting from the accumulation of lysosomal glycogen in skeletal and cardiac muscle cells. Enzyme replacement therapy (ERT) with recombinant human GAA is the only FDA-approved treatment for Pompe disease, which despite being beneficial, is highly expensive and inefficient, requiring enzyme doses 100-fold greater than those used for other lysosomal disorders. Furthermore, the ability of ERT to correct important aspects of the disease including autophagy, glycogen accumulation, and low exercise capacity, remains questionable. Therefore, the need for the development of alternative or adjuvant therapies to ERT is obvious, and although the mouse GAA knockout (GAA-KO) model is often utilized for this purpose, the differences in size and physiology of mice and humans and less severe disease phenotype in mice limit the translational utility of these studies. Human cells isolated from patients' muscle biopsies offer an alternative system to study muscle disease in vitro, however, no methods exist to generate functional contractile muscle fibers starting from human muscle cells. In this project we for the first time describe engineering of contractile, electrically responsive human muscle tissues ("bioartificial muscle") made of primary myogenic cells obtained using standard muscle biopsies from normal individuals and Pompe disease patients. We propose to utilize these 3D cell cultures as a predictive in vitro screen for candidat drug and gene therapeutics for human muscle disease. By combining bioengineering and clinical expertise of the two principal investigators, we will carry out a set of translational in itro and in vivo studies in order to screen and validate alternative and adjuvant drug and gene therapies for Pompe disease. In particular, we will: 1) Optimize functional properties of healthy and Pompe disease human bioartifical muscle tissues and systematically characterize their molecular, metabolic and functional properties, 2) Mechanistically study novel candidate drug and AAV therapies for Pompe disease using GAA-KO mice, and 3) Screen the efficacy of these candidate approaches in vitro using engineered human Pompe disease muscle and further validate the most promising therapies in vivo using a novel humanized mouse model of Pompe disease. In the future, the experimental framework established in this project will allow us to undertake similar translational studies to aid treatment of other skeletal and cardiac muscle disorders.

描述（申请人提供）：II型糖原累积病（庞贝病）是一种因酸性-α葡萄糖苷酶（GAA）或酸性麦芽糖酶缺乏而引起的致命退行性疾病。这种疾病的特征是由于溶酶体糖原在骨骼肌和心肌细胞中积累而导致的进行性肌病。使用重组人 GAA 的酶替代疗法 (ERT) 是 FDA 批准的唯一治疗庞贝病的疗法，尽管这种疗法有益，但价格昂贵且效率低下，需要的酶剂量比其他溶酶体疾病大 100 倍。此外，ERT 纠正疾病重要方面（包括自噬、糖原积累和运动能力低下）的能力仍然值得怀疑。因此，开发 ERT 的替代疗法或辅助疗法的必要性是显而易见的，尽管小鼠 GAA 敲除 (GAA-KO) 模型经常用于此目的，但小鼠和人类在体型和生理学上的差异以及不太严重的疾病小鼠的表型限制了这些研究的转化效用。人体细胞从患者的肌肉活检中分离出来提供了一种在体外研究肌肉疾病的替代系统，但是，不存在从人类肌肉细胞开始产生功能性收缩肌纤维的方法。在这个项目中，我们首次描述了由原代肌原细胞制成的可收缩、电响应的人体肌肉组织（“生物人工肌肉”）的工程，这些原代肌原细胞是使用正常个体和庞贝病患者的标准肌肉活检获得的。我们建议利用这些 3D 细胞培养物作为人类肌肉疾病候选药物和基因疗法的体外预测筛选。通过结合两位主要研究人员的生物工程和临床专业知识，我们将开展一系列体外和体内转化研究，以筛选和验证庞贝氏病的替代和辅助药物及基因疗法。具体来说，我们将：1) 优化健康和庞贝氏症人类生物人工肌肉组织的功能特性，并系统地表征其分子、代谢和功能特性，2) 使用 GAA-KO 小鼠从机制上研究庞贝氏症的新候选药物和 AAV 疗法， 3) 使用工程化的人类庞贝氏症肌肉在体外筛选这些候选方法的功效，并使用新型庞贝氏症人源化小鼠模型进一步验证最有前途的体内疗法。未来，该项目建立的实验框架将使我们能够进行类似的转化研究，以帮助治疗其他骨骼和心肌疾病。