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：
9076806
负责人：
Nenad Bursac
金额：
$ 10万
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/9076806
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 Health 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 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（由申请人提供）：II型糖原储存疾病（蓬松疾病）是由酸 - α-α葡萄糖酶（GAA）或酸性麦芽瘤的缺乏引起的致命退行性疾病。该疾病的特征是骨骼肌和心肌细胞中溶酶体糖原的积累产生的进行性肌病。重组人GAA的酶替代疗法（ERT）是唯一经FDA批准的庞贝疾病的治疗方法，尽管是有益的，但它非常昂贵且效率低下，需要酶剂量比其他溶酶体疾病使用的酶大100倍。此外，ERT纠正疾病重要方面的能力包括自噬，糖原积累和低运动能力，仍然值得怀疑。因此，对于ERT开发替代或辅助疗法的需求是显而易见的，尽管小鼠和人类的大小和生理学的差异通常用于此目的，但小鼠的大小和生理差异以及小鼠的疾病表型的差异限制了这些研究的翻译实用性。人类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，没有任何方法可以从人类肌肉细胞中产生功能性收缩的肌肉纤维。在这个项目中，我们首次描述了收缩的，电作用的人类肌肉组织（“生物人工肌肉”），该肌肉是由使用正常个体和庞贝病患者的标准肌肉活检获得的原代肌原性细胞制成的。我们建议利用这些3D细胞培养物作为人类肌肉疾病的念珠菌药物和基因疗法的预测性筛查。通过结合两位主要研究人员的生物工程和临床专业知识，我们将进行一系列ITRO和体内研究的翻译，以筛查和验证庞贝疾病的替代性和辅助药物和基因疗法。特别是：1）优化健康和蓬松疾病的人类生物体育肌肉组织的功能性能，并系统地表征其分子，代谢和功能特性，2）机械师使用GAA-KO小鼠的庞贝疾病进行机械研究，并使用这些候选疾病的疾病和3）筛选群体的疗效，并筛选出群体的效率，并筛选出群体的效率，并筛选出群体的效率，并取得了繁琐的效果。使用一种新型的庞贝疾病的人源性小鼠模型在体内疗法。将来，该项目中建立的实验框架将使我们能够进行类似的翻译研究，以帮助治疗其他骨骼和心脏肌肉障碍。