Optimizing membrane repair for the treatment of Duchenne muscular dystrophy

优化膜修复治疗杜氏肌营养不良症

基本信息

批准号：
9910186
负责人：
Noah Weisleder
金额：
$ 22.48万
依托单位：
MYOS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-17 至 2020-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9910186
关键词：
Affect Amino Acid Motifs Amino Acids Animal Model Animal Testing Antibodies Binding Binding Proteins Biological Assay Biological Markers Biotechnology Blood Circulation Cardiac Cardiovascular Diseases Cardiovascular system Cell Death Cell membrane Chronic Clinical Clinical Trials Collaborations Complementary therapies Complex Cultured Cells DNA Development Diffuse Disease Doctor of Philosophy Drug Kinetics Duchenne muscular dystrophy Dystroglycan Dystrophin Engineering Enzyme-Linked Immunosorbent Assay Formulation Future Genes Goals Half-Life Human Intravenous Knockout Mice Laboratories Lead Life Expectancy Link Measures Membrane Membrane Proteins Mus Muscle Muscle Cells Muscle Development Muscle function Muscular Atrophy Muscular Dystrophies Mutation Myocardial Infarction Myocardium Myopathy Necrosis Neurodegenerative Disorders Ohio Outcome Measure Pathology Patient-Focused Outcomes Patients Persons Pharmacologic Substance Phase Phosphatidylserines Polyethylene Glycols Predisposition Preparation Property Protein Engineering Proteins Publishing Recombinant Proteins Recombinants Research Role Serum Site Skeletal Muscle Small Business Technology Transfer Research Source Structural Protein TRIM Motif Technology Tertiary Protein Structure Testing Therapeutic Tissues Toxic effect Universities base boys commercialization congenic deletion analysis improved improved outcome in vivo mdx mouse mouse model muscle form muscle physiology novel novel therapeutics overexpression pharmacokinetic characteristic pre-clinical protein folding protein function protein structure repaired restoration skeletal skeletal preservation standard of care therapeutic development therapeutic protein therapy development three dimensional structure young man

项目摘要

PROJECT ABSTRACT The long-term goal of this project is to optimize a protein therapeutic for Duchenne muscular dystrophy (DMD), and potentially other muscle diseases, that will enhance the repair capacity of muscle cell membranes that are compromised by mutations in the dystrophin/dystroglycan complex. Mutations in the dystrophin/dystroglycan complex result in Duchenne muscular dystrophy. Myos Inc. is developing a novel recombinant construct of the tripartite protein 72/mitsugumin 53 (TRIM72/MG53), an essential regulator of membrane repair in skeletal and cardiac muscle. rhMG53 therapy ameliorates disease pathology in the dystrophin-null mouse model, strongly suggesting that it may enhance repair and restoration of muscle function in DMD. However, its large size and short serum half-life make rhMG53 unsuitable for protein therapy. Therefore, the objective of this Phase I STTR project is to engineer the rhMG53 protein, the result of which will be called MyoTRIM, for use in treating DMD by optimizing its functional and pharmacokinetic (PK) properties. This STTR project is a collaboration with Noah Weisleder, Ph.D. (Ohio State University), who is the PI. Aim 1 is to engineer a compact rhMG53 protein containing key moieties that are required for membrane repair. Deletion analysis and protein engineering approaches will be used to selectively delete regions that are not predicted to affect protein folding. This will reduce protein size from the current 53 kDa to <24 kDa while retaining the essential functional domains and 3D structure. Aim 2 is to improve the PK characteristics of the engineered rhMG53 protein by PEGylation. A panel of three candidate rhMG53 construct resultant from Aim 1 will be modified by covalent and/or non-covalent attachment of polyethylene glycol (PEG) in order to extend its serum half-life from the current 4 hr to >12 hr. Protein function will be tested in an ex vivo membrane repair assay and in the D2.B10 (DBA/2-congenic) Dmdmdx (D2- mdx) mouse model of DMD. Outcome measures will include serum half-life, protein concentrations in serum and target tissues, and serum biomarkers for skeletal and cardiac muscle membrane integrity. This novel, truncated, functional PEGylated rhMG53 construct has great potential to improve muscle membrane repair to treat fatal muscular dystrophies and other forms of muscle disease, independent of gene or mutation class. By enabling membrane resealing and preservation of skeletal and cardiac muscle function, it would provide a complementary treatment approach to other therapeutic efforts now in development. It also may provide a platform technology to target other diseases involving compromised membrane integrity or necrotic cell death, such as cardiovascular disease and neurodegenerative disorders. Successful completion of this Phase I STTR project will result in a novel rhMG53-PEG that is suitable for therapeutic development.

项目摘要该项目的长期目标是优化杜氏肌肉蛋白疗法营养不良（DMD）和潜在的其他肌肉疾病，这将增强肌肉的修复能力肌细胞膜因肌营养不良蛋白/肌营养不良聚糖复合物的突变而受到损害。肌营养不良蛋白/肌营养不良聚糖复合物的突变会导致杜氏肌营养不良症。迈奥斯公司是开发三联蛋白 72/mitsugumin 53 (TRIM72/MG53) 的新型重组结构，骨骼肌和心肌膜修复的重要调节剂。 rhMG53疗法可改善疾病肌营养不良蛋白缺失小鼠模型的病理学，强烈表明它可以增强修复和恢复 DMD 中的肌肉功能。然而，rhMG53 体积大且血清半衰期短，因此不适合用于蛋白质疗法。因此，这一阶段 STTR 项目的目标是改造 rhMG53 蛋白，其结果将被称为 MyoTRIM，通过优化其功能和功能来治疗 DMD 药代动力学 (PK) 特性。该 STTR 项目是与 Noah Weisleder 博士合作的。（俄亥俄州州立大学），他是 PI。目标 1 是设计一种紧凑的 rhMG53 蛋白，其中包含膜修复所需的关键部分。删除分析和蛋白质工程方法将用于选择性删除不存在的区域。预计会影响蛋白质折叠。这会将蛋白质大小从当前的 53 kDa 减少到 <24 kDa，同时保留基本的功能域和 3D 结构。目标 2 是通过聚乙二醇化改善工程化 rhMG53 蛋白的 PK 特性。三人小组由目标 1 产生的候选 rhMG53 构建体将通过共价和/或非共价连接进行修饰聚乙二醇 (PEG)，以将其血清半衰期从目前的 4 小时延长至 >12 小时。蛋白质功能将在离体膜修复测定和 D2.B10（DBA/2-同源）Dmdmdx（D2- mdx) DMD 小鼠模型。结果测量包括血清半衰期、血清中的蛋白质浓度和靶组织以及骨骼和心肌膜完整性的血清生物标志物。这种新颖的、截短的、功能性聚乙二醇化 rhMG53 构建体具有改善肌肉的巨大潜力膜修复可治疗致命性肌营养不良症和其他形式的肌肉疾病，与基因无关或突变类。通过膜重新密封和保存骨骼和心肌功能，将为目前正在开发的其他治疗方法提供补充治疗方法。它还可能提供一种平台技术来针对涉及膜完整性受损的其他疾病或坏死性细胞死亡，例如心血管疾病和神经退行性疾病。顺利完成该一期 STTR 项目的进展将产生一种适合治疗开发的新型 rhMG53-PEG。