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

• 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
• 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

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Duchenne肌肉的蛋白质治疗方法 营养不良（DMD）和可能其他肌肉疾病，这将增强修复能力 肌肉细胞膜因肌营养不良蛋白/营养不良的复合物中的突变而损害。 肌营养不良蛋白/营养不良的复合物中的突变导致Duchenne肌肉营养不良。 Myos Inc.是 开发三方蛋白72/mitsugumin 53（Trim72/mg53）的新型重组构建体，一个 骨骼和心肌修复膜修复的基本调节剂。 RHMG53治疗可以改善疾病 肌营养不良蛋白无小鼠模型中的病理，强烈表明它可以增强修复和恢复 DMD中的肌肉功能。但是，它的大尺寸和短血清半衰期使RHMG53不适合 蛋白质治疗。因此，该阶段ISTTR项目的目的是设计RHMG53蛋白， 其结果将称为Myotrim，用于通过优化其功能和 药代动力学（PK）特性。这个STTR项目是与Noah Weisleder博士的合作。 （俄亥俄州 州立大学），是PI。 AIM 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是 预计会影响蛋白质折叠。这将使蛋白质大小从当前的53 kDa降低到<24 kDa，而 保留基本功能域和3D结构。 AIM 2是通过Pegylation改善工程RHMG53蛋白的PK特性。三个小组 AIM 1产生的候选RHMG53构建体将通过共价和/或非共价附件来修改 聚乙烯乙二醇（PEG）的血清半衰期从当前的4小时延伸到> 12小时。蛋白质 功能将在离体膜修复测定法和D2.b10（DBA/2-综合）DMDMDX中进行测试（D2- DMD的MDX）模型。结果指标将包括血清半衰期，血清中的蛋白质浓度 以及靶组织，以及用于骨骼和心脏肌肉完整性的血清生物标志物。 这种小说，截断，功能性的pegyped RHMG53构建体具有改善肌肉的巨大潜力 膜修复以治疗致命的肌肉营养不良和其他形式的肌肉疾病，与基因无关 或突变类。通过使膜重新密封并保存骨骼和心肌功能， 现在将为现在开发的其他治疗努力提供互补的治疗方法。也是如此 可能会提供平台技术来针对其他涉及膜完整性损害的疾病或 坏死细胞死亡，例如心血管疾病和神经退行性疾病。成功完成 在此I阶段，ISTTR项目将导致适合治疗性发育的新型RHMG53-PEG。