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杜氏肌营养不良的蛋白质治疗方法。 营养不良（DMD）和潜在的其他肌肉疾病，这将提高修复能力， 肌细胞膜被肌营养不良蛋白/肌营养不良蛋白聚糖复合物中的突变损害。 肌营养不良蛋白/肌营养不良蛋白聚糖复合物中的突变导致杜氏肌营养不良症。Myos Inc.是 开发了一种新的三联蛋白72/Mitsugumin 53（TRIM 72/MG 53）重组构建体， 骨骼肌和心肌膜修复的重要调节因子。rhMG 53治疗改善疾病 病理学在肌营养不良蛋白无效的小鼠模型，强烈表明它可以增强修复和恢复， 肌肉功能的变化然而，其大的尺寸和短的血清半衰期使得rhMG 53不适合用于 蛋白质疗法因此，该I期STTR项目的目标是工程化rhMG 53蛋白， 其结果将被称为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博士合作的。（俄亥俄州 州立大学），他是PI。 目的1是工程化一个紧凑的rhMG 53蛋白含有膜修复所需的关键部分。 缺失分析和蛋白质工程方法将用于选择性地删除不适合的区域。 会影响蛋白质折叠这将使蛋白质大小从目前的53 kDa减少到<24 kDa， 保留了基本的功能结构域和三维结构。 目的二是通过聚乙二醇修饰来改善rhMG 53工程蛋白的PK特性。三人小组 将通过共价和/或非共价连接来修饰从Aim 1得到的候选rhMG 53构建体 为了将其血清半衰期从目前的4小时延长到>12小时，我们使用了聚乙二醇（PEG）。 功能将在离体膜修复试验和D2.B10（DBA/2-同源）Dmdmdx（D2-同源）中进行测试。 mdx）DMD的小鼠模型。结果测量将包括血清半衰期、血清中的蛋白浓度 和靶组织，以及骨骼肌和心肌膜完整性的血清生物标志物。 这种新型的截短的功能性聚乙二醇化rhMG 53构建体具有改善肌肉的巨大潜力， 膜修复治疗致命性肌营养不良症和其他形式的肌肉疾病，不依赖于基因 或突变类。通过使膜重新密封和保存骨骼肌和心肌功能， 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 可以提供一种平台技术，以靶向涉及受损膜完整性的其他疾病， 坏死性细胞死亡，如心血管疾病和神经退行性疾病。成功完成 该I期STTR项目的研究将产生适合于治疗开发的新型rhMG 53-PEG。