Improving delivery of therapeutic material to skeletal muscle

改善治疗材料向骨骼肌的输送

• 批准号: 9906360
• 负责人: Douglas Paul Millay
• 金额: $ 39.75万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-20 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9906360
• 关键词: Animal Model; Area; Attention; Binding; Cell fusion; Cell surface; Cells; Cellular Membrane; Characteristics; Data; Dependovirus; Development; Dystrophin; Engineering; Event; Fibroblasts; Genes; Genetic; Giant Cells; Goals; Human; Immune system; In Vitro; Investigation; Knowledge; Length; Lentivirus Vector; Location; Membrane; Membrane Proteins; Methods; Modality; Molecular; Muscle; Muscle Cells; Muscle Fibers; Muscle function; Muscle satellite cell; Muscular Dystrophies; Musculoskeletal; Musculoskeletal Diseases; Mutate; Myoblasts; Myopathy; Natural regeneration; Non-Viral Vector; Outcome; Peptides; Phase; Phenotype; Process; Protein Region; Proteins; Regenerative Medicine; Skeletal Muscle; Subfamily lentivirinae; Surface; System; Technology; Testing; Therapeutic; Time; Tissues; Transplantation; Viral; Virus; Work; base; clinically relevant; design; efficacy testing; exosome; functional group; gene correction; gene therapy; improved; in vivo; innovation; mdx mouse; mouse model; nanoparticle; new technology; novel; particle; progenitor; protein function; reconstitution; repaired; transduction efficiency; treatment strategy; vector; virus envelope

Project Summary/Abstract Despite advances in gene therapy, delivery of therapeutic material to a specific tissue remains a challenge. This proposal tackles the long-standing delivery issue in the gene therapy field by engineering novel vehicles for muscle tissue. Our approach to this challenge is to harness the activities of the proteins that directly control myoblast fusion, a process essential for multinucleated skeletal muscle fibers to develop, repair, and regenerate. Fusion of a myoblast membrane with a myofiber membrane allows entry of the progenitor into the syncytium, which is a process that, if understood molecularly and properly engineered, could empower delivery vehicles to transduce muscle. We will leverage our discoveries of Myomaker and Myomerger, which represent the minimal and essential machinery for myoblast fusion, to engineer enveloped viruses and exosomes into specific and efficient vehicles that deliver therapeutic material to muscle cells. Evidence for the challenges associated with muscle gene therapy is the lack of a treatment for genetic muscle diseases such as muscular dystrophy. Adeno-associated virus (AAV) is the current standard for efficacious skeletal muscle gene therapy, but the field has had to reconcile an apparent inability to target muscle stem cells, a goal that would likely need to be achieved for sustained corrective outcomes. The principal rationale of this proposal is that with the discovery of muscle-specific fusogens, it is time to re-examine the potential of non-AAV vectors such as lentiviral vectors and non-viral particles (exosomes) as delivery vehicles that could be used with AAV or independently. Because Myomaker and Myomerger function at the cell surface of myoblasts to drive the membrane remodeling processes necessary for fusion, we propose that their presence on viral envelopes and exosomes will increase entry into muscle. In the R61 phase of this project, we will engineer and optimize Myomaker and Myomerger, or regions of these proteins, on the envelope of viruses and the surface of exosomes. We will also assess and optimize the ability of these engineered vehicles to drive entry into muscle and non-muscle tissues in vitro and in vivo. In the R33 phase, we will validate the use of the delivery vehicles optimized in the R61 phase to test their ability to deliver clinically relevant levels of therapeutic material. Specifically, we will determine if our optimized delivery vehicles can restore dystrophin and improve muscle function in the dystrophin-deficient mdx mouse model. Overall, this work promises to open up a new area of investigation into regenerative medicine by innovating novel delivery vehicles that could be utilized for a myriad of musculoskeletal conditions.

项目摘要/摘要 尽管基因治疗取得了进步，但将治疗材料递送到特定的组织仍然是一个挑战。 该提案通过工程新型车辆解决了基因治疗领域的长期交付问题 用于肌肉组织。我们应对这一挑战的方法是利用直接控制的蛋白质的活动 肌细胞融合，这是多核骨骼肌纤维开发，修复和 再生。肌细胞膜与肌纤维膜的融合允许祖细胞进入 合成剂，这是一个过程，如果可以理解分子和正确设计，则可以增强交付能力 车辆转导肌肉。我们将利用代表Myomaker和Myomerger的发现 肌细胞融合的最小和必需的机械，以将病毒和外泌体笼罩 为肌肉细胞提供治疗材料的特定和高效的车辆。挑战的证据 与肌肉基因治疗相关的是缺乏治疗遗传肌肉疾病（例如肌肉） 营养不良。腺相关病毒（AAV）是当前有效骨骼肌基因治疗的标准， 但是该领域不得不调和明显无法靶向肌肉干细胞的目标，这个目标可能需要 可以实现持续的纠正效果。该提议的主要理由是 发现肌肉特异性的融合原，是时候重新检查非AAV载体的潜力 慢病毒载体和非病毒颗粒（外泌体）作为可以与AAV一起使用或 独立。因为Myomaker和Myomerger在成肌细胞的细胞表面功能以驱动 融合所需的膜重塑过程，我们建议它们在病毒信封上的存在和 外泌体将增加进入肌肉的进入。在该项目的R61阶段，我们将设计和优化 在病毒的信封和表面上 外泌体。我们还将评估和优化这些工程车辆推动进入肌肉的能力 体外和体内的非肌肉组织。在R33阶段，我们将验证使用交付车的使用 在R61阶段进行了优化，以测试其提供临床相关治疗材料水平的能力。 具体来说，我们将确定我们优化的送货车是否可以恢复肌营养不良蛋白并改善肌肉 肌营养不良蛋白缺陷的MDX小鼠模型中的功能。总体而言，这项工作有望为 通过创新可用于无数的新型送货工具来调查再生医学 肌肉骨骼条件。