Bioengineering of phage-derived particles as a discovery platform for muscle gene therapy

噬菌体衍生颗粒的生物工程作为肌肉基因治疗的发现平台

• 批准号: 10758371
• 负责人: Uyanga Tsedev
• 金额: $ 33.46万
• 依托单位: GENSAIC, INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10758371
• 关键词: Address; Affect; Bacteriophage M13; Bacteriophages; Bar Codes; Biodistribution; Biomedical Engineering; Capsid; Capsid Proteins; Cell Nucleus; Cells; Clinic; Clinical; Code; Cytomegalovirus; DNA; Data; Dependovirus; Deposition; Derivation procedure; Development; Directed Molecular Evolution; Duchenne muscular dystrophy; Dystrophin; Engineering; Evolution; Exhibits; Facioscapulohumeral Muscular Dystrophy; Formulation; Future; Gene Delivery; Gene Expression; Genes; Genetic; Genetic Diseases; Genetic Medicine; Goals; Human; Immune; Immune Evasion; Immunity; Immunologic Surveillance; In Vitro; Lead; Libraries; Lipids; Liver; Lung; Mammalian Cell; Messenger RNA; Muscle; Muscle Cells; Muscle Development; Muscle function; Muscular Atrophy; Myoblasts; Myopathy; Myotonic Dystrophy; Nuclear; Output; Patients; Phase; Proteins; RNA; Rare Diseases; Resources; Retina; Small Business Innovation Research Grant; Surface; Technology; Testing; Therapeutic; Tissues; Transfection; Translations; Variant; Viral Genes; Work; cell type; delivery vehicle; design; design and construction; disease-causing mutation; effective therapy; gene therapy; immunogenicity; improved; in vivo; large scale production; manufacturability; manufacture; manufacturing systems; muscle degeneration; muscular dystrophy mouse model; next generation; particle; promoter; protein expression; rare genetic disorder; rational design; screening; targeted treatment; trafficking; transduction efficiency

PROJECT SUMMARY Gene therapy provides a functional cure for patients with genetic disorders and is particularly relevant to the treatment of monogenic rare diseases (those caused by a single gene), such as Duchenne muscular dystrophy (DMD). Adeno-associated virus (AAV) is the most prevalent and clinically successful viral gene therapy. However, as a delivery platform, it exhibits significant limitations when it comes to immunogenicity, cargo ca- pacity, manufacturability, and ease of cell type-specific targeting. Other platforms, such as lipid nanoparti- cle/mRNA formulations, can be manufactured at scale but only induce untargeted and transient gene expres- sion. Therefore, while each has its strengths and weaknesses, there is currently no platform available that brings accessible and durable gene therapy to rare genetic disorders. The goal of M13-Tx is to develop a next-generation in vivo gene delivery platform that addresses the current key limitations of existing gene therapy technologies. This platform is based on phage-derived particles (PDPs) – an immune-privileged, easily engineerable, and efficiently produced derivation of M13 bacteriophage – spe- cifically designed for the transduction of human cells. By the combination of directed evolution and rational de- sign, these PDPs can be engineered to target any tissue and deliver up to 20kb DNA cargo. In this Phase I SBIR, M13-Tx proposes to engineer and screen PDPs to successfully deploy a cargo carrying the functional copy of DMD gene coding for dystrophin protein in muscle tissue in vivo. In contrast to existing AAVs, which can only pack a fraction of the DMD coding sequence, the PDPs will deploy the full DMD gene only in muscle tissue for durable and redoseable treatment of Duchenne muscular dystrophy. The final product of this proposal is lead PDPs, ready to be tested in the DMD mouse model. The lead PDPs and muscle-specific deployment can be utilized across rare diseases caused by mutations of different genes but manifested in the same tissue (e.g., Myotonic Dystrophy and Facioscapulohumeral Muscular Dystrophy). Moreover, the same workflow can be used to find PDPs for other difficult-to-target tissues (e.g., lung and CNS). The proposed study will serve as a proof-of-concept, showing that PDPs can be selected for specific gene delivery purpose, given their engineerable coat and easily modified cargos. Developing this technology will revolutionize gene therapy by presenting a versatile and affordable gene delivery platform.

项目概要 基因疗法为遗传性疾病患者提供功能性治愈，尤其与 治疗单基因罕见疾病（由单一基因引起的疾病），例如杜氏肌营养不良症 （DMD）。腺相关病毒（AAV）是最流行且临床上最成功的病毒基因疗法。 然而，作为一个交付平台，它在免疫原性、货物运输方面表现出很大的局限性。 细胞类型特异性靶向的透明度、可制造性和易用性。其他平台，例如脂质纳米颗粒 cle/mRNA 制剂，可以大规模生产，但只能诱导非靶向和瞬时基因表达 锡安。因此，虽然各有其优点和缺点，但目前还没有可用的平台 为罕见的遗传疾病带来可及且持久的基因疗法。 M13-Tx的目标是开发下一代体内基因传递平台，解决当前的问题 现有基因治疗技术的主要局限性。该平台基于噬菌体衍生颗粒（PDP） – 一种具有免疫特权、易于工程设计且高效生产的 M13 噬菌体衍生品 – 特殊用途 专为人类细胞的转导而设计。通过定向进化和理性设计的结合 迹象表明，这些 PDP 可以被设计为靶向任何组织并输送高达 20kb 的 DNA 货物。 在第一阶段 SBIR 中，M13-Tx 建议设计和筛选 PDP，以成功部署货运 体内肌肉组织中编码肌营养不良蛋白的 DMD 基因的功能副本。与现有的相比 AAV 只能包装 DMD 编码序列的一小部分，PDP 将部署完整的 DMD 基因 仅在肌肉组织中对杜氏肌营养不良症进行持久且可重复给药的治疗。 该提案的最终产品是领先的 PDP，准备在 DMD 小鼠模型中进行测试。领先的 PDP 肌肉特异性部署可用于治疗由不同基因突变引起的罕见疾病 但表现在相同的组织中（例如强直性肌营养不良和面肩肱型肌营养不良）。 此外，相同的工作流程可用于寻找其他难以靶向的组织（例如肺和组织）的 PDP。 中枢神经系统）。拟议的研究将作为概念验证，表明可以针对特定的情况选择 PDP 考虑到其可设计的外壳和易于修改的货物，基因传递的目的。开发这项技术 将通过提供多功能且负担得起的基因传递平台彻底改变基因治疗。