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.

项目总结 基因治疗为遗传性疾病患者提供了一种功能性治疗，尤其与 治疗单基因罕见疾病(由单基因引起的疾病)，如杜氏肌营养不良 (DMD)。腺相关病毒(AAV)是目前应用最广泛、临床上最成功的病毒基因治疗方法。 然而，作为一个交付平台，它在免疫原性方面表现出很大的限制，货物可以- 灵活性、可制造性和针对特定细胞类型的目标定位的简便性。其他平台，如脂质纳米颗粒- 可以规模化生产，但只能诱导非靶向和瞬时的基因表达。 西翁。因此，虽然每个平台都有其优势和劣势，但目前还没有可用的平台 为罕见的遗传疾病带来可获得和持久的基因治疗。 M13-TX的目标是开发下一代体内基因传递平台，以解决当前 现有基因治疗技术的关键限制。该平台基于噬菌体衍生颗粒(PDP) -一种免疫特权、易于工程和高效生产的M13噬菌体衍生体- 专为人类细胞的转导而设计。通过定向进化和理性去中心化的结合 签名，这些PDP可以针对任何组织进行工程设计，并提供高达20kb的DNA货物。 在此第一阶段SBIR中，M13-TX建议设计和筛选PDP，以成功部署货物运输 肌肉组织中肌营养不良蛋白DMD基因的功能拷贝。与现有的 AAVs只能包装DMD编码序列的一小部分，PDP将部署完整的DMD基因 仅在肌肉组织中使用，可持久和可修复地治疗杜氏肌营养不良症。 这项提议的最终产品是铅PDP，准备在DMD小鼠模型中进行测试。主要的PPP 肌肉特异性部署可以用于治疗由不同基因突变引起的罕见疾病 但表现在同一组织中(例如，强直性肌营养不良和面肩肩周肌营养不良)。 此外，可以使用相同的工作流来为其他难以靶向的组织(例如，肺和 CNS)。拟议的研究将作为概念验证，表明PPP可以被选择用于特定的 基因传递目的，考虑到它们可工程的皮毛和容易修改的货物。开发这项技术 将通过提供一个多功能和负担得起的基因交付平台来彻底改变基因治疗。