Interrogation of retroelement-derived proteins for functional gene transfer

功能性基因转移中逆转录元件衍生蛋白的研究

• 批准号: 10746395
• 负责人: Blake Lash
• 金额: $ 4.36万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10746395
• 关键词: Adenoviruses; Antibodies; Binding; Binding Proteins; Binding Sites; Bioinformatics; Biological; Biological Assay; Biological Process; Capsid; Capsid Proteins; Cell Differentiation process; Cells; Central Nervous System; Clinic; Communication; Computational Biology; Cre lox recombination system; Dependovirus; Development; Disease; Dose; Engineering; Environment; Epitopes; Evolution; Gene Delivery; Gene Transduction Agent; Gene Transfer; Generations; Genes; Genetic; Genetic Diseases; Goals; Homologous Gene; Human Genome; Immune response; Immune system; Immunity; Immunoprecipitation; In Vitro; Inflammation; Innate Immune Response; Investigation; Knock-in; Knock-out; Learning; Lentivirus; Maps; Measures; Mediating; Mentorship; Messenger RNA; Methods; Molecular Biology; Monitor; Mus; Nucleic Acid Binding; Nucleic Acids; Organ; Pathogenicity; Patients; Protein Secretion; Proteins; RNA; RNA Binding; Reporter; Research; Research Personnel; Retroelements; Retrotransposon; Retroviridae; Risk; Role; Serum; Structure; System; T cell response; Techniques; Technology; Tertiary Protein Structure; Therapeutic; Tissues; Tropism; Validation; Viral; Viral Vector; Virus; Work; bioluminescence imaging; body system; candidate identification; cell type; cytokine; delivery vehicle; design; env Gene Products; extracellular vesicles; gene therapy; genome sequencing; human embryonic stem cell line; immunogenic; immunogenicity; in vivo; intercellular communication; interest; mammalian genome; mutant; nanoparticle; nervous system disorder; nucleic acid delivery; programs; response; self assembly; side effect; skills; stem cells; therapeutic gene; tool; vector

Project Summary and Abstract Gene therapy enables the treatment of a large number of genetic diseases through delivery of nucleic acids striking at the root of the disease. This is advantageous because it is highly modular, allowing for a number of different cargo nucleic acids to be delivered depending on the disease cause. As such, the ideal gene therapy delivery vector would be able to carry a variety of cargo, deliver this in a targeted manner, and accommodate a range of cargo sizes. There are a number of techniques utilized to deliver nucleic acids including viral systems like adenovirus, adeno-associated virus (AAV), and lentivirus, as well as non-viral methods including nanoparticles. Although these therapies can be successful, a key limitation to currently used vectors is the immune response which can lead to ineffective delivery of nucleic acid cargo. There is currently a need to develop effective and non-immunogenic delivery vehicles for gene therapy for a wide range of diseases, including neurological disease, for which effective delivery vehicles have yet to be designed. To this end, mammalian genomes contain numerous virus-like genes, some of which have been co-opted by their host cells for important functions. Among these are homologs of gag, which encodes the capsid protein. We hypothesize that endogenous genes encoding a capsid domain have the ability to self-assemble into capsids and mediate intercellular communication by binding, secreting, and delivering nucleic acid cargos. We propose to explore and re-engineer endogenous capsid-containing proteins for use as gene therapy vectors. We hypothesize that delivery vehicles composed entirely of self proteins will be more effective than standard vectors as they could be non-immunogenic. Here we propose to use an approach combining in vitro characterization, re-engineering, and in vitro and in vivo validation to identify candidate proteins and learn how they can be re-engineered. These systems will ideally be modular, having both programmable cargo and tropism to treat a range of diseases. We hope that by identifying and re-engineering these systems, the resulting fully endogenous delivery vehicle will be useful for efficient, reprogrammable, and non-immunogenic gene delivery. With the goal of becoming an independent investigator, this project will also support development of computational biology skills, molecular biology expertise as well as mentorship and scientific communication skills. These will be supported by the excellent research environment at the Broad Institute and MIT.

项目摘要和摘要 基因疗法通过在触及在 疾病的根源。这是有利的，因为它是高度模块化的，允许许多不同的货物核心 取决于疾病的原因，要递送酸。因此，理想的基因疗法递送向量将能够 携带各种货物，以目标方式交付此货物，并容纳各种货物尺寸。有很多 用于输送核酸的技术，包括病毒系统，如腺病毒，腺相关病毒（AAV）和 慢病毒以及包括纳米颗粒在内的非病毒方法。尽管这些疗法可以成功，这是关键 当前使用的载体的局限性是免疫反应，可能导致核酸货物的无效输送。 目前有必要开发有效且非免疫原性递送工具进行基因治疗，以进行广泛的范围 尚未设计有效递送车的神经疾病在内的疾病。 为此，哺乳动物基因组包含许多类似病毒的基因，其中一些是由其宿主选择的 重要功能的细胞。其中包括编码衣壳蛋白的GAG的同源物。我们假设这一点 编码衣壳结构域的内源基因具有自组装成衣壳并介导细胞间的能力 通过结合，分泌和递送核酸兑换来通信。我们建议探索和重新设计 内源性衣壳蛋白作为基因疗法载体。我们假设组成的运输车辆 自我蛋白质的完全比标准载体更有效，因为它们可能是非免疫原性的。我们在这里提出 使用一种结合体外表征，重新设计以及体外和体内验证的方法 候选蛋白质并了解如何重新设计它们。这些系统理想情况下将是模块化的，两者都有 可编程的货物和对流式治疗各种疾病。我们希望通过识别和重新设计这些 系统，由此产生的完全内源性输送车将有效，可重新编程和 非免疫原性基因递送。 为了成为一名独立调查员，该项目还将支持计算的开发 生物学技能，分子生物学专业知识以及指导和科学沟通技巧。这些将是 在广大研究所和麻省理工学院的出色研究环境的支持下。