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: 10537044
• 负责人: Blake Lash
• 金额: $ 4.27万
• 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/10537044
• 关键词: Adenoviruses; Antibodies; Binding; Binding Proteins; Binding Sites; Bioinformatics; Biological; Biological Assay; Biological Process; Capsid; Capsid Proteins; Cell Differentiation process; Cells; 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; Human Genome; Immune response; Immune system; Immunity; Immunoprecipitation; In Vitro; Inflammation; Innate Immune Response; Institutes; Investigation; Knock-in; Knock-out; Lead; Learning; Lentivirus; Maps; Measures; Mediating; Mentorship; Messenger RNA; Methods; Molecular Biology; Monitor; Mus; Neuraxis; Nucleic Acid Binding; Nucleic Acids; Organ; Pathogenicity; Patients; Plant Roots; 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; base; bioluminescence imaging; body system; 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; 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.

项目概要和摘要 基因治疗使得能够通过递送核酸来治疗大量的遗传疾病，所述核酸攻击靶点。 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酸被运输。 根据疾病的原因提供酸。因此，理想的基因治疗递送载体将能够 运载各种货物，以有针对性的方式运送货物，并适应各种货物尺寸。有许多 用于递送核酸的技术包括病毒系统，如腺病毒、腺相关病毒（AAV）和腺病毒载体。 慢病毒以及包括纳米颗粒的非病毒方法。虽然这些疗法可以成功，但关键是 对目前使用的载体的限制是免疫应答，其可导致核酸货物的无效递送。 目前需要开发用于基因治疗的有效且非免疫原性的递送载体，用于广泛的免疫缺陷。 这些疾病包括神经系统疾病，尚未设计有效的运载工具。 为此，哺乳动物的基因组中含有大量的病毒样基因，其中一些基因已被其宿主所吸收 细胞的重要功能。其中有gag的同源物，它编码衣壳蛋白。我们假设 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相互作用的能力。 通过结合、分泌和递送核酸货物进行通信。我们建议探索和重新设计 含有内源性衣壳蛋白的蛋白质用作基因治疗载体。我们假设运载工具由 完全的自身蛋白质将比标准载体更有效，因为它们可能是非免疫原性的。在这里我们建议 使用体外表征、再工程以及体外和体内验证相结合的方法， 候选蛋白质，并学习如何重新设计它们。这些系统将是理想的模块化， 可编程货物和向性来治疗一系列疾病。我们希望通过识别和重新设计这些 系统，所得的完全内源性递送载体将可用于高效的、可重新编程的，和 非免疫原性基因递送。 随着成为一个独立的研究者的目标，这个项目也将支持计算的发展， 生物学技能，分子生物学专业知识以及指导和科学沟通技能。这些将是 在布罗德研究所和麻省理工学院的优秀研究环境的支持下。