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

基本信息

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
项目类别：
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

项目摘要

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疗法通过递送核酸来攻击基因，从而能够治疗大量遗传疾病。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根据疾病原因输送酸。因此，理想的基因治疗递送载体将能够运载各种货物，有针对性地交付，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。尽管这些疗法可能会成功，但一个关键目前使用的载体的限制是免疫反应，这可能导致核酸货物的无效递送。目前需要开发有效且非免疫原性的递送载体，用于多种基因治疗疾病，包括神经系统疾病，尚未设计出有效的递送载体。为此，哺乳动物基因组包含许多类似病毒的基因，其中一些已被宿主选择细胞发挥重要功能。其中包括编码衣壳蛋白的gag的同源物。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讯。我们建议探索和重新设计用作基因治疗载体的内源性衣壳蛋白。我们假设运载工具由完全由自身蛋白组成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结合体外表征、重新设计以及体外和体内验证的方法来识别候选蛋白质并了解如何重新设计它们。这些系统最好是模块化的，具有可编程的货物和趋向性可治疗一系列疾病。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运载工具将可用于高效、可重新编程和非免疫原性基因传递。为了成为一名独立研究者，该项目还将支持计算技术的发展生物学技能、分子生物学专业知识以及指导和科学沟通技巧。这些将是得到了布罗德研究所和麻省理工学院优良研究环境的支持。