Continuous Evolution of Proteins with Novel Therapeutic Potential

具有新治疗潜力的蛋白质的不断进化

• 批准号: 10181559
• 负责人: DAVID R LIU
• 金额: $ 62.19万
• 依托单位: BROAD INSTITUTE, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10181559
• 关键词: Age; Agriculture; Alleles; Animal Model; Antibodies; Bacteriophages; Binding Proteins; Biological Process; Biological Sciences; Biotechnology; Brain Diseases; Cell model; Cells; Cleaved cell; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; DNA; Development; Disease; Enzymes; Escherichia coli; Evolution; Foundations; Genes; Genetic Diseases; Genetic Models; Genome; Goals; Guide RNA; Human; Human Genetics; Infection; Insecticides; Intervention; Laboratories; Malignant neoplasm of brain; Mediating; Methods; Modality; Mutagenesis; Mutation; Neurodegenerative Disorders; Peptide Hydrolases; Pharmaceutical Preparations; Plants; Plasmids; Property; Proteins; Proteome; RNA; Research; Research Personnel; Science; Specificity; System; Technology; Therapeutic; Therapeutic Agents; Time; Variant; base; clinically relevant; delta protein; disease-causing mutation; drug-sensitive; functional genomics; gene product; genetic manipulation; genome editing; genomic tools; human disease; in vivo; innovation; interest; macromolecule; new technology; next generation; novel; novel therapeutics; nuclease; protein E; protein degradation; research study; small molecule; success; therapeutic development; therapeutic target; tool

Project Summary: Continuous Evolution of Proteins with Novel Therapeutic Potential The direct manipulation of genes and gene products in vivo has enormous therapeutic potential, and many strategies to achieve these goals are swiftly advancing toward clinical use. Proteins that can manipulate DNA, RNA, and proteins in living cells, including genome editing technologies that enable the precise correction of disease-causing mutations in vivo, have exemplified the promise of such approaches both for research and therapeutic applications. While many of these approaches have shown promise in initial research studies, proteins often require extensive development and tailoring to acquire the activity, specificity, and stability needed to serve as impactful research tools or leads for therapeutic development. As new macromolecular therapeutic modalities continue to be developed at a remarkable rate, methods to generate proteins on a rapid time scale with tailor-made functions are needed. Ideally such methods will be versatile and can be applied to many classes of problems in the life sciences. Our lab developed phage-assisted continuous evolution (PACE), a technology to evolve biomolecules ≥100- fold faster than using conventional laboratory evolution approaches, with minimal required researcher intervention. We have demonstrated the ability of PACE to evolve many different classes of proteins with new and altered activities, specificities, and other desirable properties such as soluble expression in E. coli. Proteins evolved using PACE have shown broad utility in multiple non-bacterial settings, including genome editing agents that have been applied to rescue human cell and animal models of genetic diseases, and insecticidal proteins that kill agricultural pests. These developments establish PACE as a broadly applicable and highly enabling technology for generating therapeutically and biotechnologically relevant proteins. We propose to apply PACE to evolve novel proteins with therapeutic potential, or that enable new technologies for therapeutics discovery. These proteins include next-generation precision genome editing agents that can be more easily delivered in vivo or are more efficient and clinically relevant; self-delivering proteases that cleave endogenous protein targets implicated in neurodegenerative disorders and brain cancer; and small molecule-binding proteins that enable drug-induced target protein degradation. Success would provide a foundation for innovative therapeutic strategies to correct mutations that cause human genetic diseases, and to reprogram self-delivering proteases as catalytic drugs to treat brain diseases. In addition, by creating drug-sensitive alleles that allow a protein of interest to be degraded in a small molecule-dependent manner, the proposed research would establish powerful new functional genomics tools to reveal biological functions and validate therapeutics targets. Collectively, the proposed research integrates powerful protein evolution technologies with enzymes that precisely manipulate genomes and proteomes to advance therapeutics science.

项目摘要：具有新型治疗潜力的蛋白质的连续演变 在体内对基因和基因产物的直接操纵具有巨大的治疗潜力，许多 实现这些目标的策略正在迅速发展临床使用。可以操纵DNA的蛋白质， RNA和活细胞中的蛋白质，包括基因组编辑技术，使得能够精确校正 在体内引起疾病的突变已经举例说明了研究和研究的希望 治疗应用。尽管这些方法中有许多在最初的研究中表现出了希望，但 蛋白质通常需要广泛的开发和裁缝以获取所需的活动，特异性和稳定性 充当有影响力的研究工具或用于治疗开发的潜在客户。作为新的大分子疗法 继续以显着的速度开发方式，即快速产生蛋白质的方法 需要量身定制的功能。理想情况下，此类方法将是多功能的，可以应用于许多 生命科学中的问题类别。 我们的实验室开发了噬菌体辅助连续进化（PACE），这是一种进化生物分子≥100-的技术 折叠速度比使用常规实验室进化方法更快，而所需的研究人员最少 干涉。我们已经证明了速度能够随着新的方式发展许多不同类别的蛋白质 以及改变的活动，特异性和其他期望的特性，例如大肠杆菌中的固体表达。蛋白质 使用速度进化的已在多种非细菌环境中显示出广泛的效用，包括基因组编辑剂 已用于挽救遗传疾病的人类细胞和动物模型和杀虫蛋白 杀死农业害虫。这些事态发展确立了一个广泛适用且高度有利的步伐 用于产生热和生物技术相关蛋白质的技术。 我们建议采用节奏以具有治疗潜力的新蛋白质发展，或者使新的蛋白质启用 治疗发现技术。这些蛋白质包括下一代精度基因组编辑 可以更容易在体内传递或更有效且在临床上相关的代理；自我交付 清除神经退行性疾病和脑癌实施的内源性蛋白质靶标的蛋白酶； 和小分子结合蛋白，可实现药物诱导的靶蛋白降解。成功会 为创新的治疗策略提供了基础，以纠正引起人类遗传的突变 疾病，并将自我删除的蛋白酶作为催化药物进行重新编程，以治疗脑部疾病。另外，由 创建对药物敏感的等位基因，使感兴趣的蛋白质依赖于小分子 拟议的研究将建立强大的新功能基因组学工具来揭示生物学 功能和验证治疗靶标。拟议的研究集体整合了强大的蛋白质 具有精确操纵基因组和蛋白质组的酶的进化技术 治疗科学。