Continuous Evolution of Proteins with Novel Therapeutic Potential

具有新治疗潜力的蛋白质的不断进化

基本信息

批准号：
10588186
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
项目类别：
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/10588186
关键词：
Age Agriculture Alleles Animal Model Antibodies Bacteriophages Binding Proteins Biological Process Biological Sciences Biotechnology Brain Diseases Cell model Cells Clinical Clustered Regularly Interspaced Short Palindromic Repeats DNA DNA-Directed RNA Polymerase Development Disease Enzymes Escherichia coli Evolution Foundations Genes Genetic Diseases Genome Goals Guide RNA Human Human Genetics Infection Insecticides Intervention Laboratories Malignant neoplasm of brain Mediating Methods Modality Mutagenesis 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 editor clinically relevant disease-causing mutation drug-sensitive functional genomics gene product genetic manipulation genome editing genomic tools human disease in vivo innovation interest macromolecule mutation correction new technology next generation novel novel therapeutics nuclease prime editing prime editor programs protein E protein degradation research study small molecule success therapeutic development therapeutic target tool

项目摘要

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：具有新治疗潜力的蛋白质的持续进化体内基因和基因产物的直接操作具有巨大的治疗潜力，并且许多实现这些目标的策略正在迅速推进到临床应用，可以操纵DNA，活细胞中的 RNA 和蛋白质，包括能够精确校正基因组编辑技术体内引起疾病的突变，已经证明了这种方法对于研究和应用的前景。虽然许多这些方法在初步研究中显示出希望，蛋白质通常需要广泛的开发和定制才能获得所需的活性、特异性和稳定性作为有影响力的研究工具或治疗开发的先导。模式、快速生成蛋白质的方法继续以惊人的速度发展理想情况下，需要具有定制功能的方法，并且可以应用于许多领域。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传统的实验室进化方法更快，所需的研究人员最少我们已经证明了 PACE 通过新的蛋白质进化出许多不同类别的蛋白质的能力。以及改变的活性、特异性和其他所需特性，例如大肠杆菌中的可溶性表达。使用 PACE 进化而来的基因组在多种非细菌环境中显示出广泛的编辑效用，包括基因组试剂已应用于挽救遗传性疾病的人类细胞和动物模型以及杀虫蛋白这些发展使 PACE 成为一种广泛适用且高度可行的技术。产生治疗和生物技术相关蛋白质的技术。我们建议应用 PACE 来进化具有治疗潜力的新型蛋白质，或者使新的蛋白质成为可能这些蛋白质包括下一代精确基因组编辑技术。可以更容易地在体内递送或更有效且具有临床相关性的药物；裂解与神经退行性疾病和脑癌有关的内源性蛋白质靶点的蛋白酶；以及能够实现药物诱导的靶蛋白降解的小分子结合蛋白。为纠正导致人类基因突变的创新治疗策略奠定基础此外，还可以将自传递蛋白酶重新编程为治疗脑部疾病的催化药物。创建药物敏感的等位基因，使感兴趣的蛋白质以小分子依赖性方式降解以这种方式，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总的来说，这项研究整合了强大的蛋白质。利用酶精确操纵基因组和蛋白质组以促进进步的进化技术治疗学科学。