Engineering Smart Antibody-like Protein Scaffolds with precision switches

具有精密开关的工程智能类抗体蛋白支架

基本信息

批准号：
10708167
负责人：
Yubin Zhou
金额：
$ 33.52万
依托单位：
TEXAS A&M UNIVERSITY HEALTH SCIENCE CTR
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：
2022-09-22 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10708167
关键词：
Acute Address Alzheimer&apos s disease model Amyloid beta-42 Animals Antibodies Antigens Antiviral Agents Beverages Biological Biological Assay Biological Models Biological Process Biomedical Research Biophysics Caffeine Cell physiology Cells Cellular biology Chemicals Complement Consumption Cues Data Development Drosophila genus Engineering FDA approved Future Generations Genetic Genome engineering Goals Hepatitis C virus Heterodimerization Huntington Disease Huntington gene Immunotherapy Invaded Kinetics Knock-in Knowledge Light Malignant Neoplasms Methods Mission Modality Modeling Molecular Molecular Immunology Morbidity - disease rate Names Nerve Degeneration Neurodegenerative Disorders Organism Pathway interactions Penetration Peptide Hydrolases Pharmaceutical Preparations Photons Precision therapeutics Process Property Protein Engineering Proteins Public Health Publications Research Resolution Resources Rodent Rodent Model Scaffolding Protein Scheme System Technology Testing Therapeutic Time Tissues Transducers Tumor Immunity United States National Institutes of Health Viral Visualization antibody engineering antibody mimetics biological systems case-by-case basis chimeric antigen receptor T cells clinically relevant cofactor design genetic approach high throughput screening human disease human model improved in vivo in vivo Model innovation interest molecular imaging mortality mutant nano nanobodies nanoparticle neuroregulation neurotoxic novel novel strategies optogenetics photoactivation protein protein interaction public health relevance remote control research and development structural biology synthetic biology theranostics timeline tool translational applications translational barrier tumor immunology wireless

项目摘要

PROJECT SUMMARY / ABSTRACT The goal of this Focused Technology R&D proposal is to develop and apply modular and generalizable engineering approaches to generate smart antibody-like protein scaffolds (APSs) equipped with precision switches, which can be controlled by light or drugs to confer remote control over endogenous proteins and cellular physiology in multiple biological systems. Over the past decade, a variety of chemogenetic and optogenetic tools have been designed to visualize, delocalize, modify, and degrade proteins of interest (POIs). These engineering efforts, nonetheless, often require extensive prior knowledge on the targeted POIs. To regulate endogenous POIs in living cells or organisms, one has to tag POIs with light- or chemical-sensitive modules via genetic knock-in or genome engineering, thereby making the process rather time- and resource- consuming. Furthermore, some of the existing chemo/optogenetic tools still suffer from relatively slow activation kinetics, partial irreversibility, limited choices of chemoswitches , and narrow dynamic ranges of cue- induced changes. To address these challenges, the transdisciplinary team proposes to engineer modular precision switches into single-domain antibody-like protein scaffolds, rather than the endogenous target itself, to confer tight control over POIs and the associated biological activities or pathways. Specially, the team will combine seven selected APSs templates (including nanobody, monobody and affibody) with innovative optogenetic and chemogenetic approaches to develop new generations of light- or chemical-controllable APSs (named as LiAPSs and ChiAPSs, respectively). The prioritized choices of switches include: (i) photons emitting in the blue, far-red, and near infrared (NIR) range (400-800 nm) to diversify the existing repertoire of LiAPSs and significantly improve their kinetic and dynamic properties (Specific Aim 1); (ii) FDA-approved drugs (antivirals) and beverages (caffeine and its metabolites) that promise to reduce barriers for translational applications (Specific Aim 2). These switchable APSs will allow the team to remotely control antibody-antigen recognition and to manipulate endogenous targets in a reversible manner at high temporal and/or spatial resolution. In parallel, the team will demonstrate the applications of engineered smart APSs for acute and precise initiation and termination of biological processes in cellulo, as well as remote in vivo immuno- or neuromodulation in both rodent and Drosophila models of human diseases. Compelling preliminary data have been provided to demonstrate the high feasibility of the proposed new approaches, as well as the team’s mastery of the repertoire of methods, assays, and models described in the application. The innovative molecular toolkit to be generated from the project will offer a wide choices of precision switches to enable many future biological questions and impose a high and sustainable impact to the biomedical field.

项目摘要 /摘要这项重点技术研发建议的目的是开发和应用模块化和可推广生成具有精度的智能抗体样蛋白支架（APS）的工程方法开关，可以通过光或药物来控制，以会议对内源性蛋白质的遥控控制和多种生物系统中的细胞生理。在过去的十年中，各种化学发生和光遗传学工具旨在可视化，分离，修改和降解感兴趣的蛋白质（POI）。尽管如此，这些工程努力通常需要有关目标POI的广泛的先验知识。到调节活细胞或生物中的内源性毒药，必须用光敏感或化学敏感通过遗传敲诈或基因组工程进行模块，从而使过程相当时间和资源 - 消费。此外，一些现有的化学/光遗传学工具仍然相对较慢激活动力学，部分不可逆性，有限的化学开关选择和提示的狭窄动态范围诱导的变化。为了应对这些挑战，跨学科团队提出了工程师模块化的建议精密切换为单域抗体样蛋白支架，而不是内源性靶标本身，对POIS和相关的生物活动或途径进行严格控制。特别是，团队将将七个选定的APS模板（包括纳米机构，单枪和杂物）与创新相结合发育新一代的光或化学可控制的APS的光遗传学和化学遗传学方法（分别为LIAPSS和CHIAPSS）。开关的优先选择包括：（i）照片以蓝色，远红色和近红外（NIR）范围（400-800 nm）发射，以使现有的曲目多样化联络并显着提高其动态和动态特性（特定目标1）；（ii）FDA批准的药物（抗病毒药）和卧室（咖啡因及其代谢产物）有望减少转化的障碍应用程序（特定目标2）。这些可切换的APS将允许团队远程控制抗体 - 抗原在高临时和/或空间上以可逆的方式识别和操纵内源性目标解决。同时，团队将展示工程智能APS的急性和纤维素中生物过程的精确启动和终止，以及体内免疫或远程人类疾病的啮齿动物和果蝇模型中的神经调节。引人入胜的初步数据我们被要求证明拟议的新方法的高可行性以及团队的掌握应用程序中描述的方法，测定和模型的曲目。创新该项目要生成的分子工具包将提供广泛的精确开关以启用许多未来的生物学问题，对生物医学领域产生了很高的可持续影响。