权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

De Novo Engineering of Small Molecule-Actuatable Biosensors for Cell Therapy

用于细胞治疗的小分子可驱动生物传感器的从头工程

基本信息

批准号：
10461083
负责人：
Liangcai Gu
金额：
$ 38.38万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-01 至 2024-07-31
项目状态：
已结题

项目摘要

De Novo Engineering of Small Molecule-Actuatable Biosensors for Cell Therapy ABSTRACT Preceded by small-molecule drugs and biologics, cell-based therapies, such as the use of chimeric antigen receptor T (CAR-T) cells against blood cancers, is becoming a new pillar of medicine. However, compared with traditional drugs, they are more susceptible to safety concerns due to difficulties associated with controlling cell actions in a therapeutic setting. My research program aims to develop a novel approach to engineer genetically encoded biosensors for spatiotemporal control of cell behaviors. We focus on the de novo engineering of the biosensors for using small molecules to control cell therapeutic responses, proliferation, death, migration, communication, and metabolism. Our strategy is to design chemically induced dimerization (CID) systems, in which two proteins dimerize only in the presence of a small molecule. To date, only a few CID systems are available and their dimerization inducers are not ideal for clinical use. The de novo design of CID systems with desired affinity and specificity for given small molecules remains an unsolved problem in the field of protein engineering. We will solve this problem by coupling computational protein design to our recently developed single-molecular-interaction sequencing (SMI-seq) technology. SMI-seq has the potential to break a key barrier to CID engineering by enabling large-scale, two-dimensional (or ‘library-by-library’) screening of two CID binder variant libraries. We will apply two parallel approaches to engineer CID systems: i) targeted screening of computationally designed binder libraries, and ii) random screening of vastly diverse combinatorial binder libraries (>109) using immunoglobulin or de novo designed scaffolds. We will assess the success rates, turnaround times, and cost-effectiveness of both approaches by testing a set of clinically approved antiviral drugs with excellent intracellular delivery efficiency as CID inducers. Finally, we will demonstrate the use of designed biosensors to control cellular processes in both cultured cells and a mouse model. Successful completion of this research will open up new possibilities for engineering ligand-responsive protein assemblies, an unexplored territory of protein design. Designed CID systems will significantly expand the chemogenetic toolkit for gene- and cell-based therapies, as well as systems and synthetic biology research.

用于细胞治疗的小分子可驱动生物传感器的从头工程抽象的先于小分子药物和生物制剂、基于细胞的疗法，例如使用嵌合抗原 T受体（CAR-T）细胞对抗血癌，正在成为医学的新支柱。然而，与传统药物，由于控制细胞的困难，它们更容易受到安全问题的影响治疗环境中的行动。我的研究计划旨在开发一种新的基因工程方法用于细胞行为时空控制的编码生物传感器。我们专注于从头工程使用小分子控制细胞治疗反应、增殖、死亡、迁移的生物传感器，通讯、新陈代谢。我们的策略是设计化学诱导二聚（CID）系统，其中两种蛋白质仅在小分子存在的情况下二聚化。迄今为止，只有少数 CID 系统可用，并且它们的二聚化诱导剂对于临床使用并不理想。 CID 系统的从头设计对给定小分子的所需亲和力和特异性仍然是蛋白质领域中尚未解决的问题工程。我们将通过将计算蛋白质设计与我们最近开发的单分子相互作用测序（SMI-seq）技术。 SMI-seq 有可能破解密钥通过实现大规模、二维（或“逐个库”）筛选两个库，消除了 CID 工程的障碍 CID 结合物变体库。我们将应用两种并行方法来设计 CID 系统： i) 有针对性筛选计算设计的结合物库，以及 ii) 随机筛选多种多样的组合使用免疫球蛋白或从头设计的支架的结合物文库（>109）。我们将评估成功率，通过测试一组临床批准的抗病毒药物来了解两种方法的周转时间和成本效益作为 CID 诱导剂具有优异的细胞内递送效率的药物。最后，我们将演示使用设计了生物传感器来控制培养细胞和小鼠模型中的细胞过程。成功的这项研究的完成将为工程配体响应蛋白质组装开辟新的可能性，蛋白质设计的一个未探索的领域。设计的 CID 系统将显着扩展化学遗传学用于基于基因和细胞的疗法以及系统和合成生物学研究的工具包。