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，例如使用嵌合抗原受体T（CAR-T）细胞对抗血癌，正在成为医学的新支柱。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我的研究项目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细胞行为的从头工程。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，我们的策略是设计化学诱导二聚化（CID）系统，其中两个蛋白质仅在小分子存在下才二聚化，迄今为止，只有少数 CID 系统能够实现二聚化。现有的 CID 系统的二聚化诱导剂并不适合临床使用。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我们将通过将计算蛋白质设计与我们最近开发的技术相结合来解决这个问题。单分子相互作用测序（SMI-seq）技术有潜力突破一个关键。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 结合剂变体库。我们将应用两种并行方法来设计 CID 系统：i) 有针对性的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。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，设计的 CID 系统将显着扩展化学遗传学领域。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。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library.

通过组合单域抗体库的逐步噬菌体选择创建高度特异性的化学诱导蛋白质二聚化系统。

DOI：
发表时间：
2020
期刊：
影响因子：
0
作者：
Gomez;Watanabe, Kurumi;Jiang, Huayi;Kang, Shoukai;Gu, Liangcai
通讯作者：
Gu, Liangcai

Synthetic nanobody-functionalized nanoparticles for accelerated development of rapid, accessible detection of viral antigens.

合成纳米抗体功能化纳米粒子，用于加速开发快速、可访问的病毒抗原检测。

DOI：
发表时间：
2022-04-15
期刊：
影响因子：
12.6
作者：
Chen, Xiahui;Kang, Shoukai;Ikbal, Md Ashif;Zhao, Zhi;Pan, Yuxin;Zuo, Jiawei;Gu, Liangcai;Wang, Chao
通讯作者：
Wang, Chao

COMBINES-CID: An Efficient Method for De Novo Engineering of Highly Specific Chemically Induced Protein Dimerization Systems.

COMBINES-CID：高度特异性化学诱导蛋白质二聚化系统从头工程的有效方法。

DOI：
发表时间：
2019
期刊：
影响因子：
15
作者：
Kang, Shoukai;Davidsen, Kristian;Gomez;Jiang, Huayi;Fu, Xiaonan;Li, Zengpeng;Liang, Yu;Jahn, Molly;Moussa, Mahmoud;DiMaio, Frank;Gu, Liangcai
通讯作者：
Gu, Liangcai

Creating Red Light-Switchable Protein Dimerization Systems as Genetically Encoded Actuators with High Specificity.

创建红光可切换的蛋白质二聚化系统作为具有高特异性的基因编码致动器。