A chemistry-driven approach to Senolytic Bispecific T-Cell Engagers

化学驱动的 Senolytic 双特异性 T 细胞接合剂方法

• 批准号: EP/Y024699/1
• 负责人: Gonçalo Bernardes
• 金额: $ 25.55万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Fellowship
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=EP%2FY024699%2F1
• 关键词: chemistry; driven; approach; Senolytic; Bispecific

Cellular senescence involves the stress-induced, durable cell-cycle arrest of previously replication-competent cells and has been associated with a plethora of age-related pathologies, such as liver and lung fibrosis, atherosclerosis, diabetes, and osteoarthritis.Since most small molecules with senolytic activity lack potency and produce substantial side effects, there is growing interest in the development of senolytic agents that selectively eliminate senescent over healthy cells. The present project will tackle this problem by establishing a new, chemistry-driven approach for the assembly of unique Bispecific T-Cell Engagers (BiTEs) with senolytic activity.This will be achieved by designing two orthogonal reactions that are specific to cysteine (one of them exclusively targeting N-terminal Cys and therefore allowing asymmetric ligation), for the assembly of a library of bispecific antibodies (bsAbs) from individual Cys-tagged monoclonal antibodies (mAbs) and avoiding the need to clone, express, purify and test each bsAbs one by one. The approach will then consist on expressing two sets of Cys-tagged antibodies, one targeting cell-surface antigens on hematopoietic and immune cells, and the other targeting senescent cells, which will be ligated to each other through designed linkers into bsAbs specific to senescent and T-cells antigens, and therefore capable to trigger an immune cytotoxic response towards senescent cells. To profile this library in vitro, the immunomodulatory properties of the senolytic BiTEs will be inferred from cell lysis and viability assays from cocultures of senescent cells and peripheral blood mononuclear cells. Following this, the immunomodulatory effects of selected bsAbs with senolytic activity will be elucidated using -omics and immunological tools. The technology developed will not be limited to bsAbs but will allow being extended to the ligation of any type of proteins, enabling future applications in protein-based drug discovery.

细胞衰老涉及应激诱导的、先前具有复制能力的细胞的持久细胞周期停滞，并与许多与年龄相关的病理学相关，例如肝和肺纤维化、动脉粥样硬化、糖尿病和骨关节炎。由于衰老活性缺乏效力并产生显着的副作用，人们对开发选择性消除衰老细胞而不是健康细胞的衰老活性剂越来越感兴趣。本项目将通过建立一种新的化学驱动方法来组装具有衰老活性的独特双特异性 T 细胞接合剂 (BiTE) 来解决这个问题。这将通过设计两个半胱氨酸特异性的正交反应来实现（其中之一）它们专门针对 N 端 Cys，因此允许不对称连接），用于从单个 Cys 标记的单克隆抗体 (mAb) 组装双特异性抗体 (bsAb) 文库，并避免需要对每一种双特异性抗体进行一一克隆、表达、纯化和测试。然后，该方法将包括表达两组 Cys 标记的抗体，一组针对造血细胞和免疫细胞上的细胞表面抗原，另一组针对衰老细胞，这两种抗体将通过设计的接头相互连接成针对衰老细胞和免疫细胞的双特异性抗体。 T 细胞抗原，因此能够触发针对衰老细胞的免疫细胞毒性反应。为了在体外分析该文库，将从衰老细胞和外周血单核细胞共培养物的细胞裂解和活力测定中推断出 senolytic BiTE 的免疫调节特性。此后，将使用组学和免疫学工具阐明具有衰老活性的选定 bsAb 的免疫调节作用。所开发的技术不仅限于双特异性抗体，还可以扩展到任何类型蛋白质的连接，从而实现未来在基于蛋白质的药物发现中的应用。