SaefCAR: Regulatable CAR-T cells for safe and effective immunotherapy

SaefCAR：可调节的 CAR-T 细胞用于安全有效的免疫治疗

• 批准号: 10759600
• 负责人: Yemi Onakunle
• 金额: $ 40万
• 依托单位: MABSWITCH, INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-19 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10759600
• 关键词: Address; Adverse event; Affinity; Animal Model; Antibodies; Antigen Targeting; Antigens; Architecture; Binding; Biological Products; CAR T cell therapy; CD19 gene; Calmodulin; Cell Line; Cells; Clinical; DNA; Discrimination; Disease; Dose; Engineering; Equilibrium; FDA approved; Flow Cytometry; Future; Hematologic Neoplasms; Human; Immunoglobulin Fragments; Immunotherapy; In Vitro; Letters; Libraries; Ligands; Modality; Patients; Peptides; Phase; Physiological; Proteins; Reaction Time; Regulation; Reporter; Retroviral Vector; Risk; Safety; Serious Adverse Event; Signal Transduction; Signal Transduction Induction; Small Business Innovation Research Grant; Solid Neoplasm; Specificity; T-Cell Activation; T-Lymphocyte; Technology; Tissues; Toxic effect; Tumor Antigens; Tumor Lysis Syndrome; Variant; Work; Writing; antigen binding; cancer immunotherapy; cancer type; chimeric antigen receptor; chimeric antigen receptor T cells; clinical translation; cytokine; cytokine release syndrome; delivery vehicle; design; engineered T cells; exhaustion; experience; improved; in vivo; neurotoxicity; preclinical development; premature; prevent; screening; small molecule; success; systemic toxicity; tumor

Project Summary: Despite the remarkable success in the treatment of some hematological cancers, the risk of serious adverse events and loss of potency remain major problems that threaten to curtail widespread application of chimeric antigen receptor (CAR) T-cell therapies to other cancer types including solid tumors. These problems, all of which are influenced by the binding affinity of the CAR targeting domain, result from the propensity for unpredictable hyperactivation that can lead to potentially fatal systemic toxicities such as cytokine release syndrome (CRS), tumor lysis syndrome (TLS) and neurotoxicity. Furthermore, on-target off-tumor toxicities can result from the indiscriminate binding of constitutively expressed CARs to tumor antigens on healthy tissues, whilst disease relapses can occur due to antigen escape and/or T cell exhaustion. Current and emerging CAR- T cell strategies and designs to address these problems are often too slow in terms of response times and onset of action. Furthermore, they prematurely and permanently eliminate the “high value” cells and often require the expression of additional proteins which may constrain the DNA payload capacity of current lenti- and retro- virus vectors. There is thus an urgent unmet need for in-vivo regulatable CAR-T cell platforms that are rapid, reversible and don’t exert a substantial DNA burden on the CAR delivery vector. The objective of this proposal is to assess the feasibility to regulate the activation, effector functions and cytokine release of CAR T-cells using an exogenously administered ligand that modulates the affinity of the CAR T-cell targeting domain. To this end we will utilize our proprietary universal allosteric-linker and switch module for antibodies (UNASMA) technology to incorporate an ON/OFF affinity switch into a well characterized antibody single chain variable fragment (scFv) against a clinically validated hematological cancer target which will subsequently be formatted as a switchable affinity chimeric antigen receptor (SaefCAR). We postulate that the resulting SaefCAR following transduction into a T-cell, will enable the in-vivo regulation of the SaefCAR T-cells’ activity via exogenous administration of a small molecule or peptide ligand, potentially solving several of the problems of current CAR-T cell therapies by: 1) Ameliorating intrinsic toxicities via enablement of a slow start to T-cell activation, 2) Preventing on-target off- tumor toxicities by switching off the CAR-T cell without eliminating it 3) Preventing exhaustion of tumor-specific CAR T cells caused by tonic signaling and other mechanisms via intermittent ON/OFF switching of the CAR. Specifically in Aim 1 we will develop switchable forms of the scFv which will subsequently be formatted into a CAR and transduced into a T-cell in Aim 2. In Aim 3 we will validate proof-of-concept of the regulated SaefCAR T-cells in in-vitro and in-vivo studies. We aim to demonstrate that a ligand-controllable affinity switch in the binding domain of SaefCAR T-cells can regulate it’s activation and effector functions against a tumor. Importantly due to the universal character of our approach, it provides a blueprint to improve any scFv-based CAR T-cell therapy in the future by adding a capability to regulate the CAR affinity for the tumor antigen.

项目摘要： 尽管在治疗某些血液学癌症方面取得了显着成功，但严重逆境的风险 事件和效力丧失仍然是有可能减少嵌合宽度应用的主要问题 抗原受体（CAR）T细胞疗法对包括实体瘤在内的其他癌症类型。这些问题，所有 受汽车靶向域的结合亲和力的影响，这是由于对 不可预测的过度激活可能导致潜在的致命全身毒性，例如细胞因子释放 综合征（CRS），肿瘤裂解综合征（TLS）和神经毒性。此外，靶向非肿瘤毒性可以 由组成型表达的汽车与健康组织上的肿瘤抗原的不加选择的结合导致 而疾病继电器可能是由于抗原逃生和/或T细胞耗尽而发生的。电流和新兴汽车 - 在响应时间和发作方面，解决这些问题的T细胞策略和设计通常太慢了 行动。此外，它们几乎可以永久消除“高价值”细胞，并且经常需要 表达其他蛋白质的表达可能会限制当前扁豆和恢复病毒的DNA有效载荷能力 向量。因此，紧急无法满足的对体内可调节的CAR-T细胞平台的需求快速，可逆 并且不要在汽车输送载体上施加大量的DNA燃烧。该提议的目的是评估 使用A的激活，效应子功能和细胞因子释放CAR T细胞的可行性 外源给予的配体调节CAR T细胞靶向结构域的亲和力。为此，我们 将利用我们专有的通用变构链链和开关模块用于抗体（UNASMA）技术 将ON/OFF亲和力开关纳入表征良好的抗体单链可变片段（SCFV） 针对经过临床验证的血液学癌症靶标，后来将以可切换格式 亲和力嵌合抗原受体（SAEFCAR）。我们假设由此产生的SAEFCAR转移后 进入T细胞，将通过外源给药saefcar T细胞的活性进行体内调节 小分子或胡椒配体，可能通过以下方式解决当前CAR-T细胞疗法的几个问题 1）通过使T细胞激活缓慢地启用固有毒性，2）防止目标远离目标 通过关闭CAR-T细胞而无需消除肿瘤的毒性3）防止肿瘤特异性 由滋补信号传导和其他机制引起的CAR T细胞通过间歇性打开/关闭汽车的开关。 特别是在AIM 1中，我们将开发SCFV的可切换形式，然后将其格式化为 汽车并在AIM 2中翻译成T细胞。在AIM 3中，我们将验证受监管的SAEFCAR的概念证明 体外和体内研究中的T细胞。我们的目的是证明该配体可控制的亲和力开关 SAEFCAR T细胞的结合结构域可以调节其激活和效应子功能针对肿瘤。重要的是 由于我们方法的普遍特征，它提供了一个蓝图来改善任何基于SCFV的汽车T细胞 将来通过增加调节对肿瘤抗原的汽车亲和力的能力来进行治疗。