Bioengineering programmable and drug-controllable synthetic receptors fortunable CAR-T cell behaviors

生物工程可编程和药物可控合成受体可调节 CAR-T 细胞行为

• 批准号: 10383140
• 负责人: Nicholas Antonios Kalogriopoulos
• 金额: $ 6.76万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-15 至 2024-06-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10383140
• 关键词: Address; Adhesions; Antigens; Attention; B lymphoid malignancy; Behavior; Binding; Biomedical Engineering; CAR T cell therapy; Cells; Clinic; Clinical Research; Coin; Coupled; Custom; Dangerousness; Directed Molecular Evolution; Disease remission; Drug Controls; Drug Regulations; Engineering; Equilibrium; Face; Fortune; G-Protein-Coupled Receptors; Goals; Hyperactivity; Malignant Neoplasms; Membrane; Patient Care; Patients; Peptide Hydrolases; Pharmaceutical Preparations; Protein Engineering; Receptor Activation; Regulation; Relapse; Resolution; Safety; Signal Transduction; System; T cell response; T-Cell Activation; T-Lymphocyte; Testing; Therapeutic antibodies; Tissues; Toxic effect; Translations; Work; antigen detection; base; cell behavior; chimeric antigen receptor; chimeric antigen receptor T cells; cytokine; cytokine release syndrome; exhaustion; experimental study; high throughput screening; improved; neoplastic cell; next generation; notch protein; novel therapeutic intervention; prevent; programs; receptor; receptor expression; response; side effect; small molecule; standard of care; success; therapy resistant; transcription factor; tumor

Project Summary Chimeric Antigen Receptor (CAR)-T cell therapy has had unprecedented success in patients with B- cell malignancies, demonstrating upwards of 100% complete remission rate in some cases. Nearly all these patients, however, subsequently relapse with therapy-resistant tumors caused by antigen escape. In addition, the widespread utility of CAR-T cell therapy has been met with additional safety and efficacy challenges, including on-target/off-tumor toxicity, cytokine release syndrome caused from hyperactive CAR- T cells, and T cell exhaustion. A precise, well-balanced, and controlled CAR-T cell response is required to navigate these obstacles, avoid non-cancerous bystander tissues, limit dangerous side effects, and still identify and destroy tumor cells. New therapeutic approaches that permit the user-controlled, fine-tuned regulation of cell-autonomous CAR-T cell activity are needed to tune the CAR-T cell response to maintain this balance. To address these needs, and through the experiments outlined in this proposal, I will create a system for programmable and drug-controllable antigen-dependent cellular response coined PAGER (Programmable Adhesion GPCR-based, Exogenously Regulated). To further meet these needs, I will also add drug controllability to existing CARs. To create PAGER, I will engineer an adhesion G Protein-Coupled Receptor (GPCR) so that its structural rearrangements upon activation are coupled to the activity of a fused protease, allowing it to release a transcription factor upon receptor activation. Furthermore, to add drug- controllability to both PAGER and existing CARs, I will add a druggable protease-dependent degradation signal to synthetic receptors to create “drug-on” tunable PAGERs and CARs. I will utilize high-throughput screening, protein engineering, and directed evolution to accomplish these goals. Application of drug-controlled PAGERs and CARs in CAR-T cells will permit fine-tuned regulation of CAR-T cell activity to limit or prevent unwanted CAR-T cell responses. I will demonstrate proof-of-concept for using these synthetic receptors to precisely control customized CAR-T cell behaviors to an unprecedented resolution. PAGER will be used to control CAR expression and activity, cytokine secretion profile, and local delivery of therapeutic antibodies in a user-controlled cell-autonomous manner. The wide spectrum of cellular responses that can be programmed using PAGER highlights its great potential to assist in overcoming many of the safety and efficacy challenges that currently face CAR-T cell therapy.

项目摘要 嵌合抗原受体（CAR）-T细胞疗法在B-患者中取得了前所未有的成功 在某些情况下，细胞恶性肿瘤显示出100％的完全缓解率。几乎所有这些 然而，患者随后将抗原逃生引起的耐药性肿瘤传递。在 此外，已经满足了CAR-T细胞疗法的宽度效用，并具有额外的安全性和效率 挑战，包括靶向/肿瘤的毒性，由多动症引起的细胞因子释放综合征 T细胞和T细胞耗尽。需要精确，平衡和受控的CAR-T细胞响应才能 驾驶这些障碍，避免非癌性旁观者组织，限制危险的副作用，仍然 识别并破坏肿瘤细胞。允许用户控制的，微调的新治疗方法 需要调节细胞自主CAR-T细胞活性以调整CAR-T细胞反应以维持 这个平衡。 为了满足这些需求，并通过本提案中概述的实验，我将创建一个系统 用于可编程和可控制的抗原依赖性细胞反应引起的Pager （可编程粘附基于GPCR，外源调节）。为了进一步满足这些需求，我也会 将药物可控性添加到现有汽车中。要创建Pager，我将设计一个粘合剂G蛋白耦合 受体（GPCR），使其激活时的结构重排与融合的活性耦合 蛋白酶，使其能够在受体激活后释放转录因子。此外，要添加药物 - Pager和现有汽车的可控性，我将添加可吸毒的蛋白酶依赖性降解 向合成受体发出信号，以创建“药物”可调的打印机和汽车。我将使用高通量 筛选，蛋白质工程和指示进化以实现这些目标。 在CAR-T细胞中应用药物控制的寻呼机和汽车将允许对 CAR-T细胞活性以限制或防止不必要的CAR-T细胞反应。我将展示概念证明 用于使用这些合成受体来精确控制定制的CAR-T细胞行为 空前的解决方案。 Pager将用于控制汽车表达和活动，细胞因子分泌 概况，以及以用户控制的细胞自主方式的理论抗体的局部传递。宽 可以使用Pager进行编程的细胞响应范围突出显示其协助的巨大潜力 在克服目前面临CAR-T细胞疗法的许多安全性和有效挑战中。