Modeling based design of chimeric antigen receptors for Natural Killer cell-based immunotherapy

项目总结/摘要 自然杀伤（NK）细胞是在宿主防御中发挥直接作用的先天免疫效应细胞。的 NK细胞的活化由受体-配体相互作用和下游细胞内信号传导途径介导， 的方式近年来取得巨大成功的一种免疫疗法是基于嵌合抗原 受体（汽车）。这些是由靶识别和细胞激活功能组成的工程受体， 它可以引导免疫细胞介导杀死癌细胞。虽然T细胞是主要的免疫细胞， 作为用于基于CAR的免疫治疗的细胞类型，NK细胞提供了优于CAR工程化T细胞的显著优势。 细胞，因为它们可以来自非自体来源。然而，大多数测试CAR-NK细胞的研究 已经使用了基于未针对NK细胞信号传导优化的T细胞信号传导途径的CAR构建体。在 此外，CAR构建体的开发主要是使用试错实验方法实现的， 并且对于改变CAR信号结构域如何影响细胞活化没有系统的理解。 该提案的主要目标是使用系统生物学来识别有效的基于NK的CAR设计 工具.我们的方法结合了计算建模和定量磷酸化蛋白质组学，以产生一个去磷酸化的蛋白质组学。 CAR介导的NK细胞信号传导和细胞毒性的尾式理解。我们的工作成果将是一套 经验证的NK汽车靶向并杀死BCMA阳性多发性骨髓瘤（MM）癌细胞。我们还将测试 针对表达CS 1的MM细胞的优化的汽车。该方法建立在我们团队丰富的经验基础上 细胞信号的建模和表征以及NK细胞生物学的研究。在强有力的初步数据的指导下， 我们提出了三个具体的目标：（1）表征CAR的细胞内和细胞水平的反应， （2）开发计算模型以预测CAR表达的动态响应; （3）鉴定有效激活NK细胞的新型CAR构建体。 总的来说，我们提出的研究将产生对CAR信号如何编码的定量理解。 NK细胞介导的细胞毒性以及NK-CAR构建体如何优化用于癌症免疫治疗。我们 这项研究将通过深入了解细胞内NK细胞是如何 信号传导和CAR结构影响NK细胞活化。最终，这项研究将扩大我们对 NK细胞信号传导和其他肿瘤类型和癌症的基于NK-CAR的免疫疗法的设计标准 抗原我们的工作有可能释放NK-CAR细胞对癌症免疫的变革力量。 疗法

PROJECT SUMMARY/ABSTRACT Natural killer (NK) cells are innate immune effector cells that play an immediate role in host defense. The activation of NK cells is mediated by receptor-ligand interactions and downstream intracellular signaling path- ways. One type of immunotherapy that has achieved great success in recent years is based on chimeric antigen receptors (CARs). These are engineered receptors composed of both target recognition and cell activation func- tions that can direct immune cells to mediate killing of cancer cells. Although T cells are the predominant immune cell type used for CAR-based immunotherapy, NK cells provide significant advantages over CAR-engineered T cells because they can be derived from non-autologous sources. However, most studies testing CAR-NK cells have used CAR constructs based on T cell signaling pathways that are not optimized for NK cell signaling. In addition, the development of CAR constructs is largely achieved using a trial-and-error experimental approach, and there is no systematic understanding of how altering the CAR signaling domains influences cell activation. The main objective of this proposal is to identify effective NK based-CAR designs using systems biology tools. Our approach combines computational modeling and quantitative phospho-proteomics to generate a de- tailed understanding of CAR-mediated NK cell signaling and cytotoxicity. The outcome of our work will be a set of validated NK CARs that target and kill BCMA-positive multiple myeloma (MM) cancer cells. We will also test the optimized CARs against CS1-expressing MM cells. The approach builds on our team’s extensive experience in modeling and characterizing cell signaling and studying NK cell biology. Guided by strong preliminary data, we propose to pursue three Specific Aims: (1) Characterize intracellular and cellular-level responses of CAR- expressing NK cells; (2) Develop computational models to predict the dynamic responses of CAR-expressing NK cells; (3) Identify novel CAR constructs that effectively activate NK cells. Collectively, our proposed research will generate a quantitative understanding of how CAR signaling encodes NK cell-mediated cytotoxicity and how NK-CAR constructs can be optimized for cancer immunotherapy. Our research will broadly impact the field of cancer immunotherapy by providing insight into how intracellular NK cell signaling and CAR structure influence NK cell activation. Ultimately, this research will expand our knowledge of NK cell signaling and the design criteria for NK-CAR-based immunotherapy for other tumor types and cancer antigens. Our work has the potential to unlock the transformative power of NK-CAR cells for cancer immuno- therapy.