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

项目摘要/摘要 自然杀伤(NK)细胞是一种先天免疫效应细胞，在宿主防御中起直接作用。这个 NK细胞的激活是由受体-配体相互作用和下游细胞内信号通路介导的。 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的。 受体(CARS)。它们是由靶标识别和细胞激活功能组成的工程化受体。 可以引导免疫细胞介导对癌细胞的杀伤的蛋白。虽然T细胞是主要的免疫细胞 用于汽车免疫治疗的细胞类型，NK细胞比汽车工程T细胞有显著优势 细胞，因为它们可以来自非自体来源。然而，大多数研究测试CAR-NK细胞 使用了基于T细胞信号通路的CAR结构，而这些信号通路并没有针对NK细胞信号进行优化。在……里面 此外，汽车结构的开发在很大程度上是通过试错实验方法实现的， 目前还没有系统地了解改变CAR信号域是如何影响细胞激活的。 这项建议的主要目标是利用系统生物学识别有效的基于自然语言的汽车设计 工具。我们的方法结合了计算建模和定量磷酸蛋白质组学来产生去- 对CAR介导的NK细胞信号转导和细胞毒作用的了解。我们工作的结果将是一套 靶向并杀死BCMA阳性多发性骨髓瘤(MM)癌细胞的有效NK CARS。我们还将测试 优化后的CARS对表达CS1的多发性骨髓瘤细胞有抑制作用。该方法建立在我们团队的丰富经验基础上 对细胞信号进行建模和表征，并研究NK细胞生物学。在强劲的初步数据的指引下， 我们建议追求三个具体目标：(1)表征CAR-1细胞内和细胞水平的反应。 表达NK细胞；(2)建立预测CAR表达动态反应的计算模型 (3)鉴定能有效激活NK细胞的新型CAR结构。 总而言之，我们提议的研究将对汽车信号如何编码产生一个量化的理解 NK细胞介导的细胞毒性以及如何优化NK-CAR结构用于癌症免疫治疗。我们的 研究将广泛影响癌症免疫治疗领域，提供对细胞内NK细胞如何 信号和CAR结构影响NK细胞的激活。最终，这项研究将扩大我们对 NK细胞信号转导与NK-CAR免疫治疗其他肿瘤类型和癌症的设计标准 抗原。我们的工作有可能解锁NK-CAR细胞对癌症免疫的转化能力- 心理治疗。

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.