New regulators of natural killer cell-mediated cytotoxicity

项目摘要 自然杀伤（NK）细胞是先天免疫系统中的关键效应淋巴细胞。它们展现所 检测和清除转化细胞和病毒感染细胞的显著能力。NK细胞也是一个关键 在同种异体供体移植器官排斥反应中的作用。NK细胞介导的杀伤的不平衡是 与主要形式的人类疾病如癌症、慢性感染和免疫缺陷相关。到 为了提高基于NK细胞的疗法的疗效，关键是要确定控制NK细胞应答的基因。 靶细胞对NK细胞介导的杀伤。在我们的初步研究中， 来剖析NK样细胞系对靶细胞的杀伤。这些筛选分离出了已知的NK细胞杀伤调节因子， 大多数的命中先前与NK细胞功能无关。该试点项目的主要目标是 使用人原代NK细胞验证和表征这些候选基因。这些基因中的每一个都会 使用CRISPR基因组编辑在靶细胞中单独删除。然后我们将确定淘汰赛 NK细胞对原代NK细胞的杀伤有反应。一旦候选基因得到验证，我们将进一步表征 其在NK细胞介导的细胞毒作用中的作用和机制。该试点项目的成功完成将 大大拓宽了我们对NK细胞如何识别和杀死靶细胞的知识。从以下方面收集的见解 这项工作将有助于提高癌症和病原体感染免疫疗法的有效性和安全性。

PROJECT SUMMARY Natural killer (NK) cells are key effector lymphocytes in the innate immune system. They exhibit the remarkable abilities of detecting and eradicating transformed and virus-infected cells. NK cells also play a key role in the rejection of transplanted organs from allogeneic donors. Imbalances in NK cell-mediated killing are associated with major forms of human disease such as cancer, chronic infection, and immunodeficiency. To improve the efficacy of NK cell-based therapies, it is critical to identify the genes controlling the response of target cells to NK cell-mediated killing. In our preliminary studies, we performed genome-wide genetic screens to dissect target cell killing by an NK-like cell line. The screens isolated known regulators of NK cell killing but the majority of the hits were not previously linked to NK cell functions. The major goal of this pilot project is to validate and characterize these candidate genes using human primary NK cells. Each of these genes will be individually deleted in target cells using CRISPR genome editing. We will then determine how the knockout cells respond to killing by primary NK cells. Once a candidate gene is validated, we will further characterize its function and mechanism in NK cell-mediated cytotoxicity. Successful completion of this pilot project will substantially broaden our knowledge of how NK cells recognize and kill target cells. Insights gleaned from this work will help improve the efficacy and safety of immunotherapies for cancer and pathogen infection.