Single Cell Genome-Wide Myeloid Response Profiling in Immunotherapy

项目总结 肿瘤微环境是多种免疫细胞类型的家园，但目前的免疫治疗方法 主要集中在激活细胞溶解T细胞上。其他髓系细胞在抗癌中的潜力 到目前为止，对它们的探索要少得多。例如，我们对髓系细胞的复杂性了解有限。 细胞亚型，我们不能完全区分肿瘤促进细胞和肿瘤抑制细胞，而且我们缺乏 关于可用于治疗的已定义的髓系细胞相关分子通路的信息。 在这里，我们将使用尖端的、无偏见的单细胞图谱来揭示未被认可的免疫调节 髓系细胞类型，以及体外和体内的扰动，揭示未被发现的肿瘤浸润性 髓系(TIM)细胞群并确定它们在肺癌中的功能作用。为此，我们将首先确定 人类TIM状态，它们与临床参数的相关性，以及这些状态是否在 人和小鼠肺腺癌。我们将具体测试以下假设：i)尚未被赏识 人类肿瘤和患者外周血中的髓系细胞状态与患者的生存相关(和 可能是其他临床参数)，以及ii)肺的基因工程小鼠肿瘤模型 腺癌宿主保存了人类TIM状态，并证明了进一步的动物研究这些状态的功能 各州。其次，考虑到我们的初始数据，我们将重点关注所谓的GN2和GN3中性粒细胞亚群 显示它们在人和小鼠肺肿瘤中的存在及其与癌症进展的相关性 (《科学》，2017年出版，我们的未发表数据包含在本申请中)，中性粒细胞 成为各种实体肿瘤，尤其是肺癌患者存活的有力预测指标。我们会 具体测试以下假设：1)GN2和GN3中性粒细胞具有不同的促癌功能，以及 Ii)这些亚群使用特定的分子信号通路来促进肺癌的进展。为了实现我们的目标 目标，我们将结合两个具有互补专业知识的实验室的努力：肿瘤免疫生物学和髓系 细胞(Pittet)、单细胞RNA测序(scRNA-Seq)和理论/生物信息学(Klein)。我们还有更多 与临床医生合作，从肺癌患者身上获取血液样本和新鲜肿瘤活检组织(我们的 初步数据也包括在本申请中)。总体而言，这里开发的方法和资源 可能会对开发新的、更有效的免疫疗法产生重大影响。此外，通过将目标对准 T细胞以外的免疫系统，我们将利用非冗余免疫组件的多样性作为一种方式 克服当前治疗方案的局限性。

Project summary Tumor microenvironments are home to diverse immune cell types but current immunotherapeutic approaches are focused largely on activating cytolytic T cells. The potential of other, myeloid lineage cells in fighting cancer are hitherto much less explored. For example, we have a limited understanding of the complexity of myeloid cell subtypes, we cannot fully discriminate between tumor-promoting and tumor-suppressing cells, and we lack information about defined myeloid cell-associated molecular pathways that could be harnessed for therapy. Here, we will use cutting-edge, unbiased single cell profiling to reveal unappreciated immunoregulatory myeloid cell types, alongside in vitro and in vivo perturbations, to reveal unappreciated tumor-infiltrating myeloid (TIM) cell populations and define their functional role in lung cancer. To this end, we will first determine human TIM states, their correlation with clinical parameters, and whether these states are conserved between human and mouse lung adenocarcinoma. We will specifically test the hypotheses that i) yet-unappreciated myeloid cell states in tumors and peripheral blood of human patients correlate with patient survival (and possibly other clinical parameters), and ii) genetically engineered mouse tumor models of lung adenocarcinoma host conserved human TIM states and justify further animal studies of the function of these states. Second, we will focus on so-called GN2 and GN3 neutrophil subsets, considering our initial data showing their existence in both human and mouse lung tumors and their relevance to cancer progression (Science, 2017 in Press and our unpublished data included in this application) and that neutrophils are emerging as strong predictors of survival for diverse solid tumors and most notably lung cancer. We will specifically test the hypotheses that i) GN2 and GN3 neutrophils have distinct tumor-promoting functions, and ii) these subsets use specific molecular signaling pathways to foster lung cancer progression. To achieve our goals, we will combine efforts of two labs with complementary expertise: tumor immunobiology and myeloid cells (Pittet), and single cell RNA sequencing (scRNA-Seq) and theory/bioinformatics (Klein). We have further teamed up with clinicians to obtain both blood samples and fresh tumor biopsies from lung cancer patients (our preliminary data are also included in this application). Overall, the approaches and resources developed here could have major implications for developing new and more efficient immunotherapies. Also, by targeting the immune system beyond T cells, we will exploit the diversity of non-redundant immune components as a way to overcome limitations of current treatment options.