Single Cell Genome-Wide Myeloid Response Profiling in Immunotherapy

项目摘要 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 主要集中在激活溶细胞性T细胞上。其他髓系细胞对抗癌症的潜力 迄今为止，很少有人探索。例如，我们对骨髓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和肿瘤抑制细胞，我们缺乏 关于可用于治疗的确定的骨髓细胞相关分子途径的信息。 在这里，我们将使用最先进的，无偏见的单细胞分析，以揭示未受到重视的免疫调节， 骨髓细胞类型，以及体外和体内扰动，以揭示未被认识到的肿瘤浸润 骨髓（TIM）细胞群，并确定其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。我们将具体测试假设，i）尚未赞赏 人类患者的肿瘤和外周血中的骨髓细胞状态与患者存活相关（并且 可能的其他临床参数），和ii）肺肿瘤的基因工程小鼠肿瘤模型 腺癌宿主保守的人TIM状态，并证明这些功能的进一步动物研究是合理的。 states.其次，考虑到我们的初始数据，我们将关注所谓的GN 2和GN 3中性粒细胞亚群 显示了它们在人类和小鼠肺肿瘤中的存在以及它们与癌症进展的相关性 （科学，2017年出版，我们未发表的数据包括在本申请中），中性粒细胞是 成为多种实体瘤（尤其是肺癌）生存的强有力预测因素。我们将 i）GN 2和GN 3嗜中性粒细胞具有不同的肿瘤促进功能，和 ii）这些亚群使用特定的分子信号传导途径来促进肺癌进展。实现我们 我们将联合收割机结合两个实验室的工作，它们具有互补的专业知识：肿瘤免疫生物学和骨髓 细胞（Pittet）和单细胞RNA测序（scRNA-Seq）和理论/生物信息学（Klein）。我们进一步 与临床医生合作，从肺癌患者身上获得血液样本和新鲜的肿瘤活检（我们的 初步数据也包括在本申请中）。总的来说，这里开发的方法和资源 可能对开发新的更有效的免疫疗法产生重大影响。此外，通过瞄准 除了T细胞之外的免疫系统，我们将利用非冗余免疫成分的多样性作为一种方法， 克服目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.