Project 3: Myeloid-lymphoid cell crosstalk in HNSCC therapy

项目摘要 一些头颈部鳞状细胞癌（HNSCC）患者在治疗后观察到明显的临床反应， 免疫干预这些成功证明了治疗（重新）激活的力量 抗肿瘤T细胞来控制癌症，但考虑到更多的患者没有 当提供相同的治疗时体验临床益处。越来越多的证据表明， 免疫疗法对HNSCC的成功：（i）肿瘤的抗原性通常很差，这限制了免疫疗法的产生。 抗肿瘤免疫，（ii）抑制机制，加强肿瘤内的免疫耐受。P01的 目标是解决这两个障碍：项目1和2旨在确定增加HNSCC抗原性的方法; 该项目旨在将增强的抗原性引入成功的抗肿瘤免疫应答。为此我们 将利用骨髓细胞群及其与T细胞的相互作用。当考虑骨髓细胞时，我们 特别感兴趣的是肿瘤浸润树突状细胞（DC）群体，因为它们可以呈递肿瘤抗原 T细胞和许可证的执行T细胞介导的肿瘤控制。我们将首先评估三种药物， 我们根据它们在临床中的使用、它们与其他项目的相关性、我们的初始数据以及 它们靶向多种肿瘤相关成分的能力。即，我们将研究：（1）5 '氮杂胞苷，因为它 增强HNSCC患者和小鼠中的肿瘤细胞抗原性（另见项目1）;（2）抗PD-1 mAb，因为它 可以激活T细胞产生关键的抗肿瘤细胞因子，如IFN-γ，并在一些HNSCC中有效 （3）激动性抗CD 40 mAb，因为它可以诱导DC亚群产生效应细胞因子IL-12， 其促进抗肿瘤T细胞活性。因此，这些药物可以合理组合，以促进肿瘤 通过同时靶向肿瘤细胞、适应性免疫细胞和先天性免疫细胞进行控制。我们还将分析其他 在5年项目期间，部分根据项目1和2的调查结果，定义药物对 我们将利用正在进行的临床试验和动物模型， 人类疾病的主要特征。我们还将使用两种互补的单细胞分辨率方法： 单细胞RNA测序（scRNAseq）和单细胞成像（scIMAG）。scRNAseq将提供一个公正的 在人类和小鼠病变中观察药物诱导的免疫变化，并允许在物种间进行比较， 促进HNSCC转化研究。scIMAG将提供有关药物诱导免疫的信息， 随着时间的推移和肿瘤的地理背景下的反应。我们假设这些方法 可以确定药物诱导的髓系淋巴细胞串扰的机制，这是因果关系， HNSCC控制，并可用于治疗。我们应该做好准备， 工作，有研究免疫细胞和免疫治疗药物的经验，并验证了工具 将在这个项目中得到利用。此外，我们还组建了一个专家小组， 通过提供HNSCC免疫治疗，细胞分析，成像和统计学方面的关键专业知识，

PROJECT SUMMARY Some head and neck squamous cell carcinoma (HNSCC) patients see pronounced clinical responses with immunotherapeutic intervention. These successes demonstrate the power of therapeutically (re-)activating antitumor T cells to control cancer, but are insufficient, considering the many more patients who do not experience clinical benefit when provided the same treatments. Increasing evidence shows two main barriers to immunotherapy’s success against HNSCC: (i) the tumor’s often poor antigenicity, which limits the generation of antitumor immunity, (ii) suppressive mechanisms that enforce immune tolerance within tumors. This P01’s goal is to address these two barriers: Projects 1 and 2 aim to identify ways that increase HNSCC antigenicity; this Project aims to direct enhanced antigenicity into a successful antitumor immune response. To this end, we will harness myeloid cell populations and their crosstalk with T cells. When considering myeloid cells, we are particularly interested in tumor-infiltrating dendritic cell (DC) populations, since they can present tumor-antigen to T cells and license the execution of T cell-mediated tumor control. We will initially assess three drugs, which we carefully chose based on their use in the clinic, their relevance to the other Projects, our initial data, and their ability to target multiple tumor-associated components. Namely, we will study: (1) 5’azacytidine since it augments tumor cell antigenicity in HNSCC patients and mice (see also Project 1); (2) anti-PD-1 mAb since it can activate T cells to produce key antitumor cytokines such as IFN-gamma and is efficacious in some HNSCC patients; (3) agonistic anti-CD40 mAb since it can induce a DC subset to produce the effector cytokine IL-12, which fosters antitumor T cell activity. Therefore, these drugs may be rationally combined to promote tumor control by targeting tumor cells, adaptive and innate immune cells simultaneously. We will also analyze other drugs during the 5-year project, based in part on findings from Projects 1 and 2. To define drug effects on myeloid⟷lymphoid cell crosstalk, we will leverage an ongoing clinical trial and animal models that recapitulate key features of the human disease. We will also use two complementary single cell resolution approaches: single cell RNA sequencing (scRNAseq) and single cell imaging (scIMAG). scRNAseq will provide an unbiased view of drug-induced immune changes in human and mouse lesions, and permit comparison across species to facilitate HNSCC translational research. scIMAG will provide information about drug-induced immune responses over time and within the geographical context of the tumor. We hypothesize that these approaches can identify mechanisms of drug-induced myeloid⟷lymphoid cell crosstalk, which are causally linked to HNSCC control and can be harnessed for therapy. We should be well positioned to perform the proposed work, having experience with studying immune cells and immunotherapy drugs, and having validated the tools that will be exploited in this project. Also, we have assembled a team of experts who will contribute to the project by providing key expertise in HNSCC immunotherapy, cell profiling, imaging, and statistics.