Project 3: Myeloid-lymphoid cell crosstalk in HNSCC therapy

项目总结 一些头颈部鳞状细胞癌(HNSCC)患者看到明显的临床反应 免疫治疗干预。这些成功证明了治疗(重新)激活的力量。 抗肿瘤T细胞可以控制癌症，但考虑到更多不能控制癌症的患者，这是不够的 当提供相同的治疗时，体验临床益处。越来越多的证据表明有两个主要障碍 免疫疗法对HNSCC的成功：(I)肿瘤的抗原性往往很差，这限制了生成 抗肿瘤免疫，(Ii)在肿瘤内加强免疫耐受的抑制机制。本P01‘S 目标是解决这两个障碍：项目1和项目2的目标是确定提高HNSCC抗原性的方法； 该项目旨在将增强的抗原性引导到成功的抗肿瘤免疫反应中。为此，我们 将利用髓系细胞群体及其与T细胞的串扰。当考虑髓系细胞时，我们是 对肿瘤浸润性树突状细胞(DC)群体特别感兴趣，因为它们可以提呈肿瘤抗原 T细胞，并许可执行T细胞介导的肿瘤控制。我们将对三种药物进行初步评估， 我们根据它们在临床中的使用情况、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。也就是说，我们将研究：(1)5‘氮杂胞苷 增强HNSCC患者和小鼠的肿瘤细胞抗原性(另见项目1)；(2)抗PD-1单抗 能激活T细胞产生关键的抗肿瘤细胞因子，如干扰素-γ，对某些HNSCC有效 患者；(3)激动型抗CD40单抗，因为它可以诱导DC亚群产生效应细胞因子IL-12， 培养抗肿瘤的T细胞活性。因此，这些药物可能会合理组合，以促进肿瘤 通过同时靶向肿瘤细胞、适应性细胞和先天免疫细胞进行控制。我们还将分析其他 五年项目期间的药物，部分基于项目1和项目2的结果。 髓系⟷淋巴细胞串扰，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元格解析方法： 单细胞RNA测序(ScRNAseq)和单细胞成像(ScIMAG)。ScRNAseq将提供不偏不倚的 查看药物诱导的人类和小鼠损伤的免疫变化，并允许跨物种比较 促进HNSCC的翻译研究。SCIMAG将提供有关药物诱导免疫的信息 随着时间的推移和在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⟷淋巴样细胞串扰的机制，这些机制与 HNSCC控制，可用于治疗。我们应该能够很好地执行提议的 工作，有研究免疫细胞和免疫疗法药物的经验，并验证了这些工具 这一点将在这个项目中得到开发。此外，我们还组建了一个专家团队，他们将为 该项目提供HNSCC免疫治疗、细胞图谱、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.