Spatial-Genomic Integrative Multi-Species Analysis of Lymph Node Metastasis

项目概要 转移是大多数癌症相关死亡的原因。传播和殖民化 在大多数实体瘤中，淋巴结（LN）先于远处转移。我们的癌症系统中心 生物学 (CCSB) 提出了一个中心假设，即远处转移需要全身免疫 由转移性淋巴结（LN）中免疫细胞和恶性细胞之间的早期相互作用诱导的耐受性。 在项目1中，我们成功揭示了淋巴结转移的机制以及 通过黑色素瘤小鼠模型，这些转移对远处转移形成的影响已被证实 在 HNSCC 小鼠模型中展示了类似的现象。为了延长成本，我们将继续我们的 致力于评估其他免疫亚群在黑色素瘤和 HNSCC 耐受过程中的作用，以及 确定原发肿瘤和淋巴结转移之间肿瘤克隆异质性的作用。在项目2中，我们 成功开发了新型多重免疫荧光原位成像平台 CODEX， 表征组织结构并演示 CODEX 在新鲜冷冻和 FFPE 组织中的应用 HNSCC 和黑色素瘤的原发样品和鼠样品。对于成本扩展，项目 2 旨在 继续努力在 CODEX 中实现 RNA 与蛋白质抗体的共同检测，以研究趋化因子和 项目 1 和项目 3 中确定的候选细胞因子。在项目 3 中，我们开发了综合单细胞分析 研究我们中心生成的成像和测序数据的管道。通过此分析，我们确定了 与淋巴结转移相关的潜在可药物趋化因子配体-受体对，并验证了它们的 与项目 1 团队合作的功能意义。我们完成的工作包括一台新颖的机器 学习细胞类型识别方法“CELESTA”，以促进对空间生物学的新见解。为了成本 扩展，项目 3 旨在继续应用我们的计算工具和分析管道进行调查 额外的免疫细胞类型和基质细胞来识别促进淋巴结的组织结构的介质 HNSCC 组织样本上的转移。我们将在独立队列中验证研究结果的一致性 人类 HNSCC TMA（核心 3），其注释有生存结果并与项目 1 合作 功能研究。此外，我们中心建议继续努力通过以下方式进行外展和传播： 虚拟每月研讨会。

PROJECT SUMMARY Metastasis is responsible for the majority of cancer-associated deaths. Dissemination to and colonization of lymph nodes (LNs) precedes metastasis to distant sites in most solid tumors. Our Center in Cancer Systems Biology (CCSB) addresses the central hypothesis that metastasis to distant sites requires systemic immune tolerance induced by early interactions between immune and malignant cells in the metastatic lymph nodes (LN). In Project 1, we have successfully uncovered mechanisms by which metastasis to LNs occurs and the effects of these metastases on the formation of distant metastases, through a murine model of melanoma and have been demonstrating similar phenomena in a murine model of HNSCC. For this cost extension, we will continue our work to evaluate the roles of additional immune subsets in the tolerance process in melanoma and HNSCC, and determine the roles of tumor clonal heterogeneity between primary tumors and LN metastases. In Project 2, we have successfully developed a novel multiplexed immunofluorescence in situ imaging platform, CODEX, to characterize tissue architecture and demonstrated the application of CODEX to fresh frozen and FFPE tissue samples of primary and murine samples of HNSCC and melanoma. For the cost extension, Project 2 aims to continue its work to achieve co-detection of RNA with protein antibodies in CODEX to study the chemokine and cytokine candidates identified in Projects 1 and 3. In Project 3, we developed an integrative single-cell analysis pipelines to study imaging and sequencing data generated in our center. Through this analysis we have identified potentially druggable chemokine ligand-receptor pairs related to lymph node metastasis and validated their functional significance in collaboration with the Project 1 team. Our completed work includes a novel machine learning cell type identification method “CELESTA” to facilitate new insights on spatial biology. For the cost extension, Project 3 aims to continue to apply our computational tools and analysis pipelines to investigate additional immune cell types and stroma cells to identify mediators of tissue architectures facilitating lymph node metastasis on HNSCC tissue samples. We will validate the consistency of the finding on an independent cohort of human HNSCC TMA (Core 3), which is annotated with survival outcomes and collaborate with Project 1 for functional studies. In addition, our Center proposes to continue our efforts in outreach and dissemination through virtual monthly seminars.