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

项目总结 转移是癌症相关死亡的主要原因。传播到和殖民于 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)阐述了转移到远处需要系统免疫的中心假说 转移性淋巴结(LN)中免疫细胞和恶性细胞早期相互作用诱导的耐受。 在项目1中，我们已经成功地揭示了淋巴结转移的发生机制以及 这些转移就形成了远处转移，通过小鼠黑色素瘤模型，并已 在HNSCC的小鼠模型中展示了类似的现象。对于这次费用延期，我们将继续我们的 评估额外免疫亚群在黑色素瘤和HNSCC耐受过程中的作用，以及 确定肿瘤克隆异质性在原发肿瘤和LN转移中的作用。在项目2中，我们 已经成功开发了一种新的多重免疫荧光原位成像平台Codex，以 描述组织结构并演示CODEX在新鲜冷冻组织和FFPE组织中的应用 HNSCC和黑色素瘤的原代和小鼠样本。对于成本延长，项目2的目标是 继续开展工作，实现食典中RNA与蛋白质抗体的联合检测，以研究趋化因子和 在项目1和项目3中确定了候选细胞因子。在项目3中，我们开发了一种一体化的单细胞分析 研究我们中心产生的成像和测序数据的管道。通过这一分析，我们已经确定 与淋巴转移相关的潜在可药物趋化因子配体-受体对及其验证 与项目1团队合作的职能意义。我们完成的工作包括一台新奇的机器 学习细胞类型识别方法“Ceresta”，以促进对空间生物学的新见解。对于成本 扩展，项目3旨在继续应用我们的计算工具和分析管道来调查 额外的免疫细胞类型和基质细胞，以确定促进淋巴转移的组织结构的介体 HNSCC组织标本的转移。我们将在独立队列中验证这一发现的一致性 人类HNSCC TMA(核心3)，它带有生存结果的注释，并与项目1合作 功能研究。此外，我们的中心建议继续努力，通过以下方式进行外联和传播 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.