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

项目摘要 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）先于远处部位转移。我们的癌症系统中心 生物学（CCSB）解决了中心假设，即转移到远处的网站需要全身免疫 在转移性淋巴结（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。 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， 通过黑色素瘤的鼠模型，这些转移对远处转移的形成有影响， 在HNSCC的鼠模型中证明了类似的现象。对于这一成本延长，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， 确定原发肿瘤和LN转移瘤之间肿瘤克隆异质性的作用。在项目2中，我们 成功开发了一种新型的多路免疫荧光原位成像平台CODEX， 表征组织结构，并证明CODEX在新鲜冷冻和FFPE组织中的应用 HNSCC和黑素瘤的原代和鼠样品。对于费用扩展，项目2旨在 继续其工作，以实现共同检测RNA与蛋白抗体在CODEX研究趋化因子， 在项目1和3中鉴定的细胞因子候选物。在项目3中，我们开发了一种整合的单细胞分析方法， 研究我们中心生成的成像和测序数据的管道。通过分析，我们发现 与淋巴结转移相关的潜在药物化趋化因子配体-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个新颖的机器 学习细胞类型识别方法“CELESTA”，以促进对空间生物学的新见解。费用 作为扩展，项目3旨在继续应用我们的计算工具和分析管道来研究 其他免疫细胞类型和基质细胞以鉴定促进淋巴结转移的组织结构的介质 HNSCC组织样品中的转移。我们将在一个独立的队列中验证该发现的一致性 人类HNSCC TMA（核心3），其中注释了生存结局，并与项目1合作， 功能研究。此外，本中心建议继续努力通过以下方式开展外联和传播工作： 每月虚拟研讨会。

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.