Project 1 Mouse Models Analysis

摘要/摘要：项目1 在大多数实体恶性肿瘤中，淋巴结(LN)转移先于进一步扩散，并可定为III期。 诊断。尽管如此，LN转移在进一步疾病进展中的机制作用很差。 明白了。我们在小鼠身上发现，在定植LNS时，肿瘤细胞激活转录程序， 通过与LN白细胞的相互作用，使其能够诱导肿瘤特异性免疫耐受。这些 随后，白细胞在宿主体内循环，导致全身免疫耐受。 远处转移性播种。重要的是，我们在LN中发现了相同的保守转录签名 人类的转移，提示我们的LN转移和免疫耐受的机制 小鼠模型与人类癌症有关。我们假设肿瘤细胞之间的局部相互作用， 在LN定植之前、期间和之后，LN内的白细胞和间质改变远处组织以促进 疾病的发展。我们预计这些相互作用将涉及同型和异型细胞-细胞相互作用， 协调LN、肿瘤和远程站点内的架构变化，以及各种人口的贩运 在整个宿主中。我们将使用同基因小鼠模型来确定这些相互作用的机制 通过以下方法促进疾病的发展。1：确定肿瘤的动力学 产生系统免疫耐受性。使用高维成像平台(CODEX)，我们将揭开 整个宿主的局部微环境的纵向变化。2：询问异型细胞的本质--细胞 LNS内的相互作用及其促进免疫逃避和诱导的机制 免疫耐受性。我们将使用scrna-seq来识别免疫和间质的转录变化。 在淋巴结内与恶性细胞相互作用的群体。利用空间转录学，我们将确定 与恶性细胞相互作用的细胞的基因表达模式与远距离的不同。要确认 这些确定的目标和项目2中确定的目标的功能意义，我们将结合CRISPR- 转移性LN肿瘤的介导性基因编辑以及细胞耗竭研究和基因敲除小鼠或诱导型 老鼠模型。3：确定来自肿瘤相关淋巴结的免疫细胞的运输如何建立促进转移的 远距离场地的微环境。我们将使用一个新的细胞标记平台来评估贩运 白细胞和间质细胞从肿瘤累及的淋巴结转移到远处，再回到原发肿瘤。这些 研究将揭示LN转移促进肿瘤向远处扩散的机制，并确定 防治转移性疾病的新靶点。

SUMMARY/ABSTRACT: PROJECT 1 Lymph node (LN) metastasis precedes further dissemination for most solid malignancies and confers a stage III diagnosis. Nonetheless, the mechanistic role of LN metastasis in further disease progression is poorly understood. We discovered in mice that in colonizing LNs, tumor cells activate transcriptional programs that enable their induction of tumor-specific immune tolerance through interactions with LN leukocytes. These leukocytes subsequently recirculate throughout the host, resulting in systemic immune tolerance facilitating metastatic seeding of distant sites. Importantly, we found the same conserved transcriptional signature in LN metastases in humans, suggesting that the mechanisms driving LN metastasis and immune tolerance in our mouse model are pertinent to human cancers. We hypothesize that local interactions between tumor cells, leukocytes, and stroma within LNs before, during, and following LN colonization alter distant tissues to promote disease progression. We expect these interactions to involve homotypic and heterotypic cell-cell interactions, orchestration of architectural changes within LNs, tumors, and distant sites, and trafficking of various populations throughout the host. We will use syngeneic mouse models to identify the mechanisms by which these interactions facilitate disease progression through the following approaches. 1: Determine the kinetics by which tumors generate systemic immune tolerance. Using a high-dimensional imaging platform (CODEX), we will uncover longitudinal changes in local microenvironments across the host. 2: Interrogate the nature of heterotypic cell-cell interactions within LNs and the mechanisms by which these interactions facilitate immune evasion and induction of immune tolerance. We will use scRNA-seq to identify transcriptional changes in immune and stromal populations interacting with malignant cells within LNs. Using spatial transcriptomics, we will determine how the gene expression patterns of cells interacting with malignant cells differ from those at a distance. To confirm the functional significance of these targets identified and those identified in Project 2, we will combine CRISPR- mediated gene editing of LN metastatic tumors, along with cell depletion studies and knockout mice or inducible mouse models. 3: Determine how trafficking of immune cells from tumor-involved LNs establish a pro-metastatic microenvironment at distant sites. We will employ a novel cell labeling platform to evaluate trafficking of leukocytes and stromal cells from tumor-involved LNs to distant sites and back to the primary tumor. These studies will reveal the mechanisms whereby LN metastases promote tumor spread to distant sites and identify novel targets for the prevention and treatment of metastatic disease.