Project 1 Mouse Models Analysis

项目1 小鼠模型分析

• 批准号: 10729466
• 负责人: EDGAR G. ENGLEMAN
• 金额: $ 56.36万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-19 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10729466
• 关键词: Adaptive Immune System; Algorithms; Anatomy; Architecture; Atypical lymphocyte; Automobile Driving; Back; Biological Markers; Blocking Antibodies; Cell Communication; Cells; Chronology; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Computing Methodologies; Cytometry; Data; Development; Diagnosis; Disease; Disease Progression; Distal; Distant; Distant Metastasis; Education; Flow Cytometry; Gene Expression Profile; Genes; Genetic Transcription; Head and Neck Cancer; Human; Image; Immune; Immune Evasion; Immune Tolerance; Kinetics; Knockout Mice; Label; Leukocyte Trafficking; Leukocytes; Lymph Node Involvement; Lymphatic; Malignant Neoplasms; Mediating; Metastatic Neoplasm to Lymph Nodes; Modeling; Mus; Nature; Neighborhoods; Neoplasm Metastasis; Nodal; Non-Small-Cell Lung Carcinoma; Patients; Population; Predisposition; Prevention; Primary Neoplasm; Process; Property; Regulatory T-Lymphocyte; Role; Site; Solid; Stromal Cells; Study models; Systems Biology; Tissues; Tumor Biology; Tumor Promotion; Tumor-infiltrating immune cells; cancer cell; conditioning; cytokine; data integration; high dimensionality; imaging platform; implantation; lymph nodes; markov model; migration; mouse model; neoplastic cell; new therapeutic target; novel; prevent; programs; single-cell RNA sequencing; therapy design; trafficking; transcriptomics; tumor

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.

总结/摘要：项目1 对于大多数实体恶性肿瘤，淋巴结（LN）转移先于进一步扩散，并导致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， 明白我们在小鼠中发现，在定植淋巴结时，肿瘤细胞激活转录程序， 使其能够通过与LN白细胞的相互作用诱导肿瘤特异性免疫耐受。这些 白细胞随后在整个宿主中再循环，导致全身性免疫耐受， 远处部位的转移性种植。重要的是，我们在LN中发现了相同的保守转录特征， 转移，这表明我们的LN转移和免疫耐受的驱动机制 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， LN定植前、定植中和定植后LN内的白细胞和基质改变远处组织， 疾病进展。我们预计这些相互作用涉及同型和异型细胞-细胞相互作用， 协调LN、肿瘤和远端部位内的结构变化，以及各种人群的运输 整个主机。我们将使用同系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进展。1：确定肿瘤发生的动力学 产生系统性免疫耐受。使用高维成像平台（CODEX），我们将揭示 整个宿主局部微环境的纵向变化。第二章：质疑异型细胞-细胞的本质 LN内的相互作用以及这些相互作用促进免疫逃避和诱导的机制 免疫耐受性。我们将使用scRNA-seq来鉴定免疫和基质细胞中的转录变化， 与淋巴结内的恶性细胞相互作用的群体。使用空间转录组学，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处的细胞不同。确认 这些目标和项目2中确定的目标的功能意义，我们将结合联合收割机CRISPR- LN转移性肿瘤的介导的基因编辑，沿着细胞耗竭研究和敲除小鼠或诱导型 小鼠模型。3：确定来自肿瘤涉及的LN的免疫细胞的运输如何建立促转移性淋巴结转移。 微环境在遥远的地方我们将采用一种新的细胞标记平台来评估细胞的贩运。 白细胞和基质细胞从涉及肿瘤的淋巴结转移到远处并返回原发肿瘤。这些 研究将揭示LN转移促进肿瘤向远处扩散的机制， 用于预防和治疗转移性疾病的新靶点。