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.

摘要/摘要：项目 1 大多数实体恶性肿瘤在进一步扩散之前发生淋巴结 (LN) 转移，并进入 III 期 诊断。尽管如此，淋巴结转移在疾病进一步进展中的机制作用尚不清楚。 明白了。我们在小鼠身上发现，在定殖淋巴结时，肿瘤细胞激活转录程序， 通过与淋巴结白细胞的相互作用，能够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，从而促进全身免疫耐受 远处部位的转移播种。重要的是，我们在 LN 中发现了相同的保守转录特征 人类转移，表明驱动淋巴结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 内的白细胞和基质会改变远处的组织，以促进 疾病进展。我们期望这些相互作用涉及同型和异型细胞间相互作用， 协调 LN、肿瘤和远处站点内的架构变化以及不同群体的贩运 整个主机。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进展。 1：确定肿瘤的动力学 产生全身免疫耐受。使用高维成像平台（CODEX），我们将发现 宿主局部微环境的纵向变化。 2：探究异型细胞间的本质 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 的免疫耐受性。我们将使用 scRNA-seq 来识别免疫和基质中的转录变化 群体与 LN 内的恶性细胞相互作用。使用空间转录组学，我们将确定如何 与恶性细胞相互作用的细胞的基因表达模式不同于远处的细胞。为了确认 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- 介导的 LN 转移性肿瘤基因编辑，以及细胞耗竭研究和基因敲除小鼠或诱导型小鼠 鼠标模型。 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转移 遥远地点的微环境。我们将采用一种新颖的细胞标记平台来评估 白细胞和基质细胞从肿瘤相关的淋巴结转移到远处，然后返回原发肿瘤。这些 研究将揭示淋巴结转移促进肿瘤扩散到远处部位的机制，并确定 预防和治疗转移性疾病的新靶点。