Project-2:Defining the role of compartmentalized neuro-lymphatic networks on CRC and metastatic progression

项目 2：定义分区神经淋巴网络对 CRC 和转移进展的作用

• 批准号: 10688116
• 负责人: Daniel S Mucida
• 金额: $ 37.48万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-23 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10688116
• 关键词: Address; Affect; Antigens; Antitumor Response; Architecture; Bioinformatics; Biology; Cell Communication; Cell Separation; Cells; Cellular Metabolic Process; Collection; Colorectal Cancer; Communication; Critical Pathways; Cues; Data; Diet; Distal; Drainage procedure; Early identification; Enteral; Environment; Epithelial Cells; Equilibrium; Event; Food Hypersensitivity; Gastrointestinal tract structure; Gene Expression; Genetic; Gnotobiotic; Image; Imaging Techniques; Immune; Immune response; Immune system; Immunologic Surveillance; Immunologics; In Vitro; Indigenous; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Intestinal Neoplasms; Intestines; Invaded; Irritable Bowel Syndrome; Label; Liver; Location; Lymph Node Drainage; Lymphatic; Lymphatic System; Malignant Neoplasms; Maps; Mediating; Metastatic Neoplasm to the Liver; Modernization; Molecular; Mucous Membrane; Neoplasm Metastasis; Neuroimmune; Neurons; Patients; Persons; Physiology; Play; Population; Primary Neoplasm; Resistance; Role; Route; Signal Transduction; Site; Submucosa; System; Tissues; Translating; Tumor Immunity; United States; Visualization; adaptive immunity; cell injury; cell killing; colorectal cancer metastasis; colorectal cancer prevention; colorectal cancer progression; cytokine; draining lymph node; enteric pathogen; gain of function; genetic approach; imaging approach; intestinal villi; loss of function; lymph nodes; lymphatic drainage; lymphatic vessel; metabolomics; metastatic colorectal; microbial; microbiota; mouse model; neuronal cell body; neuronal circuitry; novel; novel strategies; prevent; response; screening; tool; transcriptomics; translatome; tumor; tumor microenvironment; tumor progression

Colorectal cancer (CRC) is the second most deadly cancer in the United States, affecting over 140,000 people each year, killing approximately 50,000 in the US, largely by metastatic progression. The intestine hosts the body’s largest collection of immune cells, maintained in close proximity to foreign antigens from the diet, enriched in the proximal regions, and microbiota, which accumulate in the distal regions. The vast and highly connected gut lymphatic vessels form a major cell- and antigen transport route to the draining lymph nodes, responsible for the initiation of adaptive immunity, which could play four roles in regulating colorectal cancer metastasis: immune cells could (i) prevent CRC progression and early dissemination or metastasis by immune-cell killing (ii) promote CRC progression and metastasis through release of inflammatory cytokines, (iii) prevent metastatic colonization in the liver, or (iv) promote metastatic colonization in the liver. We provide data that compartmentalization of intestinal lymphatic drainage to functionally distinct lymph nodes facilitates the simultaneous induction of immune-suppressive and inflammatory immune responses in the gut; however, the relevance of gut lymphatics to CRC and metastasis remains underexplored. In addition to compartmentalized lymphatic networks, the gut also hosts a large number of enteric neurons functionally tuned to each region they occupy. Using retrograde tracing from distinct intestinal regions and specific cell-sorting independent transcriptomics, we uncovered novel neuronal circuits and a role for enteric neurons in sensing perturbations in the intestinal tissue; whether enteric neurons sense and modulate CRC metastatic progression remains unknown. Overall, Project 2 is focused on understanding how enteric-associated neurons sense and provide signals that regulate CRC progression and liver dissemination and how compartmentalized intestinal lymphatic drainage of primary tumor and metastatic liver sites regulate anti-tumor responses and early dissemination. In Aim 1, we hypothesize that primary CRC, as well as liver metastasis, are specifically sensed by populations of enteric-associated neurons, which in turn, influence tumor and metastatic progression. This question will be addressed using tools to visualize and circuit- map, single cell and active translating transcriptomics, and chemogenetic approaches targeting specific neuronal subsets. In Aim 2, we hypothesize that the intestinal lymphatic system communicates primary CRC and early metastatic seeding to the local and systemic immune system, modulating anti-tumor responses. This will be addressed combining modern clearing and live imaging techniques, single cell transcriptomics and novel immune cell-interaction approaches. By combining these approaches, expertise of Mucida lab, with Sohail Tavazoie lab’s expertise in cancer and metastasis biology (Project 1), the Birsoy lab’s expertise in in vitro screenings and cell metabolism (Project 3), and the Cao and Saeed Tavazoie labs expertise in single-cell and bioinformatics analyses, we seek to determine the role of neuro-lymphatic networks in CRC progression and metastatic formation.

结直肠癌(CRC)是美国第二大致命性癌症，影响超过14万人 每年，在美国大约有50,000人死亡，主要是由于转移进展。肠道承载着 体内最大的免疫细胞集合，与饮食中的外来抗原保持密切联系，富含 在近端区域，以及在远端区域积累的微生物区系。广袤且高度相连的 肠道淋巴管形成一条主要的细胞和抗原运输路线，到达引流的淋巴结节，负责 启动适应性免疫，在调节结直肠癌转移中可能发挥四个作用：免疫 细胞可(I)通过免疫细胞杀伤防止结直肠癌进展和早期扩散或转移(Ii)促进 通过释放炎性细胞因子促进结直肠癌进展和转移；(Iii)防止转移定植 或(Iv)促进肝脏的转移性定植。我们提供的数据表明， 肠淋巴引流到功能不同的淋巴结有助于同时诱导 肠道中的免疫抑制和炎性免疫反应；然而，肠道淋巴管的相关性 对结直肠癌和转移的研究仍然很少。除了分隔的淋巴网络外，肠道 也拥有大量的肠道神经元，功能上调节到它们所在的每个区域。使用逆行 通过追踪不同的肠道区域和特定的细胞分选独立转录组，我们发现了新的 神经回路和肠神经元在感知肠道组织中的扰动中的作用 神经元对结直肠癌转移的感知和调控尚不清楚。总体而言，项目2的重点是 了解肠道相关神经元如何感知和提供信号来调节CRC的进展和 肝脏播散与原发灶和转移瘤肠道淋巴引流的区划 肝脏部位调节抗肿瘤反应和早期扩散。在目标1中，我们假设主CRC， 以及肝脏转移，都是由肠道相关神经元群体特异性地感觉到的，这些神经元反过来， 影响肿瘤和转移进展。这个问题将使用工具来可视化和巡回- MAP、单细胞和主动翻译转录，以及针对特定神经元的化学发生方法 子集。在目标2中，我们假设肠道淋巴系统与原发的结直肠癌和早期 转移播撒到局部和全身免疫系统，调节抗肿瘤反应。这将是 结合现代透明和实时成像技术、单细胞转录和新型免疫 细胞相互作用的方法。通过将这些方法结合起来，Mucida实验室的专业知识与Sohail Tavazoie实验室的 在癌症和转移生物学方面的专业知识(项目1)，Birsoy实验室在体外筛选和细胞方面的专业知识 代谢(项目3)，以及CaO和Saeed Tavazoie实验室在单细胞和生物信息学方面的专业知识 分析，我们试图确定神经淋巴网络在结直肠癌进展和转移中的作用 队形。