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: 10493342
• 负责人: Daniel S Mucida
• 金额: $ 31.69万
• 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/10493342
• 关键词: Address; Affect; Antigens; Antitumor Response; Architecture; Bioinformatics; Biology; Cell Communication; Cell Separation; Cells; Cellular Metabolic Process; Collection; Colorectal Cancer; Critical Pathways; Cues; Data; Diet; Distal; Drainage procedure; Enteral; Environment; Epithelial Cells; Equilibrium; Event; Food Hypersensitivity; Gastrointestinal tract structure; Gene Expression; Gnotobiotic; Image; Imaging Techniques; Immune; Immune response; Immune system; Immunologic Surveillance; Immunologics; In Vitro; Indigenous; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Intestinal Neoplasms; Intestines; Irritable Bowel Syndrome; Label; Lead; 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; System; Tissues; Translating; Tumor Immunity; United States; adaptive immunity; cell injury; cell killing; colorectal cancer metastasis; colorectal cancer prevention; colorectal cancer progression; cytokine; draining lymph node; enteric pathogen; 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) 是美国第二大致命癌症，影响超过 140,000 人 每年，美国约有 50,000 人死亡，主要是由于转移性进展。肠道承载着 身体最大的免疫细胞集合，与饮食中的外来抗原保持密切接触，丰富了 近端区域的微生物群和远端区域积累的微生物群。幅员辽阔且联系紧密 肠道淋巴管形成了通往引流淋巴结的主要细胞和抗原运输路线，负责 适应性免疫的启动，在调节结直肠癌转移中发挥四个作用： 细胞可以 (i) 通过免疫细胞杀伤来预防 CRC 进展和早期传播或转移 (ii) 促进 通过释放炎症细胞因子促进结直肠癌进展和转移，(iii) 防止转移定植 在肝脏中，或（iv）促进肝脏中的转移定植。我们提供划分的数据 肠道淋巴引流到功能不同的淋巴结有利于同时诱导 肠道内的免疫抑制和炎症免疫反应；然而，肠道淋巴管的相关性 CRC 和转移的影响仍未得到充分研究。除了分区的淋巴网络外，肠道 还拥有大量肠神经元，这些神经元在功能上与它们占据的每个区域相适应。使用逆行 从不同的肠道区域和特定的细胞分选独立转录组学追踪，我们发现了新的 神经元回路以及肠神经元在感知肠道组织扰动中的作用；是否肠溶 神经元感知和调节 CRC 转移进展仍然未知。总体而言，项目 2 的重点是 了解肠相关神经元如何感知和提供调节 CRC 进展的信号，以及 肝脏播散以及如何区分原发性肿瘤和转移性肿瘤的肠道淋巴引流 肝脏部位调节抗肿瘤反应和早期传播。在目标 1 中，我们假设原发性 CRC， 以及肝转移，被肠道相关神经元群体特异性感知，反过来， 影响肿瘤和转移进展。这个问题将使用可视化和电路工具来解决 图谱、单细胞和主动翻译转录组学以及针对特定神经元的化学遗传学方法 子集。在目标 2 中，我们假设肠道淋巴系统沟通原发性 CRC 和早期 转移播种到局部和全身免疫系统，调节抗肿瘤反应。这将是 解决了现代透明化和实时成像技术、单细胞转录组学和新型免疫相结合的问题 细胞相互作用方法。通过将这些方法、Mucida 实验室的专业知识与 Sohail Tavazoie 实验室的 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 新陈代谢（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 通过分析，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 形成。