Lymph node regulation of metastatic fitness and reemergence

转移适应性和复发的淋巴结调节

基本信息

批准号：
10593135
负责人：
Andrea Rachel Lim
金额：
--
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2023-04-02
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10593135
关键词：
Acute Affect Affinity Chromatography Aftercare Animals Antibodies Bar Codes Blood Blood Circulation Blood Vessels Bone Marrow Breast Cancer Cell Breast Cancer Model Breast Cancer Patient Breast cancer metastasis Cancer Cell Growth Cancer Etiology Cell Cycle Cessation of life Clinical Clonality Coculture Techniques Color Competence Complex Cues Dangerousness Deposition Disease Endothelium Excision Exposure to Extracellular Matrix Generations Goals Grant Heparan Sulfate Proteoglycan Human Immune Immune Tolerance Immunity In Situ Inflammation Inguinal lymph node group Injections Intervention Lipopolysaccharides Literature Lung Lymph Lymphatic Lymphatic Endothelial Cells Lymphatic Endothelium Malignant Neoplasms Mediating Messenger RNA Metastatic breast cancer Modeling Molecular Monoclonal Antibodies Mus Neoplasm Metastasis Organ Outcome Ovalbumin Pathway interactions Patients Peripheral Phase Population Postdoctoral Fellow Primary Neoplasm Proteomics Regulation Relapse Ribosomes Route Shapes Site System T cell receptor repertoire sequencing T-Lymphocyte T-cell receptor repertoire TNFRSF5 gene Tail Testing Therapeutic Time Tissues Transgenic Mice Translating Tumor Antigens Tumor Promotion Tumor-infiltrating immune cells Veins Work antigen-specific T cells candidate identification candidate validation cell growth differential expression experience fitness functional outcomes immunoregulation in vivo in vivo Model in vivo evaluation insight intravital imaging knock-down lymph nodes malignant breast neoplasm mouse model neoplastic cell novel novel therapeutic intervention overexpression peripheral tolerance permissiveness prevent receptor expression small hairpin RNA targeted treatment tumor

项目摘要

PROJECT SUMMARY Metastasis is the most dangerous stage of cancer, responsible for the vast majority of breast cancer-related deaths. Currently, we have no treatments that target metastasis; therefore, it is critical to understand metastatic progression at the molecular, cellular, and tissue levels to develop therapeutic strategies against this disease. This proposal investigates how the lymph node (LN) regulates tumor dormancy, a stage of quiescence responsible for delayed relapse which affects ~20% of metastatic breast cancer patients. Using this latency window as an opportunity for intervention requires an understanding of site-specific dormancy regulation. Although the LN is the most common site of metastasis, very little is known about how it sustains dormant DTCs and or what triggers reawakening. I hypothesize that stable LN endothelia deposit quiescence factors which are lost when the stable niche is disturbed during inflammation. This proposal combines animal studies, novel models of the LN microvascular niche (MVN), and intravital imaging to identify LN molecules that contribute to DTC quiescence and reawakening. ECM proteomics identified candidate quiescent factors including HSPG4, LOXL1, and TINAGL1 which will be validated in the LN MVNs. Candidates will be functionally tested in the LN MVNs and in vivo. Further, acute inflammation will be used to trigger dormant DTC reawakening in mouse models and by intravital imaging to see whether DTCs “wake up” when their niche is activated. These studies will provide the first mechanistic understanding of how the LN promotes tumor dormancy and demonstrate whether destabilizing the niche through inflammation is sufficient to wake dormant tumor cells up. In the K00 phase, I propose to investigate route of dissemination as a selective force that enriches metastasis-competent populations of tumor cells. Lymphatics are not only less hostile than blood circulation, but present an opportunity for immune escape as LN LEC induce peripheral tolerance in the T cell repertoire. LN LEC-mediated deletion of tumor-specific T cells27 may grant a survival advantage to lymph- exposed DTCs, priming them for immune escape. Metastatic advantage of lymphatic dissemination will be investigated using lymphatic-Cre transgenic mouse models to track dissemination route and metastatic outcome, T cell receptor sequencing to elucidate the systemic effect of local deletion on the peripheral repertoire, and molecular barcoding to track clonality and distribution of DTCs relative to their dissemination route. Understanding how dissemination influences metastasis will help identify which tumor cells are most likely to metastasize for elimination. The proposed aims will shed light on how the LN and lymphatic dissemination instruct metastasis, revealing actionable targets against tumor dormancy and metastatic fitness, provide invaluable experience studying the complex interactions between metastasis and immunity.

项目概要转移是癌症最危险的阶段，是绝大多数乳腺癌相关疾病的罪魁祸首目前，我们没有针对转移的治疗方法；因此，了解这一点至关重要。分子、细胞和组织水平的转移进展，以制定针对其的治疗策略该提案研究了淋巴结 (LN) 如何调节肿瘤休眠（肿瘤休眠的一个阶段）。静止期导致延迟复发，影响约 20% 的转移性乳腺癌患者。这种延迟窗口作为干预的机会需要了解特定位点的休眠尽管淋巴结是最常见的转移部位，但人们对其如何维持知之甚少。休眠的 DTC 和/或触发重新唤醒的因素我采取了稳定的 LN 内皮沉积物。当炎症期间稳定的生态位受到干扰时，静止因子就会丢失。结合动物研究、LN 微血管生态位 (MVN) 的新模型和活体成像来识别有助于 DTC 静止和重新唤醒的 LN 分子通过 ECM 蛋白质组学鉴定出候选分子。静态因子包括 HSPG4、LOXL1 和 TINAGL1，将在 LN MVN 中进行验证。将在 LN MVN 和体内进行功能测试此外，将使用急性炎症来触发。在小鼠模型中休眠的 DTC 重新唤醒，并通过活体成像观察 DTC 是否在以下情况下“苏醒”：他们的利基被激活。这些研究将提供对闪电网络如何促进的第一个机制理解。肿瘤休眠并证明通过炎症破坏生态位稳定是否足以唤醒在 K00 阶段，我建议研究作为选择性力量的传播途径。丰富的具有转移能力的肿瘤细胞群不仅比血液更不具有敌意。循环，但由于 LN LEC 诱导 T 细胞外周耐受，因此提供了免疫逃逸的机会 LN LEC 介导的肿瘤特异性 T 细胞删除27 可能会赋予淋巴癌生存优势。暴露的 DTC，启动它们的免疫逃逸将是淋巴传播的转移优势。使用淋巴-Cre 转基因小鼠模型进行研究以追踪传播途径和转移结果，T细胞受体测序阐明局部缺失对外周的系统影响库和分子条形码，用于跟踪 DTC 相对于其传播的克隆性和分布了解传播如何影响转移将有助于确定哪些肿瘤细胞最常见。拟议的目标将阐明淋巴结和淋巴管如何转移。传播指导转移，揭示针对肿瘤休眠和转移适应性的可行靶点，提供研究转移与免疫之间复杂相互作用的宝贵经验。