Lymph node regulation of metastatic fitness and reemergence

转移适应性和复发的淋巴结调节

• 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
• 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

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.

项目摘要 转移是癌症最危险的阶段，负责绝大多数与乳腺癌有关的阶段 死亡人数。目前，我们没有针对转移的治疗方法。因此，了解 分子，细胞和组织水平的转移进展，以制定反对的理论策略 这种疾病。该提案调查了淋巴结（LN）如何调节肿瘤休眠 导致延迟继电器的静止会影响约20％的转移性乳腺癌患者。使用 这个延迟窗口作为干预的机会，需要了解特定地点的休眠 规定。尽管LN是转移的最常见位置，但对其如何维持很少了解 休眠的DTC和或触发的内容正在唤醒。我假设稳定的LN内皮矿床 稳定的小众在炎症期间被干扰时丢失的静止因子。这个建议 结合动物研究，LN微血管小裂（MVN）的新型模型和浸润成像以鉴定 有助于DTC静止和唤醒的LN分子。 ECM蛋白质组学确定了候选者 将在LN MVN中验证的HSPG4，LOXL1和TINAGL1在内的静止因子。候选人 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 休眠的DTC在小鼠模型中重新唤醒，并通过插入式成像来查看DTC是否在 他们的利基被激活。这些研究将提供对LN如何促进的第一个机械理解 肿瘤休眠并证明是否通过注射破坏利基市场足以唤醒 休眠的肿瘤细胞向上。在K00阶段，我建议研究传播作为选择性力的途径 这丰富了肿瘤细胞的转移群体。淋巴管不仅比血液遭受敌意 循环，但由于LN LEC在T细胞中诱导外围耐受性，因此提供了免疫逃逸的机会 曲目。 LN LEC介导的肿瘤特异性T细胞的缺失27可能给予淋巴的生存优势 暴露的DTC，将它们启动以进行免疫逃生。淋巴传播的转移优势将是 使用淋巴细胞转基因小鼠模型进行了研究以跟踪传播途径和转移 结果，T细胞受体测序以阐明局部缺失对周围的全身效应 曲目和分子条形码以跟踪DTC相对于其传播的克隆性和分布 路线。了解传播如何影响转移将有助于确定哪些肿瘤细胞最多 可能转移以消除。拟议的目标将阐明LN和淋巴样 传播指导转移，揭示了针对肿瘤休眠和转移性适应性的可行靶标， 提供研究转移与免疫之间复杂相互作用的宝贵经验。