Role of meningeal lymphatic vasculature in neuroimmune communication development

该提案的目标是首次确定脑膜是如何形成的 淋巴管(MLV)在出生后早期影响神经免疫通讯，如果MLV- 幼鼠依赖的神经免疫通讯可被血管内皮生长因子-C信号所控制 在稳定状态下，在儿童脑肿瘤模型中，如髓母细胞瘤，最 影响儿童的常见肿瘤。 我们以前的工作和初步数据表明，MLV在出生后早期发育， 它们的生长需要血管内皮生长因子-C(VEGF-C)和血管内皮生长因子-C(VEGF-C)共同作用。 受体Neuropilin2(Nrp2)，促进MLV生长使适应性免疫 成人对胶质母细胞瘤的反应。这些发现促使我们研究 MLV介导的神经免疫连接是如何建立的，并表征了 中枢神经系统集合淋巴结的抗原提呈细胞和MLV依赖 T细胞在发育中的培养。 成功完成这一项目的实验方法包括新的淋巴特异性 利用光片进行三维全头成像的新方法--敲击线 荧光显微镜，以及质谱仪和单细胞的无偏分子分析- RNA测序。 这项多学科研究将汇集以下领域的互补专业知识 脑膜淋巴管(Jean-Leon Thomas，耶鲁)，发育中的血管生成(Anne Eichmann， 耶鲁大学)和免疫学(岩崎明子，耶鲁大学)，以及其他儿科肿瘤学顾问(Asher 耶鲁大学儿科神经肿瘤学主任)，液-色质谱计 (耶鲁大学Keck MS和蛋白质组学资源主任Tukiet Lam)和TCR图谱分析 (张乐，耶鲁神经病学)。 我们希望描述MLV依赖的神经免疫通讯的发展， 确定Nrp2为MLV发芽的新的强制性调控因子，确定APC亚群(S) CNS抗原转运到cLN所需的，并确定作为原则证据的 血管内皮生长因子-C驱动的MLV生长促进针对CNS异常的早期T细胞教育的潜力 抗原，具有治疗儿童侵袭性脑肿瘤的前景。

The objectives of the proposal are to determine, for the first time, how developing meningeal lymphatic vessels (MLVs) impact neuroimmune communication in early postnatal life, and if MLV- dependent neuroimmune communication can be harnessed by VEGF-C signaling in young mice at steady state and in a model of pediatric brain tumor such as medulloblastoma, the most common tumor affecting children. Our previous work and preliminary data show that MLVs develop during early postnatal life, that their growth requires Vascular Endothelial Growth Factor-C (VEGF-C) and the VEGF-C co- receptor Neuropilin2 (Nrp2), and that enhancing MLV growth enables adaptive immune responses against glioblastoma in the adult. These findings prompt us to investigate mechanims how MLV-mediated neuroimmune connection establishes and to characterize the transport of antigen-presenting cells (APCs) to CNS-collecting lymph nodes (cLNs) and the MLV-dependent education of T cells in developing cLNs. Experimental approaches to successfully complete this project include new lymphatic-specific knockout lines, new procedures of three-dimensional whole-head imaging by light sheet fluorescence microscopy, and unbiased molecular analysis by mass spectrometry and single cell- RNA sequencing. This multidisciplinary study will bring together complementary expertise in the fields of meningeal lymphatics (Jean-Leon Thomas, Yale), developmental angiogenesis (Anne Eichmann, Yale), and immunology (Akiko Iwasaki, Yale), with additional advisors in pediatric oncology (Asher Marks, Director of Yale Pediatric Neuro-Oncology), liquid-chromatography mass spectometry (Tukiet Lam, Director of Keck MS & Proteomics Resource at Yale), and TCR repertoire analysis (Le Zhang, Yale Neurology). We expect to characterize the development of MLV-dependent neuroimmune communication, to identify Nrp2 as a new mandatory regulator of MLV sprouting, identify APC subpopulation(s) required for CNS antigen transport into cLNs, and to determine, as a proof of principle, the potential of VEGF-C-driven MLV growth to promote early T cell education against abnormal CNS antigens, with perspectives for the treatment of aggressive pediatric brain tumors.