UTX调节血管免疫沟通机制及在急性脊髓损伤中的作用

Acute Spinal Cord Injury (SCI) is a serious central nervous system traumatic disease. The vascular immune response induced by the injury of spinal cord microvascular endothelial cells (SCMECs) after SCI is the main reason to aggravate SCI. Previously, we found that UTX, an epigenetic regulatory factor, can regulate the expression of multiple regeneration-related genes in SCMECs, of which CTGF is the most significant. It further affects the polarization of macrophages and the repair of neurological function after SCI. However, the SCMECs regeneration regulated by UTX, the vascular immune response induced by the epigenetic regulation of SCMECs, the communication of SCMECs to macrophages and the related mechanisms are still unknown. In this project, the epigenetic mechanism by which UTX regulates SCMECs regeneration and its effect on the macrophages polarization are going to be explored based on the co-culture system of primary SCMECs and macrophages, the endothelial specific UTX knockout mice model, the advanced synchrotron radiation 3D imaging and the molecular biology techniques; the macrophage phagocytized exosomes released by SCMECs and its effect on the PI3K/Akt/mTOR signaling pathway of macrophages will be detected by the extraction of exosomes, the confocal live cell imaging and in vivo tracing system. Hence, the mechanism of vascular communication to immune and its role in the repair of spinal cord injury can be elucidated. This project can explore the new target for the treatment of SCI.

急性脊髓损伤（SCI）是一种严重的神经系统创伤性疾病。SCI后脊髓微血管内皮细胞（SCMECs）损伤诱发血管免疫炎症反应是加重脊髓损伤的主要原因。课题组前期研究发现表观调节因子UTX可调控SCMECs多个血管再生相关基因的表达，其中以CTGF最为显著，进而影响巨噬细胞极化和神经功能修复。然而UTX靶向调节SCMECs再生、血管免疫炎症反应及SCMECs与巨噬细胞沟通机制不明。本项目拟建立原代SCMECs/巨噬细胞共培养体系，结合血管内皮细胞条件性UTX敲除动物模型，利用同步辐射3D成像和分子生物学技术，揭示UTX表观调节SCMECs再生机制以及对巨噬细胞极化的影响；同时提取SCMECs外泌体，采用共聚焦活体细胞成像和体内示踪检测外泌体与巨噬细胞结合，及其对巨噬细胞PI3K/Akt/mTOR信号通路活化的影响，从而阐明血管与免疫沟通机制及其在脊髓损伤修复中的作用，为治疗SCI提供新靶点。

急性脊髓损伤（SCI）是一种严重的神经系统创伤性疾病。SCI后脊髓微血管内皮细胞（SCMECs）损伤诱发血管免疫炎症反应是导致继发性损伤的主要原因。本项目“UTX调节血管免疫沟通机制及在急性脊髓损伤中的作用”通过建立内皮/巨噬细胞共培养体系，结合血管内皮细胞条件性UTX敲除动物模型，利用同步辐射3D成像、共聚焦活体细胞成像、外泌体体内/外示踪等技术揭示了脊髓损伤后表观因子UTX通过调控SCMECs再生影响巨噬细胞极化过程，并进一步验证了内皮细胞外泌体调控巨噬细胞活化的下游信号通路，从而阐明血管与免疫沟通机制及其在脊髓损伤修复中的作用，为治疗SCI提供新靶点。.本研究计划主要涉及以下3个方面：1）明确脊髓微血管内皮细胞条件性UTX敲除（SCMECs UTX-cKO）对SCI后SCMECs再生和巨噬细胞极化及小鼠神经功能恢复的影响；2）探究UTX靶向调节SCMECs的CTGF表达对血管免疫和SCI后神经功能恢复的影响；3）验证UTX表观调节SCMECs外泌体CTGF介导血管与免疫细胞沟通机制及在SCI中的作用。本研究从结构-功能两方面系统阐释UTX 靶向调节脊髓微血管内细胞CTGF 的表达对SCI 后血管再生和免疫的调节作用；并从分子、细胞及整体水平深入阐释UTX-CTGF 调节SCMECs外泌体介导血管与免疫细胞沟通机制，进一步明确其在SCI 过程中的作用，为其促进脊髓神经功能恢复、治疗SCI 提供实验基础与理论依据。综上，结合研究计划实施、调整以及扩展情况，本项目顺利完成了既定目标。