从S1P/S1PRs调控胶质细胞激活探讨青光眼视神经损伤的机制和治疗

青光眼是不可逆性神经致盲性眼病，目前缺乏有效靶向治疗方法。本课题组已证实，视网膜小胶质细胞激活，通过炎症小体信号通路介导RGCs损伤，但在多种胶质细胞中如何启动炎症小体介导的免疫反应仍未清楚。通过前期单细胞测序及预实验发现：急性高眼压作用下，S1P/S1PRs在视网膜小胶质细胞和星形胶质细胞中表达显著升高，靶向S1PRs可以调控胶质细胞激活，恢复炎症小体平衡，有效保护RGCs存活和视神经功能，但分子机制尚待阐明。因此我们提出假说，S1P/S1PRs启动胶质细胞中炎症小体介导的免疫反应，诱导青光眼视神经损伤。为了验证这一假说，本研究拟利用单细胞测序及条件性基因敲除鼠，探讨胶质细胞激活的时空变化及其分子机制，并通过靶向S1P/S1PRs关键信号通路，探讨胶质细胞介导的免疫炎症反应在青光眼视神经损伤及保护的作用机制，为青光眼的精准治疗提供新的靶点，为视神经保护和轴突再生临床转化提供新思路与途径。

Glaucoma, as a degenerative optic neuropathy, is the leading cause of irreversible blindness. However, an effective targeting treatment is currently lacking for it. We have confirmed that the retinal ganglion cells dysfunction and loss were mediated by the pyroptosis/inflammasomes following microglia activation. Through Single-cell RNA sequencing (scRNA-seq) and preliminary experiments, we found that the level of S1P/S1PRs was significantly elevated in astrocytes and microglia following IOP elevation. Targeting S1PRs could regulate inflammasome-related immune responses in glia and protect the survival of RGCs, axonal regeneration, but the molecular mechanisms remained to be elucidated. Therefore, we propose the hypothesis that the S1P/S1PRs initiates the inflammasome-related immune response to glia activation and induces optic neuropathy in glaucoma. In order to verify this hypothesis, in this study, we used scRNA-seq and conditional gene knockout mice to explore the spatial-temporal change and molecular mechanism of glia activation. And exploring the mechanism of glia-mediated immune responses in glaucoma by targeting S1P/S1PRs key signaling pathway. These results will provide novel therapeutic targets for glaucoma pathogenesis and suggest novel therapeutic approaches for further clinical transformation about optic nerve neuroprotection and axon regeneration.