Han Fang, Wang Leyi, Wu Jiayin, Shen Lin, Li Yangyang, Guo Hui, Li Jianqiao
Department of Ophthalmology, Qilu Hospital of Shandong University, Jinan, China.
The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Health, The State and Shandong Province Joint Key Laboratory of Translational Cardiovascular Medicine, Qilu Hospital of Shandong University, Jinan, China.
Invest Ophthalmol Vis Sci. 2025 Feb 3;66(2):13. doi: 10.1167/iovs.66.2.13.
The purpose of this study was to investigate the role of LRRK2 in the inflammatory response to fungal keratitis (FK) and elucidate the underlying mechanisms.
The protein levels of leucine-rich repeat kinase 2 (LRRK2), p-LRRK2, and stimulator of interferon genes (STING)-related proteins were assessed by western blot analysis. ELISA and quantitative real-time polymerase chain reaction (qRT-PCR) were employed to evaluate the inflammatory response induced by Aspergillus fumigatus. Mass spectrometry was performed to identify the interaction partners of LRRK2. The glutathione S-transferase (GST) pull-down assay and co-immunoprecipitation (co-IP) were used to verify the interaction between LRRK2 and STING. Additionally, fungal load determinations and clinical score assessments were conducted to determine corneal infection in a mouse model.
A. fumigatus stimulation promoted the phosphorylation of LRRK2 through Toll-like receptor 2 (TLR2) in human corneal epithelial cells (HCECs) and mouse corneas. LRRK2 overexpression enhanced the A. fumigatus-induced inflammatory response, and LRRK2 knockdown alleviated A. fumigatus keratitis both in vitro and in vivo. Mass spectrometry identified STING as a novel interaction partner of LRRK2. Moreover, A. fumigatus treatment enhanced the interaction between LRRK2 and STING, resulting in the phosphorylation and activation of STING. The phosphorylated STING then triggered its downstream signaling pathways, exacerbating the severity of A. fumigatus keratitis. LRRK2 inhibitor (LRRK2-IN-1) significantly mitigated the inflammatory response and corneal damage caused by A. fumigatus stimulation.
LRRK2 inhibition ameliorates A. fumigatus-induced inflammation through modulating STING signaling pathways in both HCECs and mouse models. Our results suggest that targeted inhibition of LRRK2 could be a promising strategy for FK treatment.
本研究旨在探讨富含亮氨酸重复激酶2（LRRK2）在真菌性角膜炎（FK）炎症反应中的作用，并阐明其潜在机制。
采用蛋白质印迹分析评估富含亮氨酸重复激酶2（LRRK2）、磷酸化LRRK2和干扰素基因刺激因子（STING）相关蛋白的水平。采用酶联免疫吸附测定（ELISA）和定量实时聚合酶链反应（qRT-PCR）评估烟曲霉诱导的炎症反应。进行质谱分析以鉴定LRRK2的相互作用伙伴。采用谷胱甘肽S-转移酶（GST）下拉试验和免疫共沉淀（co-IP）验证LRRK2与STING之间的相互作用。此外，进行真菌载量测定和临床评分评估以确定小鼠模型中的角膜感染情况。
烟曲霉刺激通过Toll样受体2（TLR2）促进人角膜上皮细胞（HCECs）和小鼠角膜中LRRK2的磷酸化。LRRK2过表达增强了烟曲霉诱导的炎症反应，而LRRK2基因敲低在体外和体内均减轻了烟曲霉角膜炎。质谱分析鉴定出STING是LRRK2的新型相互作用伙伴。此外，烟曲霉处理增强了LRRK2与STING之间的相互作用，导致STING磷酸化并激活。磷酸化的STING随后触发其下游信号通路，加剧了烟曲霉角膜炎的严重程度。LRRK2抑制剂（LRRK2-IN-1）显著减轻了烟曲霉刺激引起的炎症反应和角膜损伤。
在HCECs和小鼠模型中，抑制LRRK2可通过调节STING信号通路改善烟曲霉诱导的炎症。我们的结果表明，靶向抑制LRRK2可能是一种有前景的FK治疗策略。
