Song Yifan, Xu Zhe, Li Hong-Tao, Xie Yunxiao, Zhao Lianghui, Feng Jiaojiao, Luo Anfeng, Dai Jiajing, Li Jing, Guo Xinran, Song Jike, Bi Hongsheng
Shandong University of Traditional Chinese Medicine, Jinan, P. R. China.
Institute of Brain Science and Brain-Inspired Research, Shandong First Medical University & Shandong Academy of Medical Sciences, Jinan, P. R. China; Shandong Institute of Brain Science and Brain-Inspired Research, Jinan, China.
Mol Cell Proteomics. 2025 Nov;24(11):101069. doi: 10.1016/j.mcpro.2025.101069. Epub 2025 Sep 19.
Myopia is a growing global public health concern. Recent studies have revealed that the regulation of eye growth occurs via a complex signaling cascade, which originates in the retina and across the choroid to the sclera. Identifying key proteins and specific biological processes (BPs) in the retina, choroid, and sclera is crucial for understanding the molecular mechanisms underlying myopia development. We conducted comprehensive proteomic and phosphoproteomic analyses of the retina, choroid, and sclera from form-deprivation myopia guinea pigs using liquid chromatography-tandem mass spectrometry. Differentially expressed proteins and phosphosites were identified, followed by functional annotation and signaling pathway enrichment analyses. The expression of key proteins was assessed using Western blotting and enzyme-linked immunosorbent assay. Distinct proteomic and phosphoproteomic profiles were observed across the three tissues, with 6470, 6708, and 3236 proteins and 9613, 9416, and 3685 phosphosites in the retina, choroid, and sclera, respectively. Proteomic analysis showed that neural signal transduction was enriched in the retina, with downregulation of NTRK2, suggesting impaired neurotrophic signaling. The upregulation of SYK and BTK, along with increased NF-κB, p65, and IL-1β levels in the choroid, indicated enhanced inflammatory responses. TNNT3, TPM2, and ACTN3 were upregulated in the sclera, reflecting cytoskeletal remodeling associated with scleral expansion. Phosphoproteomic analysis indicated key roles of phosphoproteins in BPs, particularly the spliceosome signaling pathway, which was broadly involved across all three tissues. Kinase network analysis revealed PRPF4B as a key kinase for SF3B1, suggesting the potential regulation roles of RNA splicing in myopia progression. The present study systematically elucidates the proteomic and phosphoproteomic characteristics of the retina, choroid, and sclera of form-deprivation myopia in guinea pigs, highlighting significant tissue-specific BPs to myopia. The findings provide a theoretical foundation for understanding that different tissues exhibit distinct biological reactions to myopia, each through specific signaling pathways and regulatory mechanisms.
近视是一个日益严重的全球公共卫生问题。最近的研究表明，眼球生长的调节是通过一个复杂的信号级联反应发生的，该反应起源于视网膜，穿过脉络膜到达巩膜。识别视网膜、脉络膜和巩膜中的关键蛋白质和特定生物学过程（BP）对于理解近视发展的分子机制至关重要。我们使用液相色谱-串联质谱对形觉剥夺性近视豚鼠的视网膜、脉络膜和巩膜进行了全面的蛋白质组学和磷酸化蛋白质组学分析。鉴定了差异表达的蛋白质和磷酸化位点，随后进行了功能注释和信号通路富集分析。使用蛋白质免疫印迹法和酶联免疫吸附测定法评估关键蛋白质的表达。在这三种组织中观察到了不同的蛋白质组学和磷酸化蛋白质组学谱，视网膜、脉络膜和巩膜中分别有6470、6708和3236种蛋白质以及9613、9416和3685个磷酸化位点。蛋白质组学分析表明，神经信号转导在视网膜中富集，NTRK2下调，表明神经营养信号受损。脉络膜中SYK和BTK的上调以及NF-κB、p65和IL-1β水平的增加表明炎症反应增强。TNNT3、TPM2和ACTN3在巩膜中上调，反映了与巩膜扩张相关的细胞骨架重塑。磷酸化蛋白质组学分析表明磷酸化蛋白质在生物学过程中起关键作用，特别是剪接体信号通路，该通路广泛涉及所有三种组织。激酶网络分析揭示PRPF4B是SF3B1的关键激酶，表明RNA剪接在近视进展中可能具有调节作用。本研究系统地阐明了形觉剥夺性近视豚鼠视网膜、脉络膜和巩膜的蛋白质组学和磷酸化蛋白质组学特征，突出了对近视具有显著组织特异性的生物学过程。这些发现为理解不同组织通过特定信号通路和调节机制对近视表现出不同的生物学反应提供了理论基础。
