核骨架蛋白介导MKL1调控实验性近视巩膜重塑的分子机制研究

巩膜重塑、弹性承载力降低是近视眼轴延长的重要环节，然其机制不甚清晰。本课题组前期研究发现，力学敏感分子核骨架蛋白LaminA/C、Emerin在形觉剥夺性近视小鼠巩膜中下调，伴核内MKL1减少，但作用不明。我们推测核骨架蛋白是启动MKL1调节胶原相关基因表达引起巩膜重塑的可能“枢纽”。为此，我们拟对近视动物模型和软性硬性基质培养的细胞模型，运用基因敲除及重组技术，首先研究上调、下调LaminA/C、Emerin是否通过MKL1影响下游的胶原代谢相关因子；其次双向调节MKL1，观察SRF、Sp1及p300、H3K9ac的变化，运用ChIP-seq、RNA-seq及定量PCR检测下游基因的转录及表达；进而分别以Emerin、MKL1为靶标进行在体干预，揭示其可能是近视治疗的有效靶点。迄今这些研究尚属空白，其预期结果将明确近视巩膜重塑中核骨架蛋白相关的力学生物学机制，具有重要的理论和现实意义。

Scleral remodeling and reduced elasticity are important in myopia axial elongation, but the mechanism is not very clear. Our research team found that the mechanically sensitive molecular nuclear envelope proteins LaminA/C and Emerin were significantly down-regulated, as well as MKL1 in the nucleus, in the sclera of form-deprivation myopia mice, but the role was unknown. We speculate that nuclear envelope proteins are "hub" that triggers MKL1 to regulate collagen-related gene expression and cause scleral remodeling. Therefore, we apply gene knockout and recombination technology on myopia animal models and cell models cultured on soft and stiff matrix to explore the mechanism. First, we will study whether up-regulation and down-regulation of LaminA/C and Emerin have effect on downstream collagen metabolism-related factors through MKL1. Secondly, we measure the changes of SRF, Sp1, p300, H3K9ac and use ChIP-seq, RNA-seq and quantitative PCR to detect the transcription and expression changes by modulating MKL1 expression. Furthermore, we target Emerin and MKL1 for intervention, to reveal if it may be a potential therapeutic target in myopia control. To the best of our knowledge, the related research has not been published by now. Our study may clarify the molecular mechanisms related to nuclear envelope proteins in myopic scleral remodeling.