Laboratory of Experimental Optometry, Centre for Myopia Research, School of Optometry, The Hong Kong Polytechnic University, Hung Hom, Kowloon, Hong Kong.
Laboratory of Experimental Optometry, Centre for Myopia Research, School of Optometry, The Hong Kong Polytechnic University, Hung Hom, Kowloon, Hong Kong.
J Proteomics. 2020 Jun 15;221:103684. doi: 10.1016/j.jprot.2020.103684. Epub 2020 Feb 13.
Myopia, the most common cause of impaired vision, may induce sight- threatening diseases or ocular complications due to axial elongation. The exact mechanisms underlying myopia development have received much attention and understanding of these is necessary for clinical prevention or therapeutics. In this study, quantitative proteomics using Isotope Coded Protein Label (ICPL) was applied to identify differentially regulated proteins in the retinas of myopic chicks and, from their presence, infer the possible pathogenesis of excessive ocular elongation. Newly hatched white leghorn chicks (n = 15) wore -10D and + 10D lenses bilaterally for 3 and 7 days, respectively, to develop progressive lens-induced myopia (LIM) and hyperopia (LIH). Retinal proteins were quantified with nano-liquid chromatography electrospray ionization coupled with tandem mass spectrometry (nanoLC-ESI-MS/MS). Bioinformatics analysis of differentially regulated proteins revealed that the majority originated from the cytoplasmic region and were related to various metabolic, glycolytic, or oxidative processes. The fold changes of four proteins of interest (vimentin, apolipoprotein A1, interphotoreceptor retinoid binding protein, and glutathione S-transferase) were further confirmed by a novel high-resolution multiple reaction monitoring mass spectrometry (MRM-HR) using a label-free approach. SIGNIFICANCE: Discovery of effective protein biomarkers of myopia has been extensively studied to inhibit myopia progression. This study first applied lens-induced hyperopia and myopia in the same chick to maximize the inter-ocular differences, aiming to discover more protein biomarker candidates. The findings provided new evidence that myopia was metabolism related, accompanied by altered energy generation and oxidative stress at retinal protein levels. The results in the retina were also compared to our previous study in vitreous using ICPL quantitative technology. We have now presented the protein changes in these two adjacent tissues, which may provide extra information of protein changes during ocular growth in myopia.
近视是视力受损最常见的原因，由于眼轴伸长，可能导致威胁视力的疾病或眼部并发症。近视发展的确切机制受到了广泛关注，对这些机制的理解对于临床预防或治疗非常必要。在这项研究中，我们应用同位素编码蛋白质标记（ICPL）的定量蛋白质组学技术，来鉴定近视鸡眼中差异调节的蛋白质，并从这些蛋白质的存在推断出过度眼轴伸长的可能发病机制。我们将 15 只刚孵化的白来航鸡（n = 15）的双眼分别戴上-10D 和 +10D 透镜，持续 3 天和 7 天，以分别诱导进展性透镜诱导性近视（LIM）和远视（LIH）。使用纳升液相色谱-电喷雾电离串联质谱（nanoLC-ESI-MS/MS）定量视网膜蛋白质。对差异调节蛋白质的生物信息学分析表明，大部分蛋白质来源于细胞质区域，与各种代谢、糖酵解或氧化过程有关。使用无标记方法的新型高分辨率多重反应监测质谱（MRM-HR）进一步证实了 4 种感兴趣蛋白质（波形蛋白、载脂蛋白 A1、光感受器间视网膜结合蛋白和谷胱甘肽 S-转移酶）的倍数变化。意义：广泛研究了发现有效的近视蛋白生物标志物，以抑制近视进展。本研究首次在同一只鸡中应用诱导性远视和近视，以最大限度地增加眼间差异，旨在发现更多的蛋白质生物标志物候选物。研究结果提供了新的证据，表明近视与代谢有关，视网膜蛋白质水平的能量生成和氧化应激发生改变。该结果还与我们之前使用 ICPL 定量技术在玻璃体中的研究进行了比较。我们现在已经展示了这两种相邻组织中的蛋白质变化，这可能为近视期间眼球生长过程中的蛋白质变化提供更多信息。
