JM-USDA Human Nutrition Research Center on Aging, Tufts University, Boston, MA, USA; School of Health Sciences and Veterinary School, Universidad Cardenal Herrera-CEU, CEU Universities, Moncada, Valencia, Spain.
JM-USDA Human Nutrition Research Center on Aging, Tufts University, Boston, MA, USA.
Redox Biol. 2023 Oct;66:102869. doi: 10.1016/j.redox.2023.102869. Epub 2023 Sep 1.
The lens proteome undergoes dramatic composition changes during development and maturation. A defective developmental process leads to congenital cataracts that account for about 30% of cases of childhood blindness. Gene mutations are associated with approximately 50% of early-onset forms of lens opacity, with the remainder being of unknown etiology. To gain a better understanding of cataractogenesis, we utilized a transgenic mouse model expressing a mutant ubiquitin protein in the lens (K6W-Ub) that recapitulates most of the early pathological changes seen in human congenital cataracts. We performed mass spectrometry-based tandem-mass-tag quantitative proteomics in E15, P1, and P30 control or K6W-Ub lenses. Our analysis identified targets that are required for early normal differentiation steps and altered in cataractous lenses, particularly metabolic pathways involving glutathione and amino acids. Computational molecular phenotyping revealed that glutathione and taurine were spatially altered in the K6W-Ub cataractous lens. High-performance liquid chromatography revealed that both taurine and the ratio of reduced glutathione to oxidized glutathione, two indicators of redox status, were differentially compromised in lens biology. In sum, our research documents that dynamic proteome changes in a mouse model of congenital cataracts impact redox biology in lens. Our findings shed light on the molecular mechanisms associated with congenital cataracts and point out that unbalanced redox status due to reduced levels of taurine and glutathione, metabolites already linked to age-related cataract, could be a major underlying mechanism behind lens opacities that appear early in life.
晶状体蛋白质组在发育和成熟过程中经历了显著的组成变化。发育过程的缺陷会导致先天性白内障,约占儿童失明病例的 30%。基因突变与大约 50%的早发性晶状体混浊有关,其余的病因不明。为了更好地了解白内障的发生机制,我们利用在晶状体中表达突变泛素蛋白的转基因小鼠模型(K6W-Ub),该模型重现了人类先天性白内障中所见的大多数早期病理变化。我们对 E15、P1 和 P30 对照或 K6W-Ub 晶状体进行了基于质谱的串联质量标签定量蛋白质组学分析。我们的分析确定了需要早期正常分化步骤的靶标,以及在白内障晶状体中改变的靶标,特别是涉及谷胱甘肽和氨基酸的代谢途径。计算分子表型分析显示,谷胱甘肽和牛磺酸在 K6W-Ub 白内障晶状体中空间改变。高效液相色谱法显示,牛磺酸和还原型谷胱甘肽与氧化型谷胱甘肽的比值(两种氧化还原状态的指标)在晶状体生物学中均有差异受损。总之,我们的研究表明,先天性白内障小鼠模型中的动态蛋白质组变化会影响晶状体的氧化还原生物学。我们的研究结果揭示了与先天性白内障相关的分子机制,并指出由于牛磺酸和谷胱甘肽水平降低导致的不平衡氧化还原状态,这两种代谢物已经与年龄相关性白内障有关,可能是生命早期出现晶状体混浊的主要潜在机制。