Age-Related Dysregulation of α5β1 and αvβ3 Integrin Activity Alters Contractile Properties of Trabecular Meshwork Cells.

IF 5 2区 医学 Q1 OPHTHALMOLOGY
Kassidy L Johns, Jennifer A Faralli, Mark S Filla, Nandini S Shah, Ying Ying Sun, Kate E Keller, Donna M Peters
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Abstract

Purpose: Age and elevated intraocular pressure are major risk factors for primary open-angle glaucoma (POAG) which is caused by a restriction in aqueous humor outflow from the anterior chamber. In this study, we investigated whether age-related changes in integrin subunit expression and activity signifies an early event in initiating fibrotic-like changes in the TM that could restrict outflow.

Methods: Human trabecular meshwork (TM) cells from young (<40 years) and old (>50 years) donor eyes were used. Flow cytometry, RT-qPCR, and immunofluorescence microscopy were used to evaluate levels of integrin and αSMA expression. On-cell westerns were used to determine fibronectin levels. Collagen gel contraction assays were used to determine contractile properties of cells and shRNA was used to knockdown α5β1 integrin levels.

Results: Studies revealed a significant decrease in α5 integrin expression in TM cells from older individuals. This loss was accompanied by an increase in activated but not total αvβ3 integrin levels. TM cells from older donors expressed higher levels of αSMA mRNA, assembled αSMA-containing stress fibers, and contracted collagen gels significantly more than young TM cells. TM cells from old donors also assembled higher levels of insoluble fibronectin fibrils and contained higher levels of EDB+ fibronectin in their extracellular matrix. shRNA knockdown of α5 integrin subunits showed that the increase in αvβ3 integrin activity was due to lower levels of α5 integrin expression.

Conclusions: These studies suggest that age-related dysregulation of α5β1 and αvβ3 integrin signaling may represent an important early molecular event in inducing fibrogenic pathways associated with POAG.

α5β1和αvβ3整合素活性的年龄相关失调改变小梁网细胞的收缩特性。
目的:年龄和眼压升高是原发性开角型青光眼(POAG)的主要危险因素,POAG是由房水流出受限引起的。在这项研究中,我们研究了整合素亚基表达和活性的年龄相关变化是否意味着在TM中启动纤维化样变化的早期事件,从而限制流出。方法:采用年轻(50岁)供眼的人小梁网(TM)细胞。流式细胞术、RT-qPCR和免疫荧光显微镜检测整合素和α - sma的表达水平。细胞内western检测纤维连接蛋白水平。用胶原凝胶收缩法测定细胞的收缩特性,用shRNA敲低α5β1整合素水平。结果:研究显示老年人TM细胞α5整合素表达明显降低。这种损失伴随着激活的αvβ3整合素水平的增加,而不是总αvβ3整合素水平的增加。老年供体TM细胞α - sma mRNA表达水平、α - sma应激纤维组装水平和胶原蛋白凝胶收缩水平均明显高于年轻供体TM细胞。来自老年供体的TM细胞也聚集了更高水平的不溶性纤维连接蛋白原纤维,并且在其细胞外基质中含有更高水平的EDB+纤维连接蛋白。shRNA敲低α5整合素亚基表明αvβ3整合素活性升高是由于α5整合素表达水平降低所致。结论:这些研究表明,α5β1和αvβ3整合素信号的年龄相关失调可能是诱导POAG相关纤维化途径的重要早期分子事件。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
6.90
自引率
4.50%
发文量
339
审稿时长
1 months
期刊介绍: Investigative Ophthalmology & Visual Science (IOVS), published as ready online, is a peer-reviewed academic journal of the Association for Research in Vision and Ophthalmology (ARVO). IOVS features original research, mostly pertaining to clinical and laboratory ophthalmology and vision research in general.
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