MicroRNA-29b targets ADAM12 and 19 to regulate the extracellular matrix in lamina cribrosa cells.

IF 2.9 3区 生物学 Q3 CELL BIOLOGY
Aoife Smyth, Breedge Callaghan, Mustapha Irnaten, Darrell Andrews, Colin E Willoughby, Colm O'Brien
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Abstract

Glaucoma remains the leading cause of irreversible blindness worldwide, with elevated intraocular pressure (IOP) being the only modifiable risk factor in primary open-angle glaucoma (POAG). Despite adequate IOP control, many patients continue to progress to irreversible optic neuropathy, emphasising the need for alternate treatments. Transforming growth factor-beta (TGF-β) promotes extracellular matrix (ECM) production and fibrosis at the optic nerve head (ONH) in glaucoma. A disintegrin and metalloprotease-12 and metalloprotease-19 (ADAM12 and ADAM19) are implicated in fibrosis. Recent studies have explored miRNA-based manipulation of the TGF-β signalling pathway as a potential therapeutic strategy in fibrosis. This study investigates whether miR-29b modulation affects ADAM12, ADAM19, and ECM gene expression in human lamina cribrosa (LC) cells. Primary human normal lamina cribrosa (NLC) and glaucoma LC (GLC) cells were treated with TGF-β1 and transfected with either a miR-29b mimic or control. Gene expression levels of ADAM12, ADAM19, miR-29b, and several ECM genes were quantified using real-time RT-qPCR, and protein expression levels by Western blotting. ADAM12 and ADAM19 expression was elevated in untreated GLC cells, and treatment with TGF-β1 in both NLC and GLC cells increased ADAM12 and ADAM19 expression. The expression of miR-29b was significantly reduced in both GLC- and TGF-β1-treated NLC and GLC cells. Transfection with miR-29b resulted in a marked reduction in ADAM12 and ADAM19 mRNA expression in TGF-β1-treated NLC and GLC cells. Additionally, miR-29b transfection reduced ECM gene expression in both NLC and GLC under TGF-β1 stimulation. Our results demonstrate that miR-29b plays a crucial role in fibrotic remodelling at the LC by antagonising the effects of TGF-β1 on ADAM and ECM gene expression, representing a novel therapeutic target in glaucoma.

MicroRNA-29b靶向ADAM12和adam19调控筛板细胞的细胞外基质。
青光眼仍然是世界范围内不可逆失明的主要原因,眼压升高是原发性开角型青光眼(POAG)唯一可改变的危险因素。尽管充分的IOP控制,许多患者继续发展为不可逆的视神经病变,强调需要替代治疗。转化生长因子-β (TGF-β)促进青光眼视神经头(ONH)细胞外基质(ECM)的产生和纤维化。崩解素和金属蛋白酶-12和金属蛋白酶-19 (ADAM12和ADAM19)与纤维化有关。最近的研究已经探索了基于mirna的TGF-β信号通路的操纵作为纤维化的潜在治疗策略。本研究探讨miR-29b调节是否影响人筛板(LC)细胞ADAM12、ADAM19和ECM基因的表达。用TGF-β1处理原代人正常滤膜(NLC)和青光眼LC (GLC)细胞,转染miR-29b模拟物或对照物。实时RT-qPCR检测ADAM12、ADAM19、miR-29b和多个ECM基因的基因表达水平,Western blotting检测蛋白表达水平。未处理GLC细胞ADAM12和ADAM19表达升高,TGF-β1处理NLC和GLC细胞ADAM12和ADAM19表达均升高。在GLC-和TGF-β1处理的NLC和GLC细胞中,miR-29b的表达均显著降低。转染miR-29b可显著降低TGF-β1处理的NLC和GLC细胞中ADAM12和ADAM19 mRNA的表达。此外,在TGF-β1刺激下,转染miR-29b可降低NLC和GLC中ECM基因的表达。我们的研究结果表明,miR-29b通过拮抗TGF-β1对ADAM和ECM基因表达的影响,在LC的纤维化重塑中起着至关重要的作用,代表了青光眼的新治疗靶点。
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
Cell and Tissue Research
Cell and Tissue Research 生物-细胞生物学
CiteScore
7.00
自引率
2.80%
发文量
142
审稿时长
1 months
期刊介绍: The journal publishes regular articles and reviews in the areas of molecular, cell, and supracellular biology. In particular, the journal intends to provide a forum for publishing data that analyze the supracellular, integrative actions of gene products and their impact on the formation of tissue structure and function. Submission of papers with an emphasis on structure-function relationships as revealed by recombinant molecular technologies is especially encouraged. Areas of research with a long-standing tradition of publishing in Cell & Tissue Research include: - neurobiology - neuroendocrinology - endocrinology - reproductive biology - skeletal and immune systems - development - stem cells - muscle biology.
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