MiR-18a-5p Targets Connective Tissue Growth Factor Expression and Inhibits Transforming Growth Factor β2-Induced Trabecular Meshwork Cell Contractility

Overview

Journal Genes (Basel)

Publisher MDPI

Date 2022 Aug 26

PMID 36011411

Authors

John Knox

George Bou-Gharios

Kevin J Hamill

Colin E Willoughby

Affiliations

Soon will be listed here.

Abstract

Increased trabecular meshwork (TM) cell and tissue contractility is a driver of the reduced outflow facility and elevation of intraocular pressure (IOP) associated with primary open-angle glaucoma (POAG). Connective tissue growth factor (CTGF) is an established mediator of TM cell contractility, and its expression is increased in POAG due to transforming growth factor β 2 (TGFβ2) signalling. Inhibiting CTGF upregulation using microRNA (miRNA) mimetics could represent a new treatment option for POAG. A combination of in silico predictive tools and a literature review identified a panel of putative CTGF-targeting miRNAs. Treatment of primary human TM cells with 5 ng/mL TGFβ2 for 24 h identified miR-18a-5p as a consistent responder, being upregulated in cells from five different human donors. Transfection of primary donor TM cells with 20 nM synthetic miR-18a-5p mimic reduced TGFβ2-induced CTGF protein expression, and stable lentiviral-mediated overexpression of this miRNA reduced TGFβ2-induced contraction of collagen gels. Together, these findings identify miR-18a-5p as a mediator of the TGFβ2 response and a candidate therapeutic agent for glaucoma via its ability to inhibit CTGF-associated increased TM contractility.

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References

Drewry M, Helwa I, Allingham R, Hauser M, Liu Y . miRNA Profile in Three Different Normal Human Ocular Tissues by miRNA-Seq. Invest Ophthalmol Vis Sci. 2016; 57(8):3731-9. PMC: 4961003. DOI: 10.1167/iovs.16-19155. View

Chou C, Shrestha S, Yang C, Chang N, Lin Y, Liao K . miRTarBase update 2018: a resource for experimentally validated microRNA-target interactions. Nucleic Acids Res. 2017; 46(D1):D296-D302. PMC: 5753222. DOI: 10.1093/nar/gkx1067. View

Keller K, Aga M, Bradley J, Kelley M, Acott T . Extracellular matrix turnover and outflow resistance. Exp Eye Res. 2008; 88(4):676-82. PMC: 2700052. DOI: 10.1016/j.exer.2008.11.023. View

Wormald R, Virgili G, Azuara-Blanco A . Systematic reviews and randomised controlled trials on open angle glaucoma. Eye (Lond). 2019; 34(1):161-167. PMC: 7002425. DOI: 10.1038/s41433-019-0687-5. View

Huntzinger E, Izaurralde E . Gene silencing by microRNAs: contributions of translational repression and mRNA decay. Nat Rev Genet. 2011; 12(2):99-110. DOI: 10.1038/nrg2936. View

Guo Y, Lu X, Wang H . Downregulation of miR-18a induces CTGF and promotes proliferation and migration of sodium hyaluronate treated human corneal epithelial cells. Gene. 2016; 591(1):129-136. DOI: 10.1016/j.gene.2016.07.008. View

Keller K, Bhattacharya S, Borras T, Brunner T, Chansangpetch S, Clark A . Consensus recommendations for trabecular meshwork cell isolation, characterization and culture. Exp Eye Res. 2018; 171:164-173. PMC: 6042513. DOI: 10.1016/j.exer.2018.03.001. View

Lee E, Han J, Park D, Cho J, Kee C . Effect of connective tissue growth factor gene editing using adeno-associated virus-mediated CRISPR-Cas9 on rabbit glaucoma filtering surgery outcomes. Gene Ther. 2020; 28(5):277-286. DOI: 10.1038/s41434-020-0166-4. View

Kern F, Krammes L, Danz K, Diener C, Kehl T, Kuchler O . Validation of human microRNA target pathways enables evaluation of target prediction tools. Nucleic Acids Res. 2020; 49(1):127-144. PMC: 7797041. DOI: 10.1093/nar/gkaa1161. View

10.

Sun D, Han S, Liu C, Zhou R, Sun W, Zhang Z . Microrna-199a-5p Functions as a Tumor Suppressor via Suppressing Connective Tissue Growth Factor (CTGF) in Follicular Thyroid Carcinoma. Med Sci Monit. 2016; 22:1210-7. PMC: 4830201. DOI: 10.12659/msm.895788. View

11.

van Almen G, Verhesen W, van Leeuwen R, van de Vrie M, Eurlings C, Schellings M . MicroRNA-18 and microRNA-19 regulate CTGF and TSP-1 expression in age-related heart failure. Aging Cell. 2011; 10(5):769-79. PMC: 3193380. DOI: 10.1111/j.1474-9726.2011.00714.x. View

12.

Tomarev S, Wistow G, Raymond V, Dubois S, Malyukova I . Gene expression profile of the human trabecular meshwork: NEIBank sequence tag analysis. Invest Ophthalmol Vis Sci. 2003; 44(6):2588-96. DOI: 10.1167/iovs.02-1099. View

13.

Li G, Luna C, Qiu J, Epstein D, Gonzalez P . Alterations in microRNA expression in stress-induced cellular senescence. Mech Ageing Dev. 2009; 130(11-12):731-41. PMC: 2795064. DOI: 10.1016/j.mad.2009.09.002. View

14.

Kehl T, Backes C, Kern F, Fehlmann T, Ludwig N, Meese E . About miRNAs, miRNA seeds, target genes and target pathways. Oncotarget. 2018; 8(63):107167-107175. PMC: 5739805. DOI: 10.18632/oncotarget.22363. View

15.

John B, Enright A, Aravin A, Tuschl T, Sander C, Marks D . Human MicroRNA targets. PLoS Biol. 2004; 2(11):e363. PMC: 521178. DOI: 10.1371/journal.pbio.0020363. View

16.

Greene K, Stamer W, Liu Y . The role of microRNAs in glaucoma. Exp Eye Res. 2021; 215:108909. PMC: 8923961. DOI: 10.1016/j.exer.2021.108909. View

17.

Wiederholt M . Direct involvement of trabecular meshwork in the regulation of aqueous humor outflow. Curr Opin Ophthalmol. 1998; 9(2):46-9. DOI: 10.1097/00055735-199804000-00009. View

18.

Zhang Q, Ye H, Xiang F, Song L, Zhou L, Cai P . miR-18a-5p Inhibits Sub-pleural Pulmonary Fibrosis by Targeting TGF-β Receptor II. Mol Ther. 2017; 25(3):728-738. PMC: 5363213. DOI: 10.1016/j.ymthe.2016.12.017. View

19.

Lun W, Wu X, Deng Q, Zhi F . MiR-218 regulates epithelial-mesenchymal transition and angiogenesis in colorectal cancer via targeting CTGF. Cancer Cell Int. 2018; 18:83. PMC: 5994014. DOI: 10.1186/s12935-018-0575-2. View

20.

Stahl F, Wiederholt M . Differential smooth muscle-like contractile properties of trabecular meshwork and ciliary muscle. Exp Eye Res. 1991; 53(1):33-8. DOI: 10.1016/0014-4835(91)90141-z. View