Inhibition of heat shock protein 47 suppressed collagen production in Tenon’s capsule fibroblasts
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Abstract
Introduction. Glaucoma is the leading cause of irreversible blindness worldwide, and conjunctival bleb scarring remains the most frequent reason for the failure of glaucoma filtration surgery. Excessive proliferation of fibroblasts from Tenon’s capsule and excessive deposition of collagen contribute to the scarification of the conjunctival bleb. Heat shock protein 47 (HSP47) is assumed to act as a collagen-specific molecular chaperone, and thereby involved in the pathogenesis of fibrotic diseases. Therefore, we investigated the effect of HSP47 knockout against collagen type I (COLI) production in rat tenon’s fibroblasts.
Material and methods. Newborn rat tenon’s fibroblasts were cultured and verified by anti-vimentin antibody. Transfection efficiency of small interference RNA targeted against HSP47 was confirmed by quantitative real-time polymerase chain reaction (RT-qPCR) at 48 h after siRNA transfection and by western blot at 72 h after transfection. The mRNA and protein expression of HSP 47 and COLI were detected by RT-qPCR and western blot. The proliferation of cells was measured by cell counting kit-8 assay.
Results. HSP47 siRNA down-regulated the mRNA and protein levels of HSP47 in rat Tenon’s fibroblasts, and suppressed the mRNA and protein expression of COLI. Moreover, HSP47 siRNA had no significant effect on proliferation of rat Tenon’s fibroblasts.
Conclusions. HSP47 siRNA inhibits the production of COLI in rat Tenon’s fibroblasts, and may be the potential therapeutic method in bleb scarring after glaucoma filtration surgery.
Keywords: heat shock protein 47collagenTenon’s capsulefibroblastglaucomascar
References
- Vergroesen JE, Schuster AK, Stuart KV, et al. European Eye Epidemiology Consortium. Association of systemic medication use with glaucoma and intraocular pressure: the european eye epidemiology consortium. Ophthalmology. 2023; 130(9): 893–906.
- Kalarn S, Le T, Rhee DJ. The role of trabeculectomy in the era of minimally invasive glaucoma surgery. Curr Opin Ophthalmol. 2022; 33(2): 112–118.
- Igarashi N, Honjo M, Aihara M. Effects of mammalian target of rapamycin inhibitors on fibrosis after trabeculectomy. Exp Eye Res. 2021; 203: 108421.
- Wolters JEJ, van Mechelen RJS, Al Majidi R, et al. History, presence, and future of mitomycin C in glaucoma filtration surgery. Curr Opin Ophthalmol. 2021; 32(2): 148–159.
- Fluorouracil Filtering Surgery Study Group. Fluorouracil filtering surgery study one-year follow-up. Am J Ophthalmol. 2018; 186: xxxiii–xxxlii.
- Okuno D, Sakamoto N, Tagod MSO, et al. Screening of inhibitors targeting heat shock protein 47 involved in the development of idiopathic pulmonary fibrosis. ChemMedChem. 2021; 16(16): 2515–2523.
- Bellaye PS, Burgy O, Bonniaud P, et al. HSP47: a potential target for fibrotic diseases and implications for therapy. Expert Opin Ther Targets. 2021; 25(1): 49–62.
- Razzaque MS, Foster CS, Ahmed AR. Role of collagen-binding heat shock protein 47 and transforming growth factor-beta1 in conjunctival scarring in ocular cicatricial pemphigoid. Invest Ophthalmol Vis Sci. 2003; 44(4): 1616–1621.
- Hong S, Park K, Kim JH, et al. Role of heat shock protein 47 in transdifferentiation of human tenon's fibroblasts to myofibroblasts. BMC Ophthalmol. 2012; 12: 49.
- Guo Q, Tian Q, Tian X, et al. Effect of regulating the expression of HSP47 on collagen metabolism in scleral fibroblasts. Curr Eye Res. 2021; 46(3): 408–416.
- Wang W, Zhao J, Yang G, et al. Heat shock protein 47 is up-regulated in conjunctival bleb scarring after filtration surgery in rats. Graefes Arch Clin Exp Ophthalmol. 2015; 253(1): 91–97.
- Wang WW, Yu HZ, Yang X, et al. High levels of heat shock protein 47 in the aqueous humor of patients with acute primary angle closure. Ophthalmic Res. 2022 [Epub ahead of print].
- Xiong G, Chen J, Zhang G, et al. Hsp47 promotes cancer metastasis by enhancing collagen-dependent cancer cell-platelet interaction. Proc Natl Acad Sci U S A. 2020; 117(7): 3748–3758.
- Yoneda A, Minomi K, Tamura Y. HSP47 promotes metastasis of breast cancer by interacting with myosin IIA via the unfolded protein response transducer IRE1α. Oncogene. 2020; 39(23): 4519–4537.
- Lawitz EJ, Shevell DE, Tirucherai GS, et al. BMS-986263 in patients with advanced hepatic fibrosis: 36-week results from a randomized, placebo-controlled phase 2 trial. Hepatology. 2022; 75(4): 912–923.
- Wang F, Qi LX, Su Y, et al. Inhibition of cell proliferation of Tenon's capsule fibroblast by S-phase kinase-interacting protein 2 targeting SiRNA through increasing p27 protein level. Invest Ophthalmol Vis Sci. 2010; 51(3): 1475–1482.
- De Falco E, Scafetta G, Napoletano C, et al. A standardized laboratory and surgical method for in vitro culture isolation and expansion of primary human Tenon's fibroblasts. Cell Tissue Bank. 2013; 14(2): 277–287.
- Md Noh SM, Sheikh Abdul Kadir SH, Bannur ZM, et al. Effects of ranibizumab on the extracellular matrix production by human Tenon's fibroblast. Exp Eye Res. 2014; 127: 236–242.
- Addicks EM, Quigley HA, Green WR, et al. Histologic characteristics of filtering blebs in glaucomatous eyes. Arch Ophthalmol. 1983; 101(5): 795–798.
- Yamanaka O, Saika S, Ikeda K, et al. Connective tissue growth factor modulates extracellular matrix production in human subconjunctival fibroblasts and their proliferation and migration in vitro. Jpn J Ophthalmol. 2008; 52(1): 8–15.
- Zhang F, Liu Ke, Pan Z, et al. Effects of rosiglitazone/PHBV drug delivery system on postoperative fibrosis in rabbit glaucoma filtration surgery model. Drug Deliv. 2019; 26(1): 812–819.
- Liang L, Wang X, Zheng Y, et al. All‑trans‑retinoic acid modulates TGF‑β‑induced apoptosis, proliferation, migration and extracellular matrix synthesis of conjunctival fibroblasts by inhibiting PI3K/AKT signaling. Mol Med Rep. 2019; 20(3): 2929–2935.
- Sato Y, Murase K, Kato J, et al. Resolution of liver cirrhosis using vitamin A-coupled liposomes to deliver siRNA against a collagen-specific chaperone. Nat Biotechnol. 2008; 26(4): 431–442.
- Yamakawa T, Ohigashi H, Hashimoto D, et al. Vitamin A-coupled liposomes containing siRNA against HSP47 ameliorate skin fibrosis in chronic graft-versus-host disease. Blood. 2018; 131(13): 1476–1485.
- Ohigashi H, Hashimoto D, Hayase E, et al. Ocular instillation of vitamin A-coupled liposomes containing HSP47 siRNA ameliorates dry eye syndrome in chronic GVHD. Blood Adv. 2019; 3(7): 1003–1010.
- Liu Y, Liu J, Quimbo A, et al. Anti-HSP47 siRNA lipid nanoparticle ND-L02-s0201 reverses interstitial pulmonary fibrosis in preclinical rat models. ERJ Open Res. 2021; 7(2).
- Tang X, Liu L, Liu S, et al. MicroRNA-29a inhibits collagen expression and induces apoptosis in human fetal scleral fibroblasts by targeting the Hsp47/Smad3 signaling pathway. Exp Eye Res. 2022; 225: 109275.
