Establishment of human corneal epithelial organoids for ex vivo modelling dry eye disease

Abstract

Dry eye disease (DED) is a growing public health concern affecting millions of people worldwide and causing ocular discomfort and visual disturbance. Developing its therapeutic drugs based on animal models suffer from interspecies differences and poor prediction of human trials. Here, we established long-term 3D human corneal epithelial organoids, which recapitulated the cell lineages and gene expression signature of the human corneal epithelium. Organoids can be regulated to differentiate ex vivo, but the addition of FGF10 inhibits this process. In the hyperosmolar-induced DED organoid model, the release of inflammatory factors increased, resulting in damage to the stemness of stem cells and a decrease in functional mucin 1 protein. Furthermore, we found that the organoids could mimic clinical drug treatment responses, suggesting that corneal epithelial organoids are promising candidates for establishing a drug testing platform ex vivo. In summary, we established a functional, long-term 3D human epithelial organoid that may serve as an ex vivo model for studying the functional regulation and disease modelling.

References

[1] AbdalkaderRK, FujitaT. Corneal epithelium models for safety assessment in drug development: present and future directions. Exp Eye Res. 2023;237:109697.

[2] YazdanpanahG, Jabbehdari S, DjalilianAR. Limbal and corneal epithelial homeostasis. Curr Opin Ophthalmol. 2017;28(4):348-354.

[3] StapletonF, AlvesM, BunyaVY, et al. TFOS DEWS II epidemiology report. Ocul Surf. 2017;15(3):334-365.

[4] ClaytonJA. Dry eye. N Engl J Med. 2018;378(23):2212-2223.

[5] BedosL, Wickham H, GabrielV, et al. Culture and characterization of canine and feline corneal epithelial organoids: a new tool for the study and treatment of corneal diseases. Front Vet Sci. 2022;9:1050467.

[6] RubelowskiAK, LattaL, KatiyarP, et al. HCE-T cell line lacks cornea-specific differentiation markers compared to primary limbal epithelial cells and differentiated corneal epithelium. Graefes Arch Clin Exp Ophthalmol. 2020;258(3):565-575.

[7] GarciaI, Capallere C, ArcioniM, et al. Establishment and performance assessment of an in-house 3D reconstructed human cornea-like epithelium (RhCE) as a screening tool for the identification of liquid chemicals with potential eye hazard. Toxicol In Vitro. 2019;61:104604.

[8] CleversH. Modeling development and disease with organoids. Cell. 2016;165(7):1586-1597.

[9] Bannier-HélaouëtM, KorvingJ, MaZ, et al. Human conjunctiva organoids to study ocular surface homeostasis and disease. Cell Stem Cell. 2024;31(2):227-243.e12.

[10] SwarupA, Phansalkar R, MorriM, et al. Single-cell transcriptomic analysis of corneal organoids during development. Stem Cell Rep. 2023;18(12):2482-2497.

[11] FosterJW, WahlinK, AdamsSM, Birk DE, ZackDJ, ChakravartiS. Cornea organoids from human induced pluripotent stem cells. Sci Rep. 2017;7:41286.

[12] HigaK, Higuchi J, KimotoR, et al. Human corneal limbal organoids maintaining limbal stem cell niche function. Stem Cell Res. 2020;49:102012.

[13] Bannier-HélaouëtM, PostY, Korving J, et al. Exploring the human lacrimal gland using organoids and single-cell sequencing. Cell Stem Cell. 2021;28(7):1221-1232.e7.

[14] GruebM, Bartz-Schmidt KU, RohrbachJM. Adrenergic regulation of cAMP/protein kinase a pathway in corneal epithelium and endothelium. Ophthalmic Res. 2008;40(6):322-328.

[15] TiwariA, Swamynathan S, CampbellG, JhanjiV, Swamynathan SK. BMP6 regulates corneal epithelial cell stratification by coordinating their proliferation and differentiation and is upregulated in pterygium. Invest Ophthalmol Vis Sci. 2020;61(10):46.

[16] LattaL, Viestenz A, StachonT, et al. Human aniridia limbal epithelial cells lack expression of keratins K3 and K12. Exp Eye Res. 2018;167:100-109.

[17] GuoP, SunH, ZhangY, et al. Limbal niche cells are a potent resource of adult mesenchymal progenitors. J Cell Mol Med. 2018;22(7):3315-3322.

[18] BronAJ, de Paiva CS, ChauhanSK, et al. TFOS DEWS II pathophysiology report. Ocul Surf. 2017;15(3):438-510.

[19] WilsonSL, Ahearne M, HopkinsonA. An overview of current techniques for ocular toxicity testing. Toxicology. 2015;327:32-46.

[20] Kado AbdalkaderR, Chaleckis R, FujitaT, KameiKI. Modeling dry eye with an air–liquid interface in corneal epithelium-on-a-chip. Sci Rep. 2024;14(1):4185.

[21] MessmerEM, von Lindenfels V, GarbeA, KampikA. Matrix metalloproteinase 9 testing in dry eye disease using a commercially available point-of-care immunoassay. Ophthalmology. 2016;123(11):2300-2308.

[22] KeatingGM. Diquafosol ophthalmic solution 3%: a review of its use in dry eye. Drugs. 2015;75(8):911-922.

[23] ParkJH, MoonSH, KangDH, et al. Diquafosol sodium inhibits apoptosis and inflammation of corneal epithelial cells via activation of Erk1/2 and RSK: in vitro and in vivo dry eye model. Invest Ophthalmol Vis Sci. 2018;59(12):5108-5115.

[24] JonesL, DownieLE, KorbD, et al. TFOS DEWS II management and therapy report. Ocul Surf. 2017;15(3):575-628.

[25] ZhaoX, JiaQ, FuJ. Effectiveness of rb-bFGF eye drops for post-cataract surgery dry eye and observation of changes in tear secretion and corneal damage in patients. Altern Ther Health Med. 2023;29(8):489-495.

[26] EomY, SongJS, KimHM. Effectiveness of topical Cyclosporin A 0.1%, diquafosol tetrasodium 3%, and their combination, in dry eye disease. J Ocul Pharmacol Ther. 2022;38(10):682-694.

[27] KratochvilMJ, Seymour AJ, LiTL, PaşcaSP, KuoCJ, HeilshornSC. Engineered materials for organoid systems. Nat Rev Mater. 2019;4(9):606-622.

[28] KozlowskiMT, CrookCJ, KuHT. Towards organoid culture without Matrigel. Commun Biol. 2021;4(1):1387.

[29] FinburghEN, Mauduit O, NoguchiT, et al. Role of FGF10/FGFR2b signaling in homeostasis and regeneration of adult lacrimal gland and corneal epithelium proliferation. Invest Ophthalmol Vis Sci. 2023;64(1):21.

[30] RahmanMM, KimDH, ParkCK, Kim YH. Experimental models, induction protocols, and measured parameters in dry eye disease: focusing on practical implications for experimental research. Int J Mol Sci. 2021;22(22):12102.

[31] ParkB, JoK, LeeTG, Hyun SW, KimJS, KimCS. Polydatin inhibits NLRP3 inflammasome in dry eye disease by attenuating oxidative stress and inhibiting the NF-κB pathway. Nutrients. 2019;11(11):2792.

[32] LiuZ, ChenD, ChenX, et al. Autophagy activation protects ocular surface from inflammation in a dry eye model in vitro. Int J Mol Sci. 2020;21(23):8966.

[33] MeloniM, Carriero F, CeriottiL, BarabinoS. Development of a novel in vitro Immuno-competent model of dry eye disease and its use to evaluate the efficacy of an ocular surface modulator. Ocul Immunol Inflamm. 2022;30(7–8):1816-1824.

[34] VoßS, Behrmann T, ReichlS. Development of in vitro dry eye models to study proliferative and anti-inflammatory effects of allogeneic serum eye drops. Int J Mol Sci. 2023;24(2):1567.

[35] RenY, LuH, ReinachPS, et al. Hyperosmolarity-induced AQP5 upregulation promotes inflammation and cell death via JNK1/2 activation in human corneal epithelial cells. Sci Rep. 2017;7(1):4727.

[36] YamaguchiT. Inflammatory response in dry eye. Invest Ophthalmol Vis Sci. 2018;59(14):des192-des199.

[37] LinS, CaiM, ZhangL, et al. Limbal stem cell dysfunction induced by severe dry eye via activation of the p38 MAPK signaling pathway. Am J Pathol. 2023;193(11):1863-1878.

[38] RushSW, ChainJ, DasH. Corneal epithelial stem cell supernatant in the treatment of severe dry eye disease: a pilot study. Clin Ophthalmol. 2021;15:3097-3107.

[39] de PaivaCS, Pflugfelder SC, NgSM, AkpekEK. Topical cyclosporine a therapy for dry eye syndrome. Cochrane Database Syst Rev. 2019;9(9):Cd010051.

[40] WatanabeN, Santostefano KE, YachnisAT, TeradaN. A pathologist’s perspective on induced pluripotent stem cells. Lab Invest. 2017;97(10):1126-1132.

RIGHTS & PERMISSIONS

2024 The Author(s). Cell Proliferation published by Beijing Institute for Stem Cell and Regenerative Medicine and John Wiley & Sons Ltd.