Complement-mediated release of fibroblast growth factor 2 from human RPE cells. (March 2021)

• Record Type: Journal Article
• Authors: Yang, Ping; Neal, Samantha E.; Buehne, Kristen L.; Tewkesbury, Grace M.; Klingeborn, Mikael; Yang, Yae-Young; Baciu, Peter; Jaffe, Glenn J.
• Is Part Of: Experimental eye research. Volume 204(2021)
• Publication Date: 2021-03
• Subjects: Age-related macular degeneration -- Retinal pigment epithelial -- Complement -- Fibroblast growth factor 2; Ophthalmology -- Periodicals; Eye -- Periodicals; Œil -- Périodiques; Ophthalmology; Periodicals; Electronic journals; 612.8405
• DOI: 10.1016/j.exer.2021.108471
• Languages: English
• ISSNs: 0014-4835

Abstract

Abstract: Purpose: Complement activation is associated with choroidal neovascularization (CNV) in age-related macular degeneration (AMD). Fibroblast growth factor 2 (FGF2) and membrane attack complex (MAC) are present in eyes of patients with CNV. Herein, we investigated the effect of complement activation on FGF2 release in human retinal pigment epithelial (RPE) cells. Methods: Cultured human RPE cells were primed with an anti-RPE antibody and then treated with C1q-depleted human serum in the presence or absence of Tec kinases inhibitor (LFM-A13). 38 cytokines/chemokines levels were measured by Luminex technology. Secretion of FGF2 and interleukin (IL)-6 was assessed by ELISA. Tec protein was measured by Western blot. mRNA expression of FGF2, chemokine (C-X-C motif) ligand 1 (CXCL-1), and family members of Tec kinases was evaluated by qPCR. Cell viability and MAC deposition were determined by WST-1 assay and flow cytometry, respectively. Results: Complement activation caused increased FGF2 and IL-6 release. FGF2 was released when C6-depleted human serum was reconstituted with C6. Anti-C5 antibody significantly attenuated complement-mediated FGF2 release, but not IL-6. FGF2 mRNA levels were not affected, while CXCL-1 mRNA levels were increased by complement activation. FGF2-containing extracellular vesicles were detected in response to complement challenge. Tec mRNA and protein were expressed in RPE cells. In the presence of LFM-A13, secretion of FGF2, but not IL-6, and MAC deposition were significantly decreased and cell viability was significantly increased in complement-treated cells when compared to controls. Conclusions: Complement plays an important role to release FGF2 from RPE cells. Tec kinase is involved in MAC formation and complement-mediated FGF2 release. This information suggests a role for complement activation to mediate neovascularization in conditions such as AMD, and may elucidate potential therapeutic targets. Highlights: Complement activation increases RPE cell FGF2 release FGF2 release is dependent on MAC formation, but not endogenous synthesis Complement activation releases FGF2-containing extracellular vesicles RPE cells express Tec kinase Inhibition of Tec kinase reduces complement-mediated FGF2 release and MAC formation