Role of Endogenous Galectin-3 on Cell Biology of Immortalized Retinal Pigment Epithelial Cells In Vitro

Liesenhoff, Caspar; Hillenmayer, Marlene; Havertz, Caroline; Geerlof, Arie; Hartmann, Daniela; Priglinger, Siegfried G.; Priglinger, Claudia S.; Ohlmann, Andreas (2025): Role of Endogenous Galectin-3 on Cell Biology of Immortalized Retinal Pigment Epithelial Cells In Vitro. International Journal of Molecular Sciences, 26 (15): 7622. ISSN 1422-0067

Abstract

Galectin-3 is a multifunctional protein that is associated with diseases of the chorioretinal interface, in which the retinal pigment epithelium (RPE) plays a central role in disease de-velopment and progression. Since galectin-3 can function extracellularly as well as intracel-lularly via different mechanisms, we developed an immortalized human RPE cell line (ARPE-19) with a knockdown for galectin-3 expression (ARPE-19/LGALS3+/−) using a sgRNA/Cas9 all-in-one expression vector. By Western blot analysis, a reduced galectin-3 expression of approximately 48 to 60% in heterozygous ARPE-19/LGALS3+/− cells was ob-served when compared to native controls. Furthermore, ARPE-19/LGALS3+/− cells dis-played a flattened, elongated phenotype with decreased E-cadherin as well as enhanced N-cadherin and α-smooth muscle actin mRNA expression, indicating an epithelial–mesenchy-mal transition of the cells. Compared to wildtype controls, ARPE-19/LGALS3+/− cells had significantly reduced metabolic activity to 86% and a substantially decreased proliferation to 73%. Furthermore, an enhanced cell adhesion and a diminished migration of immortal-ized galectin-3 knockdown RPE cells was observed compared to native ARPE-19 cells. Fi-nally, by Western blot analysis, reduced pAKT, pERK1/2, and β-catenin signaling were de-tected in ARPE-19/LGALS3+/− cells when compared to wildtype controls. In summary, in RPE cells, endogenous galectin-3 appears to be essential for maintaining the epithelial phe-notype as well as cell biological functions such as metabolism, proliferation, or migration, effects that might be mediated via a decreased activity of the AKT, ERK1/2, and β-catenin signaling pathways.

• Dokumententyp: Artikel (Klinikum der LMU)
• Organisationseinheit (Fakultäten): 07 Medizin > Klinikum der LMU München > Augenklinik und Poliklinik
• DFG-Fachsystematik der Wissenschaftsbereiche: Lebenswissenschaften
• DFG: Gefördert durch die Deutsche Forschungsgemeinschaft (DFG) - 181475910