Galectin-3 Enhances Migration of Minature Pig Bone Marrow Mesenchymal Stem Cells Through Inhibition of RhoA-GTP Activity

Abstract

Bone marrow mesenchymal stem cells (BM-MSCs) are used in tissue engineering because of their migration characters. However, BM-MSCs have limitations in terms of reaching injuries and self-renewal. Therefore, enhancement of BM-MSC migration is important for therapeutic applications. Here, we assessed whether galectin-3 (Gal-3) increases the migration of minature pig BM-MSCs. Gal-3 was knocked down by short hairpin RNA (shRNA) or overexpressed using a lentiviral vector in Wuzhishan minature pig BM-MSCs. Proliferation and migration assays showed that knockdown of Gal-3 impaired BM-MSC proliferation and migration, whereas Gal-3 overexpression promoted these behaviors. RhoA-GTP activity was upregulated in Gal-3 shRNA-transfected BM-MSCs, while Rac-1- and Cdc42-GTP showed no changes. Western blotting indicated downregulation of p-AKT (ser473) and p-Erk1/2 after serum starvation for 12 h in Gal-3-knockdown BM-MSCs. p-AKT (ser473) expression was upregulated after serum starvation for 6 h, and p-Erk1/2 expression was unchanged in Gal-3-overexpressing BM-MSCs. Treatment with C3 transferase or Y27632 enhanced migration, whereas Gal-3 knockdown impaired migration in treated cells. These results demonstrate that Gal-3 may enhance BM-MSC migration, mainly through inhibiting RhoA-GTP activity, increasing p-AKT (ser473) expression, and regulating p-Erk1/2 levels. Our study suggests a novel function of Gal-3 in regulating minature pig BM-MSC migration, which may be beneficial for therapeutic applications.

Galectin-3 Knockdown Impairs Survival, Migration, and Immunomodulatory Actions of Mesenchymal Stromal Cells in a Mouse Model of Chagas Disease Cardiomyopathy

“…Gal-3 was recently found to promote migration of MSC through inhibition of RhoA-GTP activity, enhancement of p-AKT (ser473) expression, and regulation of p-Erk1/2 levels [29]. Based on these data and in our findings, it is reasonable to suggest that Gal-3 plays a significant role in MSC migration and homing, which could have several implications in a cell transplantation setting.…”

Role of MSC-derived galectin 3 in the AML microenvironment

“…A role for LGALS3 as a positive regulator of AKT is well established (36, 37), and LGALS3's regulation of AKT appears to be important for migration in pig MSC (38). We examined AKT expression and AKT phosphorylation in MSC transduced with control lentivirus and MSC transduced with LGALS3 shRNA and found phosphorylation of AKT at serine 473 was reduced by roughly half (Figure 4A).…”

“…Suppression of LGALS3 in MSC reduces MSC abilit y to inhibit T cell function (24, 44). Recent studies have suggested LGALS3 plays an important role in MSC migration (38, 47). A study of LGALS3 in murine MSC revealed that the galectin is elevated in MSC as mice age (48).…”

Immunosuppressive Property of MSCs Mediated by Cell Surface Receptors

“…The biological activities of Gal-1 could promote not only repair and regeneration but also tumor metastasis in the tumor microenvironment (84,85). Besides, Gal-3 has been reported to have similar functions (86)(87)(88). However, compared to Gal-1, Gal-3 is more similar to Gal-9 in immunosuppression properties.…”

“…Direct coculture system (87) Complete medium supplemented with 10% FBS (88) Complete medium (90) Complete medium supplemented with 11% FBS (77) PD-L1 sPD-L1 PD-1 CD80 RGMb…”