Contributions
Abstract: EP1502
Type: Poster Sessions
Abstract Category: Pathology and pathogenesis of MS - Repairing mechanisms
Background: Fibroblast growth factors (FGFs) regulate the differentiation of immature oligodendrocytes into myelin-producing mature oligodendrocytes in the CNS. FGFR signalling is associated by activation of multiple cellular responses such as growth, proliferation, differentiation, and survival. In vitro and in vivo studies identified LINGO-1, TGF-β, SEMA3A and FGF2 as inhibitors of remyelination. BGJ398 is an orally bioavailable pan inhibitor of FGFRs with potential antiangiogenic and antineoplastic activities. In this study, we analysed the effect of BGJ398 on FGFR inhibition in oligodendrocytes and myelin related gene expression.
Methods: OLN93 oligodendrocytes were treated with BGJ398 (1 µM) and stimulated with basic FGF (20 ng) for 24 hours. Cell proliferation was analyzed by WST1 proliferation assay and DAPI staining. After treatments total mRNA was isolated from the cells and cDNA was synthesized. Myelin proteins (MBP, PLP, CNPase), myelin inhibitors (SEMA3A, FGF2, TGFβ) and NFG expression were analyzed by real time PCR. Quantitative expression of the genes was calculated by ΔΔCT method. MBP, TrkB and BDNF protein expression was analyzed by western blot and immunocytochemistry.
Results: Proliferation of OLN93 oligodendrocytes was reduced by BGJ398 (P < 0.05). FGFR1 (P < 0.05) and SEMA3A (P < 0.05) gene expression was downregulated by BGJ398. There was no regulation of FGF2 or TGFβ gene expression after BGJ398 treatment. CNPase (P < 0.05) and neural growth factor NGF gene expression (P < 0.05) were upregulated by BGJ398. There were no regulations in gene expression of MBP or PLP following BGJ398 treatment. MBP and BDNF protein expression was not regulated by BGJ398 or FGF. TrkB receptor expression was increased in BGJ398 treatment in western blot and immunocytochemistry (P < 0.05).
Conclusions: Our findings suggest that FGFR inhibition by BGJ398 in OLN93 oligodendrocytes enhances myelin gene expression and downregulates myelin inhibitor gene expression. The number of proliferating cells was less after BGJ398 treatment suggesting that FGFR inhibition supports the recovery of cells by enhancing myelin gene and growth factor expression. Application of BGJ398 is functional in the study of basic mechanisms involved in FGFR signaling in cells such as oligodendrocytes.
Disclosure: Vinothkumar Rajendran: nothing to disclose
Ranjithkumar Rajendran: nothing to disclose
Martin Berghoff: nothing to disclose
Abstract: EP1502
Type: Poster Sessions
Abstract Category: Pathology and pathogenesis of MS - Repairing mechanisms
Background: Fibroblast growth factors (FGFs) regulate the differentiation of immature oligodendrocytes into myelin-producing mature oligodendrocytes in the CNS. FGFR signalling is associated by activation of multiple cellular responses such as growth, proliferation, differentiation, and survival. In vitro and in vivo studies identified LINGO-1, TGF-β, SEMA3A and FGF2 as inhibitors of remyelination. BGJ398 is an orally bioavailable pan inhibitor of FGFRs with potential antiangiogenic and antineoplastic activities. In this study, we analysed the effect of BGJ398 on FGFR inhibition in oligodendrocytes and myelin related gene expression.
Methods: OLN93 oligodendrocytes were treated with BGJ398 (1 µM) and stimulated with basic FGF (20 ng) for 24 hours. Cell proliferation was analyzed by WST1 proliferation assay and DAPI staining. After treatments total mRNA was isolated from the cells and cDNA was synthesized. Myelin proteins (MBP, PLP, CNPase), myelin inhibitors (SEMA3A, FGF2, TGFβ) and NFG expression were analyzed by real time PCR. Quantitative expression of the genes was calculated by ΔΔCT method. MBP, TrkB and BDNF protein expression was analyzed by western blot and immunocytochemistry.
Results: Proliferation of OLN93 oligodendrocytes was reduced by BGJ398 (P < 0.05). FGFR1 (P < 0.05) and SEMA3A (P < 0.05) gene expression was downregulated by BGJ398. There was no regulation of FGF2 or TGFβ gene expression after BGJ398 treatment. CNPase (P < 0.05) and neural growth factor NGF gene expression (P < 0.05) were upregulated by BGJ398. There were no regulations in gene expression of MBP or PLP following BGJ398 treatment. MBP and BDNF protein expression was not regulated by BGJ398 or FGF. TrkB receptor expression was increased in BGJ398 treatment in western blot and immunocytochemistry (P < 0.05).
Conclusions: Our findings suggest that FGFR inhibition by BGJ398 in OLN93 oligodendrocytes enhances myelin gene expression and downregulates myelin inhibitor gene expression. The number of proliferating cells was less after BGJ398 treatment suggesting that FGFR inhibition supports the recovery of cells by enhancing myelin gene and growth factor expression. Application of BGJ398 is functional in the study of basic mechanisms involved in FGFR signaling in cells such as oligodendrocytes.
Disclosure: Vinothkumar Rajendran: nothing to disclose
Ranjithkumar Rajendran: nothing to disclose
Martin Berghoff: nothing to disclose