Contributions
Abstract: EP1501
Type: Poster Sessions
Abstract Category: Pathology and pathogenesis of MS - Repairing mechanisms
Background: Multiple sclerosis, a chronic inflammatory and degenerative disease of the CNS, is associated with damage of oligodendrocytes and myelin sheaths. Proliferation and differentiation of oligodendrocytes are influenced by fibroblast growth factors (FGFs) and mediated through FGF receptors (FGF1-3). To study the role of FGFR inhibition in myelin gene expression, we treated OLN-93 oligodendrocytes with AZD4547, a potent FGFR inhibitor, which is currently used in Phase II cancer trials and analyzed gene expressions of myelin markers and myelin inhibitors.
Methods: OLN-93 oligodendrocytes were incubated with AZD4547 (1 µM) and basic bFGF (25 ng). After 24 and 48 hours of incubation, proliferation of OLN-93 was measured using WST-1 proliferation assay and DAPI staining. Cytotoxicity was measured by LDH assay. Dead cell population was analyzed by 7AAD staining by FACS. Myelin inhibitors gene expression, FGF receptor (FGFR1) and the myelin marker CNPase was measured by RT-PCR. Phosphorylation of ERK1/2, AKT and p38 were analyzed by western blot.
Results: OLN93 cells treated with AZD4547 showed less proliferation (P < 0.01). Cytotoxicity was increased by AZD4547 (P < 0.05). There was no regulation of dead cell population by AZD4547. Phosphorylation of downstream molecules ERK, AKT and p38 was reduced by AZD4547. Gene expression of myelin inhibitors FGF2 (P < 0.05), SEMA3A (P < 0.05) and TGF-β (P < 0.01) was reduced, whereas the myelin marker CNPase was increased (P < 0.05).
Conclusions: Myelin inhibitors are known to play a key role in remyelination. FGFR inhibition by AZD4547 showed decreased proliferation and myelin inhibitors gene expression, whereas cytotoxicity was increased. Our data show that FGFR inhibition by AZD4547 mediates the myelin repair process through regulating myelin gene expression. The underlying downstream signaling mechanisms are downregulation of ERK and AKT phosphorylation by AZD4547.
Disclosure: Bischand Sharifi has nothing to disclose.
Ranjithkumar Rajendran has nothing to disclose.
Martin Berghoff has nothing to disclose.
Abstract: EP1501
Type: Poster Sessions
Abstract Category: Pathology and pathogenesis of MS - Repairing mechanisms
Background: Multiple sclerosis, a chronic inflammatory and degenerative disease of the CNS, is associated with damage of oligodendrocytes and myelin sheaths. Proliferation and differentiation of oligodendrocytes are influenced by fibroblast growth factors (FGFs) and mediated through FGF receptors (FGF1-3). To study the role of FGFR inhibition in myelin gene expression, we treated OLN-93 oligodendrocytes with AZD4547, a potent FGFR inhibitor, which is currently used in Phase II cancer trials and analyzed gene expressions of myelin markers and myelin inhibitors.
Methods: OLN-93 oligodendrocytes were incubated with AZD4547 (1 µM) and basic bFGF (25 ng). After 24 and 48 hours of incubation, proliferation of OLN-93 was measured using WST-1 proliferation assay and DAPI staining. Cytotoxicity was measured by LDH assay. Dead cell population was analyzed by 7AAD staining by FACS. Myelin inhibitors gene expression, FGF receptor (FGFR1) and the myelin marker CNPase was measured by RT-PCR. Phosphorylation of ERK1/2, AKT and p38 were analyzed by western blot.
Results: OLN93 cells treated with AZD4547 showed less proliferation (P < 0.01). Cytotoxicity was increased by AZD4547 (P < 0.05). There was no regulation of dead cell population by AZD4547. Phosphorylation of downstream molecules ERK, AKT and p38 was reduced by AZD4547. Gene expression of myelin inhibitors FGF2 (P < 0.05), SEMA3A (P < 0.05) and TGF-β (P < 0.01) was reduced, whereas the myelin marker CNPase was increased (P < 0.05).
Conclusions: Myelin inhibitors are known to play a key role in remyelination. FGFR inhibition by AZD4547 showed decreased proliferation and myelin inhibitors gene expression, whereas cytotoxicity was increased. Our data show that FGFR inhibition by AZD4547 mediates the myelin repair process through regulating myelin gene expression. The underlying downstream signaling mechanisms are downregulation of ERK and AKT phosphorylation by AZD4547.
Disclosure: Bischand Sharifi has nothing to disclose.
Ranjithkumar Rajendran has nothing to disclose.
Martin Berghoff has nothing to disclose.