Contributions
Abstract: P1182
Type: Poster
Abstract Category: Therapy - disease modifying - 27 Neuroprotection and Repair
Background: There is inconsistent neural stem cells (NSCs) differentiation and remyelination in MS lesions. Blockage of bone morphogenetic protein (BMP) is required for NSCs differentiation into neurons and oligodendrocytes, via downstream inhibition of pSMAD.
Objective: To examine the effect of BMP blockage in the MS model of relapsing experimental autoimmune encephalomyelitis (R-EAE).
Methods: BMP blockage done by neutralizing Abs or blocking small molecules (SMs) that were sorted by high throughput screening (HTS) with alkaline phosphatase production in ATDC5 cells as marker for BMP2 signaling and CellTiter-Glo as a marker for cell viability. Of 7,600 Maybridge SMs, 9 hits eventually demonstrated top IC50 values without toxic effect, named: SM1-SM9. R-EAE, induced by immunization with PLP139-151 in SJL mice were I.V treated with anti-hBMP-2/4, anti BMP-5, anti BMP-7 or isotype controls (IC) on day 9 post immunization. SM1-SM9 or vehicle were IP treated: day 9 -39 and BrdU in days: 9-18. In vitro neuronal phenotype was examined by MAP-2 expression in P19 cells.
Results: Combined anti-BMP-2/4, anti BMP-5, anti BMP-7 neutralizing Abs significantly ameliorated R-EAE and delayed 2nd relapse. Anti-BMP-2/4 alone significantly ameliorated R-EAE during whole experiment vs. anti-BMP-2/4,-5-7 altogether. Anti BMP-2/4 induced 2.9- and 3.5- fold of BrdU+DCX+ neuroblasts, reduced BrdU+GFAP+ NSCs in SGZ and SVZ vs. IC, and increased 5.0-fold of BrdU+O4+corpus callosum oligodendrocyte. No immunosuppressive effect was detected for anti-BMP-2/4 by measuring H&E brain infiltrates, and by splenocytes proliferation assay with anti-CD3 or PLP139-151. SM1 and SM9 significantly ameliorated R-EAE during 2nd, 3rd relapses vs. vehicle. Significant induction of BrdU+NeuN+ cells by SM1 and a 2.5- fold induction of BrdU+DCX+ cells in SVZ and 3.1 fold induction of BrdU+NeuN+ cells by SM9 were detected in R-EAE vs. the effect of the vehicle. SM1 and SM9 induced the neuronal MAP-2 expression in P19 neuroblastoma cell line. These SMs inhibited pSMAD expression studied by western blot in P19 cells. SM1 inhibit similarly the BMP4 signaling in ATDC5 cells while the inhibitory effect of SM9 on BMP4 was milder than on BMP2 signaling.
Conclusion: Systemic blockage of BMP-2/4 signaling have therapeutic potential to induce neurogenesis and oligodendrogenesis at the expense of astrogenesis in neuro-inflammatory diseases as MS, and to therefore to improve the repair of MS lesions.
Disclosure:
Funding: KAMIN program- Israeli economy and industry ministry.
Beyond that Dr Fainberg, Dr. Golan, Mr. Nen Hamou and Dr. Karni have nothing to disclose.
Abstract: P1182
Type: Poster
Abstract Category: Therapy - disease modifying - 27 Neuroprotection and Repair
Background: There is inconsistent neural stem cells (NSCs) differentiation and remyelination in MS lesions. Blockage of bone morphogenetic protein (BMP) is required for NSCs differentiation into neurons and oligodendrocytes, via downstream inhibition of pSMAD.
Objective: To examine the effect of BMP blockage in the MS model of relapsing experimental autoimmune encephalomyelitis (R-EAE).
Methods: BMP blockage done by neutralizing Abs or blocking small molecules (SMs) that were sorted by high throughput screening (HTS) with alkaline phosphatase production in ATDC5 cells as marker for BMP2 signaling and CellTiter-Glo as a marker for cell viability. Of 7,600 Maybridge SMs, 9 hits eventually demonstrated top IC50 values without toxic effect, named: SM1-SM9. R-EAE, induced by immunization with PLP139-151 in SJL mice were I.V treated with anti-hBMP-2/4, anti BMP-5, anti BMP-7 or isotype controls (IC) on day 9 post immunization. SM1-SM9 or vehicle were IP treated: day 9 -39 and BrdU in days: 9-18. In vitro neuronal phenotype was examined by MAP-2 expression in P19 cells.
Results: Combined anti-BMP-2/4, anti BMP-5, anti BMP-7 neutralizing Abs significantly ameliorated R-EAE and delayed 2nd relapse. Anti-BMP-2/4 alone significantly ameliorated R-EAE during whole experiment vs. anti-BMP-2/4,-5-7 altogether. Anti BMP-2/4 induced 2.9- and 3.5- fold of BrdU+DCX+ neuroblasts, reduced BrdU+GFAP+ NSCs in SGZ and SVZ vs. IC, and increased 5.0-fold of BrdU+O4+corpus callosum oligodendrocyte. No immunosuppressive effect was detected for anti-BMP-2/4 by measuring H&E brain infiltrates, and by splenocytes proliferation assay with anti-CD3 or PLP139-151. SM1 and SM9 significantly ameliorated R-EAE during 2nd, 3rd relapses vs. vehicle. Significant induction of BrdU+NeuN+ cells by SM1 and a 2.5- fold induction of BrdU+DCX+ cells in SVZ and 3.1 fold induction of BrdU+NeuN+ cells by SM9 were detected in R-EAE vs. the effect of the vehicle. SM1 and SM9 induced the neuronal MAP-2 expression in P19 neuroblastoma cell line. These SMs inhibited pSMAD expression studied by western blot in P19 cells. SM1 inhibit similarly the BMP4 signaling in ATDC5 cells while the inhibitory effect of SM9 on BMP4 was milder than on BMP2 signaling.
Conclusion: Systemic blockage of BMP-2/4 signaling have therapeutic potential to induce neurogenesis and oligodendrogenesis at the expense of astrogenesis in neuro-inflammatory diseases as MS, and to therefore to improve the repair of MS lesions.
Disclosure:
Funding: KAMIN program- Israeli economy and industry ministry.
Beyond that Dr Fainberg, Dr. Golan, Mr. Nen Hamou and Dr. Karni have nothing to disclose.