Contributions
Abstract: P639
Type: Poster
Abstract Category: Therapy - disease modifying - Immunomodulation/Immunosuppression
Background: Nrf2 (NF-E2-related factor 2) is a key regulator for cellular protection against oxidative stress. A ubiquitin ligase adaptor Keap1 (Kelch-like ECH-associated protein 1) negatively regulates Nrf2 by proteasome-mediated protein degradation. Multiple sclerosis (MS) is an autoimmune disease provoked by inflammation-mediated oxidative stress. The loss of Nrf2 exacerbates the development of experimental autoimmune encephalomyelitis (EAE) in mice. An Nrf2 inducer dimethyl fumarate (DMF) has been approved for the treatment of MS, and the Keap1-Nrf2 system is a promising target for treatment of MS. We have identified as a novel Nrf2 inducer TFM-735 by high throughput screening campaign. In this study, we examined the pharmacological profile of TFM-735, and evaluated therapeutic effects of TFM-735 on EAE in mice.
Methods: To examine the role of TFM-735 on Nrf2 stabilization, we performed immunoblot analysis by using thioglycollate-elicited mouse peritoneal macrophages from wild-type (WT) and Keap1C151S/C151S knock-in mice. The T-cell-mediated autoimmunity was evaluated by using the BioMAP® T cell Autoimmune Panel system (BioSeek). C57BL/6J background WT mice and ICR background highly sensitive inflammation monitoring hIL-6-BAC-Luc transgenic mice were immunized by myelin oligodendrocyte glycoprotein (MOG35-55) to induce EAE, and were orally administered with TFM-735 twice a day for 19 and 9 days, respectively.
Results: Nrf2 protein expression levels were increased by TFM-735 in the peritoneal macrophages from WT mice, but not in the macrophages from Keap1C151S/C151S knock-in mice. TFM-735 displayed an immune-modulative profile in BioMAP systems. TFM-735 repressed IL-17 secretion from PBMCs stimulated by anti-CD3 and anti-CD28 antibodies in a dose dependent manner. Administration of TFM-735 to mice increased mRNA expression levels of a representative Nrf2-target gene Nqo1 in brain and spleen, and decreased the clinical scores for EAE compared to administration of vehicle. The plasma IL-17 levels were also lowered in TFM-735-treated EAE mice compared to vehicle-treated EAE mice. In hIL-6-BAC-Luc transgenic mice, TFM-735 treatment reduced EAE-induced bioluminescence for IL-6 in central nervous system as well as clinical scores.
Conclusion: These results indicate that TFM-735 is a potent Nrf2 inducer that suppresses progression of EAE in mice. TFM-735 could be a promising therapeutic agent for MS.
Disclosure: All authors have nothing to disclose
Abstract: P639
Type: Poster
Abstract Category: Therapy - disease modifying - Immunomodulation/Immunosuppression
Background: Nrf2 (NF-E2-related factor 2) is a key regulator for cellular protection against oxidative stress. A ubiquitin ligase adaptor Keap1 (Kelch-like ECH-associated protein 1) negatively regulates Nrf2 by proteasome-mediated protein degradation. Multiple sclerosis (MS) is an autoimmune disease provoked by inflammation-mediated oxidative stress. The loss of Nrf2 exacerbates the development of experimental autoimmune encephalomyelitis (EAE) in mice. An Nrf2 inducer dimethyl fumarate (DMF) has been approved for the treatment of MS, and the Keap1-Nrf2 system is a promising target for treatment of MS. We have identified as a novel Nrf2 inducer TFM-735 by high throughput screening campaign. In this study, we examined the pharmacological profile of TFM-735, and evaluated therapeutic effects of TFM-735 on EAE in mice.
Methods: To examine the role of TFM-735 on Nrf2 stabilization, we performed immunoblot analysis by using thioglycollate-elicited mouse peritoneal macrophages from wild-type (WT) and Keap1C151S/C151S knock-in mice. The T-cell-mediated autoimmunity was evaluated by using the BioMAP® T cell Autoimmune Panel system (BioSeek). C57BL/6J background WT mice and ICR background highly sensitive inflammation monitoring hIL-6-BAC-Luc transgenic mice were immunized by myelin oligodendrocyte glycoprotein (MOG35-55) to induce EAE, and were orally administered with TFM-735 twice a day for 19 and 9 days, respectively.
Results: Nrf2 protein expression levels were increased by TFM-735 in the peritoneal macrophages from WT mice, but not in the macrophages from Keap1C151S/C151S knock-in mice. TFM-735 displayed an immune-modulative profile in BioMAP systems. TFM-735 repressed IL-17 secretion from PBMCs stimulated by anti-CD3 and anti-CD28 antibodies in a dose dependent manner. Administration of TFM-735 to mice increased mRNA expression levels of a representative Nrf2-target gene Nqo1 in brain and spleen, and decreased the clinical scores for EAE compared to administration of vehicle. The plasma IL-17 levels were also lowered in TFM-735-treated EAE mice compared to vehicle-treated EAE mice. In hIL-6-BAC-Luc transgenic mice, TFM-735 treatment reduced EAE-induced bioluminescence for IL-6 in central nervous system as well as clinical scores.
Conclusion: These results indicate that TFM-735 is a potent Nrf2 inducer that suppresses progression of EAE in mice. TFM-735 could be a promising therapeutic agent for MS.
Disclosure: All authors have nothing to disclose