Rheumatoid arthritis(RA)exhibits heterogeneous endotypes,complicating treatment strategies.GM-CSF and GM-CSFRα are enriched in RA synovial CD68⁺macrophages(MΦs),and are implicated in acute and chronic disease stages...Rheumatoid arthritis(RA)exhibits heterogeneous endotypes,complicating treatment strategies.GM-CSF and GM-CSFRα are enriched in RA synovial CD68⁺macrophages(MΦs),and are implicated in acute and chronic disease stages.Since anti-TNFi and anti-IL6R therapies did not effectively suppress GM-CSF/GM-CSFRα expression or the GM-CSF-associated landscape,we explored alternative therapeutic strategies to target GM-CSF function using RA blood,synovial tissues,and preclinical models.We demonstrate that GM-CSF-MΦs reprogrammed in RA blood and synovial tissue share a distinct IL1β*S100A*HIF1*IL10^(10)NFIL3/6^(10)expression profile,manifested by mitochondrial oxidative stress and fragmentation.To correct the metabolic imbalance of GM-CSFMΦs,cells were treated with a complex I inhibitor(i)or a glucose uptake blocker.Complex Ii did not broadly alter the inflammatory or metabolic networks or affect the mitochondrial dynamics remodeled by GM-CSF-MΦs.While the glucose uptake inhibitor(HK2i)reduced glycolysis-derived ATP,it had limited efficacy in restricting the inflammatory signature or restoring TCA enzymes in GMCSF-MΦs.In contrast,tofacitinib achieved broad-spectrum effects by downregulating GM-CSFRαexpression and inhibiting STAT5 signaling.Moreover,tofacitinib redirected RA blood and synovial IL1β*S100A*HIF1*IL10^(10)NFIL3/6^(10)MΦs into a regulatory phenotype,reversing oxidative stress and mitochondrial fragmentation.In preclinical models,local GM-CSF overexpression induced MΦ-directed joint inflammation and metabolic dysregulation.Consistently,Tofacitinib reversed GM-CSF-differentiated murine IL1β*HBEGF*HIF1*MΦs by impeding STAT5 signaling,correcting metabolic dysregulation,and repairing mitochondrial fragmentation.In conclusion,anti-TNFi,anti-IL6R,and metabolic-targeted therapies were largely ineffective in modifying GM-CSFMΦ pathology.Conversely,tofacitinib deactivation of STAT5 attenuates GM-CSF-MΦ-triggered inflammation and mitochondrial malfunction by restoring regulatory markers and rebalancing oxidative phosphorylation in RA specimens and/or preclinical models.展开更多
基金supported in part by awards from the Department of Veteran’s Affairs MERIT Award BX002286,CX002565,IK6BX006474the National Institutes of Health NIH R01 AI167155the Innovative Research Award from the Rheumatology Research Foundation(RRF,no number assigned).
文摘Rheumatoid arthritis(RA)exhibits heterogeneous endotypes,complicating treatment strategies.GM-CSF and GM-CSFRα are enriched in RA synovial CD68⁺macrophages(MΦs),and are implicated in acute and chronic disease stages.Since anti-TNFi and anti-IL6R therapies did not effectively suppress GM-CSF/GM-CSFRα expression or the GM-CSF-associated landscape,we explored alternative therapeutic strategies to target GM-CSF function using RA blood,synovial tissues,and preclinical models.We demonstrate that GM-CSF-MΦs reprogrammed in RA blood and synovial tissue share a distinct IL1β*S100A*HIF1*IL10^(10)NFIL3/6^(10)expression profile,manifested by mitochondrial oxidative stress and fragmentation.To correct the metabolic imbalance of GM-CSFMΦs,cells were treated with a complex I inhibitor(i)or a glucose uptake blocker.Complex Ii did not broadly alter the inflammatory or metabolic networks or affect the mitochondrial dynamics remodeled by GM-CSF-MΦs.While the glucose uptake inhibitor(HK2i)reduced glycolysis-derived ATP,it had limited efficacy in restricting the inflammatory signature or restoring TCA enzymes in GMCSF-MΦs.In contrast,tofacitinib achieved broad-spectrum effects by downregulating GM-CSFRαexpression and inhibiting STAT5 signaling.Moreover,tofacitinib redirected RA blood and synovial IL1β*S100A*HIF1*IL10^(10)NFIL3/6^(10)MΦs into a regulatory phenotype,reversing oxidative stress and mitochondrial fragmentation.In preclinical models,local GM-CSF overexpression induced MΦ-directed joint inflammation and metabolic dysregulation.Consistently,Tofacitinib reversed GM-CSF-differentiated murine IL1β*HBEGF*HIF1*MΦs by impeding STAT5 signaling,correcting metabolic dysregulation,and repairing mitochondrial fragmentation.In conclusion,anti-TNFi,anti-IL6R,and metabolic-targeted therapies were largely ineffective in modifying GM-CSFMΦ pathology.Conversely,tofacitinib deactivation of STAT5 attenuates GM-CSF-MΦ-triggered inflammation and mitochondrial malfunction by restoring regulatory markers and rebalancing oxidative phosphorylation in RA specimens and/or preclinical models.
