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
A novel rheumatoid arthritis(RA)synovial fluid protein,Syntenin-1,and its receptor,Syndecan-1(SDC-1),are colocalized on RA synovial tissue endothelial cells and fibroblast-like synoviocytes(FLS).Syntenin-1 exacerbates...A novel rheumatoid arthritis(RA)synovial fluid protein,Syntenin-1,and its receptor,Syndecan-1(SDC-1),are colocalized on RA synovial tissue endothelial cells and fibroblast-like synoviocytes(FLS).Syntenin-1 exacerbates the inflammatory landscape of endothelial cells and RA FLS by upregulating transcription of IRF1/5/7/9,IL-1β,IL-6,and CCL2 through SDC-1 ligation and HIF1α,or mTOR activation.Mechanistically,Syntenin-1 orchestrates RA FLS and endothelial cell invasion via SDC-1 and/or mTOR signaling.In Syntenin-1 reprogrammed endothelial cells,the dynamic expression of metabolic intermediates coincides with escalated glycolysis along with unchanged oxidative factors,AMPK,PGC-1α,citrate,and inactive oxidative phosphorylation.Conversely,RA FLS rewired by Syntenin-1 displayed a modest glycolytic-ATP accompanied by a robust mitochondrial-ATP capacity.The enriched mitochondrial-ATP detected in Syntenin-1 reprogrammed RA FLS was coupled with mitochondrial fusion and fission recapitulated by escalated Mitofusin-2 and DRP1 expression.We found that VEGFR1/2 and Notch1 networks are responsible for the crosstalk between Syntenin-1 rewired endothelial cells and RA FLS,which are also represented in RA explants.Similar to RA explants,morphological and transcriptome studies authenticated the importance of VEGFR1/2,Notch1,RAPTOR,and HIF1αpathways in Syntenin-1 arthritic mice and their obstruction in SDC-1 deficient animals.Consistently,dysregulation of SDC-1,mTOR,and HIF1αnegated Syntenin-1 inflammatory phenotype in RA explants,while inhibition of HIF1αimpaired synovial angiogenic imprint amplified by Syntenin-1.In conclusion,since the current therapies are ineffective on Syntenin-1 and SDC-1 expression in RA synovial tissue and blood,targeting this pathway and its interconnected metabolic intermediates may provide a novel therapeutic strategy.展开更多
Flares of joint inflammation and resistance to currently available biologic therapeutics in rheumatoid arthritis(RA)patients could reflect activation of innate immune mechanisms.Herein,we show that a TLR7 GU-rich endo...Flares of joint inflammation and resistance to currently available biologic therapeutics in rheumatoid arthritis(RA)patients could reflect activation of innate immune mechanisms.Herein,we show that a TLR7 GU-rich endogenous ligand,miR-Let7b,potentiates synovitis by amplifying RA monocyte and fibroblast(FLS)trafficking.miR-Let7b ligation to TLR7 in macrophages(MΦs)and FLSs expanded the synovial inflammatory response.Moreover,secretion of M1 monokines triggered by miR-Let7b enhanced Th1/Th17 cell differentiation.We showed that IRAK4 inhibitor(i)therapy attenuated RA disease activity by blocking TLR7-induced M1 MΦor FLS activation,as well as monokine-modulated Th1/Th17 cell polarization.IRAK4i therapy also disrupted RA osteoclastogenesis,which was amplified by miR-Let7b ligation to joint myeloid TLR7.Hence,the effectiveness of IRAK4i was compared with that of a TNF inhibitor(i)or anti-IL-6R treatment in collagen-induced arthritis(CIA)and miR-Let7b-mediated arthritis.We found that TNF or IL-6R blocking therapies mitigated CIA by reducing the infiltration of joint F480+iNOS+MΦs,the expression of certain monokines,and Th1 cell differentiation.Unexpectedly,these biologic therapies were unable to alleviate miR-Let7b-induced arthritis.The superior efficacy of IRAK4i over anti-TNF or anti-IL-6R therapy in miR-Let7b-induced arthritis or CIA was due to the ability of IRAK4i therapy to restrain the migration of joint F480+iNOS+MΦs,vimentin+fibroblasts,and CD3+T cells,in addition to negating the expression of a wide range of monokines,including IL-12,MIP2,and IRF5 and Th1/Th17 lymphokines.In conclusion,IRAK4i therapy may provide a promising strategy for RA therapy by disconnecting critical links between inflammatory joint cells.展开更多
IL-34 is a novel biomarker for rheumatoid arthritis(RA),as its serum levels are linked to C-reactive protein,erythrocyte sedimentation rate,rheumatoid factor(RF),IL-6 and radiographic progression[1,2].IL-34 reprograms...IL-34 is a novel biomarker for rheumatoid arthritis(RA),as its serum levels are linked to C-reactive protein,erythrocyte sedimentation rate,rheumatoid factor(RF),IL-6 and radiographic progression[1,2].IL-34 reprograms naïve circulating M0 cells into glycolytic(CD14^(+)CD86^(+)GLUT1^(high))M34 macrophages(MΦs)by binding to its pathogenic receptor,SDC-1,which controls the activation of its coreceptor/M-CSFR[3].Glycolytic M34 MΦs are primed to differentiate into osteoclasts through SDC-1 ligation[3].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.
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 AI167155,NIH R41 AI147697the Innovative Research Award from the Rheumatology Research Foundation(RRF,no number assigned).
文摘A novel rheumatoid arthritis(RA)synovial fluid protein,Syntenin-1,and its receptor,Syndecan-1(SDC-1),are colocalized on RA synovial tissue endothelial cells and fibroblast-like synoviocytes(FLS).Syntenin-1 exacerbates the inflammatory landscape of endothelial cells and RA FLS by upregulating transcription of IRF1/5/7/9,IL-1β,IL-6,and CCL2 through SDC-1 ligation and HIF1α,or mTOR activation.Mechanistically,Syntenin-1 orchestrates RA FLS and endothelial cell invasion via SDC-1 and/or mTOR signaling.In Syntenin-1 reprogrammed endothelial cells,the dynamic expression of metabolic intermediates coincides with escalated glycolysis along with unchanged oxidative factors,AMPK,PGC-1α,citrate,and inactive oxidative phosphorylation.Conversely,RA FLS rewired by Syntenin-1 displayed a modest glycolytic-ATP accompanied by a robust mitochondrial-ATP capacity.The enriched mitochondrial-ATP detected in Syntenin-1 reprogrammed RA FLS was coupled with mitochondrial fusion and fission recapitulated by escalated Mitofusin-2 and DRP1 expression.We found that VEGFR1/2 and Notch1 networks are responsible for the crosstalk between Syntenin-1 rewired endothelial cells and RA FLS,which are also represented in RA explants.Similar to RA explants,morphological and transcriptome studies authenticated the importance of VEGFR1/2,Notch1,RAPTOR,and HIF1αpathways in Syntenin-1 arthritic mice and their obstruction in SDC-1 deficient animals.Consistently,dysregulation of SDC-1,mTOR,and HIF1αnegated Syntenin-1 inflammatory phenotype in RA explants,while inhibition of HIF1αimpaired synovial angiogenic imprint amplified by Syntenin-1.In conclusion,since the current therapies are ineffective on Syntenin-1 and SDC-1 expression in RA synovial tissue and blood,targeting this pathway and its interconnected metabolic intermediates may provide a novel therapeutic strategy.
基金This work was supported in part by awards from the Department of Veteran’s Affairs MERIT Award(1I01BX002286)the National Institutes of Health(AR056099 and AR065778)+2 种基金the National Psoriasis Foundation(NPF)the Pfizer Investigator Initiated Research(IIR)Programthe Chicago Biomedical Consortium(CBC)Accelerator Award.
文摘Flares of joint inflammation and resistance to currently available biologic therapeutics in rheumatoid arthritis(RA)patients could reflect activation of innate immune mechanisms.Herein,we show that a TLR7 GU-rich endogenous ligand,miR-Let7b,potentiates synovitis by amplifying RA monocyte and fibroblast(FLS)trafficking.miR-Let7b ligation to TLR7 in macrophages(MΦs)and FLSs expanded the synovial inflammatory response.Moreover,secretion of M1 monokines triggered by miR-Let7b enhanced Th1/Th17 cell differentiation.We showed that IRAK4 inhibitor(i)therapy attenuated RA disease activity by blocking TLR7-induced M1 MΦor FLS activation,as well as monokine-modulated Th1/Th17 cell polarization.IRAK4i therapy also disrupted RA osteoclastogenesis,which was amplified by miR-Let7b ligation to joint myeloid TLR7.Hence,the effectiveness of IRAK4i was compared with that of a TNF inhibitor(i)or anti-IL-6R treatment in collagen-induced arthritis(CIA)and miR-Let7b-mediated arthritis.We found that TNF or IL-6R blocking therapies mitigated CIA by reducing the infiltration of joint F480+iNOS+MΦs,the expression of certain monokines,and Th1 cell differentiation.Unexpectedly,these biologic therapies were unable to alleviate miR-Let7b-induced arthritis.The superior efficacy of IRAK4i over anti-TNF or anti-IL-6R therapy in miR-Let7b-induced arthritis or CIA was due to the ability of IRAK4i therapy to restrain the migration of joint F480+iNOS+MΦs,vimentin+fibroblasts,and CD3+T cells,in addition to negating the expression of a wide range of monokines,including IL-12,MIP2,and IRF5 and Th1/Th17 lymphokines.In conclusion,IRAK4i therapy may provide a promising strategy for RA therapy by disconnecting critical links between inflammatory joint cells.
基金This work was supported in part by awards from the Department of Veteran’s Affairs MERIT Award BX002286the Innovative Research Award from the Rheumatology Research Foundation(RRF)+3 种基金the National Institutes of Health NIH AI147697the National Psoriasis Foundation(NPF)the Pfizer Investigator-Initiated Research(IIR)Programthe Chicago Biomedical Consortium(CBC)Accelerator Award.
文摘IL-34 is a novel biomarker for rheumatoid arthritis(RA),as its serum levels are linked to C-reactive protein,erythrocyte sedimentation rate,rheumatoid factor(RF),IL-6 and radiographic progression[1,2].IL-34 reprograms naïve circulating M0 cells into glycolytic(CD14^(+)CD86^(+)GLUT1^(high))M34 macrophages(MΦs)by binding to its pathogenic receptor,SDC-1,which controls the activation of its coreceptor/M-CSFR[3].Glycolytic M34 MΦs are primed to differentiate into osteoclasts through SDC-1 ligation[3].
