Background:Smooth muscle cells(SMCs)exhibit remarkable plasticity,undergoing extensive phenotypic switching to gener-ate a highly heterogeneous population within atherosclerotic plaques.While recent studies have highl...Background:Smooth muscle cells(SMCs)exhibit remarkable plasticity,undergoing extensive phenotypic switching to gener-ate a highly heterogeneous population within atherosclerotic plaques.While recent studies have highlighted the contribution of SMC-derived macrophage-like cells to plaque inflammation,the specific molecular drivers governing the transition to these pathogenic states remain poorly understood.Methods:Here,we re-analyzed single-cell RNA sequencing data from lineage-traced mice to dissect SMC heterogeneity during atherogenesis.Trajectory analysis revealed that SMCs transdifferentiate into a distinct pro-inflammatory macrophage-like subpopulation(macrophage 4)via an intermediate"stem-endothelial-monocyte"cell state.Integrated gene regulatory net-work inference and in silico perturbation modeling identified interferon regulatory factor 7(IRF7)as a master transcriptional regulator orchestrating this specific pathogenic transition.Results:Clinically,IRF7 expression was significantly upregulated in unstable and advanced human atherosclerotic plaques,correlating strongly with inflammatory macrophage burden.In vivo,ApoE^(−/−)mice challenged with a high-fat diet exhibited robust upregulation of IRF7 in aortic plaques,which co-localized with macrophage markers.Crucially,SMC-specific knock-down of Irf7 using an AAV-SM22α-shIRF7 vector significantly attenuated atherosclerotic plaque progression,reduced necrotic core formation,and enhanced fibrous cap stability.Mechanistically,Irf7 silencing preserved the contractile SMC phenotype and inhibited the accumulation of pro-inflammatory SMC-derived macrophage-like cells within the lesion.Conclusions:These findings identify IRF7 as a critical checkpoint in maladaptive SMC phenotype switching.We demonstrate that IRF7 drives the transdifferentiation of SMCs into a pro-inflammatory macrophage-like state,thereby fueling plaque insta-bility.Consequently,therapeutic strategies capable of inhibiting IRF7-mediated SMC plasticity may prove effective in stabiliz-ing vulnerable atherosclerotic plaques.展开更多
p53 is mutated in half of cancer cases.However,no p53-targeting drugs have been approved.Here,we reposition decitabine for triple-negative breast cancer(TNBC),a subtype with frequent p53 mutations and extremely poor p...p53 is mutated in half of cancer cases.However,no p53-targeting drugs have been approved.Here,we reposition decitabine for triple-negative breast cancer(TNBC),a subtype with frequent p53 mutations and extremely poor prognosis.In a retrospective study on tissue microarrays with 132 TNBC cases,DNMT1 overexpression was associated with p53 mutations(P=0.037)and poor overall survival(OS)(P=0.010).In a prospective DEciTabinE and Carboplatin in TNBC(DETECT)trial(NCT03295552),decitabine with carboplatin produced an objective response rate(ORR)of 42%in 12 patients with stage IV TNBC.Among the 9 trialed patients with available TP53 sequencing results,the 6 patients with p53 mutations had higher ORR(3/6 vs.0/3)and better OS(16.0 vs.4.0 months)than the patients with wild-type p53.In a mechanistic study,isogenic TNBC cell lines harboring DETECT-derived p53 mutations exhibited higher DNMT1 expression and decitabine sensitivity than the cell line with wild-type p53.In the DETECT trial,decitabine induced strong immune responses featuring the striking upregulation of the innate immune player IRF7 in the p53-mutated TNBC cell line(upregulation by 16-fold)and the most responsive patient with TNBC.Our integrative studies reveal the potential of repurposing decitabine for the treatment of p53-mutated TNBC and suggest IRF7 as a potential biomarker for decitabine-based treatments.展开更多
The peripheral nervous system has the potential to regenerate after nerve injury owing to the intrinsic regrowth ability of neurons and the permissive microenvironment.The regenerative process involves numerous gene e...The peripheral nervous system has the potential to regenerate after nerve injury owing to the intrinsic regrowth ability of neurons and the permissive microenvironment.The regenerative process involves numerous gene expression changes,in which transcription factors play a critical role.Previously,we profiled dysregulated genes in dorsal root ganglion neurons at different time points（0,3 and 9 hours,and 1,4 and 7 days） after sciatic nerve injury in rats by RNA sequencing.In the present study,we investigated differentially expressed transcription factors following nerve injury,and we identified enriched molecular and cellular functions of these transcription factors by Ingenuity Pathway Analysis.This analysis revealed the dynamic changes in the expression of transcription factors involved in cell death at different time points following sciatic nerve injury.In addition,we constructed regulatory networks of the differentially expressed transcription factors in cell death and identified some key transcription factors（such as STAT1,JUN,MYC and IRF7）.We confirmed the changes in expression of some key transcription factors（STAT1 and IRF7） by quantitative reverse transcription-polymerase chain reaction.Collectively,our analyses provide a global overview of transcription factor changes in dorsal root ganglia after sciatic nerve injury and offer insight into the regulatory transcription factor networks involved in cell death.展开更多
基金supported by the National Natural Science Foun-dation of China(grant No.32171387).
文摘Background:Smooth muscle cells(SMCs)exhibit remarkable plasticity,undergoing extensive phenotypic switching to gener-ate a highly heterogeneous population within atherosclerotic plaques.While recent studies have highlighted the contribution of SMC-derived macrophage-like cells to plaque inflammation,the specific molecular drivers governing the transition to these pathogenic states remain poorly understood.Methods:Here,we re-analyzed single-cell RNA sequencing data from lineage-traced mice to dissect SMC heterogeneity during atherogenesis.Trajectory analysis revealed that SMCs transdifferentiate into a distinct pro-inflammatory macrophage-like subpopulation(macrophage 4)via an intermediate"stem-endothelial-monocyte"cell state.Integrated gene regulatory net-work inference and in silico perturbation modeling identified interferon regulatory factor 7(IRF7)as a master transcriptional regulator orchestrating this specific pathogenic transition.Results:Clinically,IRF7 expression was significantly upregulated in unstable and advanced human atherosclerotic plaques,correlating strongly with inflammatory macrophage burden.In vivo,ApoE^(−/−)mice challenged with a high-fat diet exhibited robust upregulation of IRF7 in aortic plaques,which co-localized with macrophage markers.Crucially,SMC-specific knock-down of Irf7 using an AAV-SM22α-shIRF7 vector significantly attenuated atherosclerotic plaque progression,reduced necrotic core formation,and enhanced fibrous cap stability.Mechanistically,Irf7 silencing preserved the contractile SMC phenotype and inhibited the accumulation of pro-inflammatory SMC-derived macrophage-like cells within the lesion.Conclusions:These findings identify IRF7 as a critical checkpoint in maladaptive SMC phenotype switching.We demonstrate that IRF7 drives the transdifferentiation of SMCs into a pro-inflammatory macrophage-like state,thereby fueling plaque insta-bility.Consequently,therapeutic strategies capable of inhibiting IRF7-mediated SMC plasticity may prove effective in stabiliz-ing vulnerable atherosclerotic plaques.
基金supported by the National Natural Science Foundation of China(No.82130075 to Min Lu,No.82073292 to Min Lu,No.81772797 to Xiaosong Chen,No.82072937 to Xiaosong Chen,No.82072897 to Kunwei Shen,No.82002773 to Zheng Wang,No.81900157 to Ying Liang)SJTU Transmed Awards Research(to Min Lu),Shanghai Municipal Education Commission-Gaofeng Clinical Medicine(No.828318 to Min Lu and No.20172007 to Xiaosong Chen)+4 种基金Shanghai Excellent Youth Academic Leader(No.20XD1422700 to Min Lu)Program of Shanghai Science and Technology Committee(No.21S11900100 to Min Lu)Dawn Program of Shanghai Education Commission(No.21SG18 to Min Lu)Samuel Waxman Cancer Research Foundation(to Min Lu)Foundation of National Facility for Translational Medicine(Shanghai)(No.NRCTM(SH)-2021-08).
文摘p53 is mutated in half of cancer cases.However,no p53-targeting drugs have been approved.Here,we reposition decitabine for triple-negative breast cancer(TNBC),a subtype with frequent p53 mutations and extremely poor prognosis.In a retrospective study on tissue microarrays with 132 TNBC cases,DNMT1 overexpression was associated with p53 mutations(P=0.037)and poor overall survival(OS)(P=0.010).In a prospective DEciTabinE and Carboplatin in TNBC(DETECT)trial(NCT03295552),decitabine with carboplatin produced an objective response rate(ORR)of 42%in 12 patients with stage IV TNBC.Among the 9 trialed patients with available TP53 sequencing results,the 6 patients with p53 mutations had higher ORR(3/6 vs.0/3)and better OS(16.0 vs.4.0 months)than the patients with wild-type p53.In a mechanistic study,isogenic TNBC cell lines harboring DETECT-derived p53 mutations exhibited higher DNMT1 expression and decitabine sensitivity than the cell line with wild-type p53.In the DETECT trial,decitabine induced strong immune responses featuring the striking upregulation of the innate immune player IRF7 in the p53-mutated TNBC cell line(upregulation by 16-fold)and the most responsive patient with TNBC.Our integrative studies reveal the potential of repurposing decitabine for the treatment of p53-mutated TNBC and suggest IRF7 as a potential biomarker for decitabine-based treatments.
基金supported by the National Natural Science Foundation of China,No.31500823the Natural Science Foundation of Jiangsu Province of China,No.BK20150403the Natural Science Fund for Colleges and Universities in Jiangsu Province of China,No.16KJB180024
文摘The peripheral nervous system has the potential to regenerate after nerve injury owing to the intrinsic regrowth ability of neurons and the permissive microenvironment.The regenerative process involves numerous gene expression changes,in which transcription factors play a critical role.Previously,we profiled dysregulated genes in dorsal root ganglion neurons at different time points（0,3 and 9 hours,and 1,4 and 7 days） after sciatic nerve injury in rats by RNA sequencing.In the present study,we investigated differentially expressed transcription factors following nerve injury,and we identified enriched molecular and cellular functions of these transcription factors by Ingenuity Pathway Analysis.This analysis revealed the dynamic changes in the expression of transcription factors involved in cell death at different time points following sciatic nerve injury.In addition,we constructed regulatory networks of the differentially expressed transcription factors in cell death and identified some key transcription factors（such as STAT1,JUN,MYC and IRF7）.We confirmed the changes in expression of some key transcription factors（STAT1 and IRF7） by quantitative reverse transcription-polymerase chain reaction.Collectively,our analyses provide a global overview of transcription factor changes in dorsal root ganglia after sciatic nerve injury and offer insight into the regulatory transcription factor networks involved in cell death.
