Background:Immune checkpoint inhibitors(ICIs)targeting programmed cell death protein 1(PD-1)and programmed death-ligand 1(PD-L1)have shown a moderate response in colorectal cancer(CRC)with deficient mismatch repair(dM...Background:Immune checkpoint inhibitors(ICIs)targeting programmed cell death protein 1(PD-1)and programmed death-ligand 1(PD-L1)have shown a moderate response in colorectal cancer(CRC)with deficient mismatch repair(dMMR)functions and poor response in patients with proficientMMR(pMMR).pMMRtumors are generally immunogenically“cold”,emphasizing combination strategies to turn the“cold”tumor“hot”to enhance the efficacy of ICIs.ATR inhibitors(ATRi)have been proven to cooperate with radiation to promote antitumor immunity,but it is unclear whether ATRi could facilitate the efficacy of IR and ICI combinations in CRCs.This study aimed to investigate the efficacy of combining ATRi,irradiation(IR),and anti-PD-L1 antibodies in CRC mouse models with different microsatellite statuses.Methods:The efficacy of combining ATRi,IR,and anti-PD-L1 antibodies was evaluated in CRC tumors.The tumor microenvironment and transcriptome signatures were investigated under different treatment regimens.The mechanisms were explored via cell viability assay,flow cytometry,immunofluorescence,immunoblotting,co-immunoprecipitation,and real-time quantitative PCR in multiple murine and human CRC cell lines.Results:Combining ATRi berzosertib and IR enhanced CD8+T cell infiltration and enhanced the efficacy of anti-PD-L1 therapy in mouse CRC models with different microsatellite statuses.The mechanistic study demonstrated that IR+ATRi could activate both the canonical cGAS-STING-pTBK1/pIRF3 axis by increasing cytosolic double-stranded DNA levels and the non-canonical STING signaling by attenuating SHP1-mediated inhibition of the TRAF6-STINGp65 axis,via promoting SUMOylation of SHP1 at lysine 127.By boosting the STING signaling,IR+ATRi induced type I interferon-related gene expression and strong innate immune activation and reinvigorated the cold tumor microenvironment,thus facilitating immunotherapy.Conclusions:The combination of ATRi and IR could facilitate anti-PD-L1 therapy by promoting STING signaling in CRC models with different microsatellite statuses.The new combination strategy raised by our study isworth investigating in the management of CRC.展开更多
[目的]探究SHP-1与NF-κB对深静脉血栓血清炎症因子水平的影响。[方法]通过免疫组化和蛋白免疫印迹实验分析下肢静脉血管组织中的SHP-1表达水平;将深静脉血栓患者的外周血单个核细胞分为3组:对照组,pcDNA NC组和pcDNA SHP-1组。通过酶...[目的]探究SHP-1与NF-κB对深静脉血栓血清炎症因子水平的影响。[方法]通过免疫组化和蛋白免疫印迹实验分析下肢静脉血管组织中的SHP-1表达水平;将深静脉血栓患者的外周血单个核细胞分为3组:对照组,pcDNA NC组和pcDNA SHP-1组。通过酶联免疫吸附试验分析外周血单个核细胞分泌的IL-1β、IL-6、IFN-γ和TNF-α的浓度;通过蛋白免疫印迹方法检测不同实验组的外周血单个核细胞中SHP-1以及NF-κB p65的蛋白水平。[结果]免疫组化实验结果显示,SHP-1在深静脉血栓患者的血管组织中表达降低(P<0.05),NF-κB p65在深静脉血栓患者的血管组织中血管组织中表达增加(0.21±0.01 vs 0.82±0.05,P<0.05);酶联免疫吸附试验结果显示,pcDNA SHP-1组的炎症因子IL-1β、IL-6、IFN-γ和TNF-α的释放水平下降(P<0.05);蛋白免疫印迹实验结果显示,pcDNA SHP-1组的SHP-1蛋白表达水平增加(0.19±0.02 vs 0.22±0.03 vs 0.82±0.06,P<0.05),而NF-κB p65蛋白表达水平降低(0.71±0.03 vs 0.73±0.05 vs 0.25±0.02,P<0.05)。[结论]在深静脉血栓患者体内,SHP-1表达下降能够导致NF-κB p65蛋白表达增加,进而促进外周血单个核细胞释放IL-1β、IL-6、IFN-γ和TNF-α等炎症因子,加重炎症反应程度,SHP-1是深静脉血栓治疗的潜在靶点。展开更多
Atherosclerotic cardiovascular disease is the leading cause of mortality in the world.A driving feature of atherosclerotic plaque formation is dysfunctional efferocytosis.Because the“don’t eat me”molecule CD47 is u...Atherosclerotic cardiovascular disease is the leading cause of mortality in the world.A driving feature of atherosclerotic plaque formation is dysfunctional efferocytosis.Because the“don’t eat me”molecule CD47 is upregulated in atherosclerotic plaque cores,CD47-blocking strategies can stimulate the efferocytic clearance of apoptotic cells and thereby help prevent the progression of plaque buildup.However,these therapies are generally costly and,in clinical and murine trials,they have resulted in side effects including anemia and reticulocytosis.Here,we developed and characterized an intracellular phagocytosis-stimulating treatment in the CD47-SIRPαpathway.We loaded a novel monocyte/macrophage-selective nanoparticle carrier system with a small molecule enzymatic inhibitor that is released in a pH-dependent manner to stimulate macrophage efferocytosis of apoptotic cell debris via the CD47-SIRPαsignaling pathway.We demonstrated that single-walled carbon nanotubes(SWNTs)can selectively deliver tyrosine phosphatase inhibitor 1(TPI)intracellularly to macrophages,which potently stimulates efferocytosis,and chemically characterized the nanocarrier.Thus,SWNT-delivered TPI can stimulate macrophage efferocytosis,with the potential to reduce or prevent atherosclerotic disease.展开更多
基金Innovative Capacity Building Project of the Hubei Engineering Research Center for Radiotherapy and Radiation Protection of Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Grant/Award Number:2018-420114-35-03-071705State Key Program of National Natural Science of China,Grant/Award Number:82130092National Natural Science Foundation of China,Grant/Award Numbers:81372664,81902619。
文摘Background:Immune checkpoint inhibitors(ICIs)targeting programmed cell death protein 1(PD-1)and programmed death-ligand 1(PD-L1)have shown a moderate response in colorectal cancer(CRC)with deficient mismatch repair(dMMR)functions and poor response in patients with proficientMMR(pMMR).pMMRtumors are generally immunogenically“cold”,emphasizing combination strategies to turn the“cold”tumor“hot”to enhance the efficacy of ICIs.ATR inhibitors(ATRi)have been proven to cooperate with radiation to promote antitumor immunity,but it is unclear whether ATRi could facilitate the efficacy of IR and ICI combinations in CRCs.This study aimed to investigate the efficacy of combining ATRi,irradiation(IR),and anti-PD-L1 antibodies in CRC mouse models with different microsatellite statuses.Methods:The efficacy of combining ATRi,IR,and anti-PD-L1 antibodies was evaluated in CRC tumors.The tumor microenvironment and transcriptome signatures were investigated under different treatment regimens.The mechanisms were explored via cell viability assay,flow cytometry,immunofluorescence,immunoblotting,co-immunoprecipitation,and real-time quantitative PCR in multiple murine and human CRC cell lines.Results:Combining ATRi berzosertib and IR enhanced CD8+T cell infiltration and enhanced the efficacy of anti-PD-L1 therapy in mouse CRC models with different microsatellite statuses.The mechanistic study demonstrated that IR+ATRi could activate both the canonical cGAS-STING-pTBK1/pIRF3 axis by increasing cytosolic double-stranded DNA levels and the non-canonical STING signaling by attenuating SHP1-mediated inhibition of the TRAF6-STINGp65 axis,via promoting SUMOylation of SHP1 at lysine 127.By boosting the STING signaling,IR+ATRi induced type I interferon-related gene expression and strong innate immune activation and reinvigorated the cold tumor microenvironment,thus facilitating immunotherapy.Conclusions:The combination of ATRi and IR could facilitate anti-PD-L1 therapy by promoting STING signaling in CRC models with different microsatellite statuses.The new combination strategy raised by our study isworth investigating in the management of CRC.
文摘[目的]探究SHP-1与NF-κB对深静脉血栓血清炎症因子水平的影响。[方法]通过免疫组化和蛋白免疫印迹实验分析下肢静脉血管组织中的SHP-1表达水平;将深静脉血栓患者的外周血单个核细胞分为3组:对照组,pcDNA NC组和pcDNA SHP-1组。通过酶联免疫吸附试验分析外周血单个核细胞分泌的IL-1β、IL-6、IFN-γ和TNF-α的浓度;通过蛋白免疫印迹方法检测不同实验组的外周血单个核细胞中SHP-1以及NF-κB p65的蛋白水平。[结果]免疫组化实验结果显示,SHP-1在深静脉血栓患者的血管组织中表达降低(P<0.05),NF-κB p65在深静脉血栓患者的血管组织中血管组织中表达增加(0.21±0.01 vs 0.82±0.05,P<0.05);酶联免疫吸附试验结果显示,pcDNA SHP-1组的炎症因子IL-1β、IL-6、IFN-γ和TNF-α的释放水平下降(P<0.05);蛋白免疫印迹实验结果显示,pcDNA SHP-1组的SHP-1蛋白表达水平增加(0.19±0.02 vs 0.22±0.03 vs 0.82±0.06,P<0.05),而NF-κB p65蛋白表达水平降低(0.71±0.03 vs 0.73±0.05 vs 0.25±0.02,P<0.05)。[结论]在深静脉血栓患者体内,SHP-1表达下降能够导致NF-κB p65蛋白表达增加,进而促进外周血单个核细胞释放IL-1β、IL-6、IFN-γ和TNF-α等炎症因子,加重炎症反应程度,SHP-1是深静脉血栓治疗的潜在靶点。
基金This study was funded by an AHA Transformational Project grant(No.18TPA34230113),NIH R01 CA244491,and Falk Catalyst grant.
文摘Atherosclerotic cardiovascular disease is the leading cause of mortality in the world.A driving feature of atherosclerotic plaque formation is dysfunctional efferocytosis.Because the“don’t eat me”molecule CD47 is upregulated in atherosclerotic plaque cores,CD47-blocking strategies can stimulate the efferocytic clearance of apoptotic cells and thereby help prevent the progression of plaque buildup.However,these therapies are generally costly and,in clinical and murine trials,they have resulted in side effects including anemia and reticulocytosis.Here,we developed and characterized an intracellular phagocytosis-stimulating treatment in the CD47-SIRPαpathway.We loaded a novel monocyte/macrophage-selective nanoparticle carrier system with a small molecule enzymatic inhibitor that is released in a pH-dependent manner to stimulate macrophage efferocytosis of apoptotic cell debris via the CD47-SIRPαsignaling pathway.We demonstrated that single-walled carbon nanotubes(SWNTs)can selectively deliver tyrosine phosphatase inhibitor 1(TPI)intracellularly to macrophages,which potently stimulates efferocytosis,and chemically characterized the nanocarrier.Thus,SWNT-delivered TPI can stimulate macrophage efferocytosis,with the potential to reduce or prevent atherosclerotic disease.
