Tyrosine phosphatase SHP2 is a promising drug target in cancer immunotherapy due to its bidirectional role in both tumor growth promotion and T-cell inactivation. Its allosteric inhibitor SHP099 is known to inhibit ca...Tyrosine phosphatase SHP2 is a promising drug target in cancer immunotherapy due to its bidirectional role in both tumor growth promotion and T-cell inactivation. Its allosteric inhibitor SHP099 is known to inhibit cancer cell growth both in vitro and in vivo. However, whether SHP099-mediated SHP2 inhibition retards tumor growth in vivo via anti-tumor immunity remains elusive. To address this, a CT-26 colon cancer xenograft model was established in mice since this cell line is insensitive to SHP099.Consequently, SHP099 minimally affected CT-26 tumor growth in immuno-deficient nude mice, but significantly decreased the tumor burden in CT-26 tumor-bearing mice with intact immune system.SHP099 augmented anti-tumor immunity, as shown by the elevated proportion of CD8tIFN-γtT cells and the upregulation of cytotoxic T-cell related genes including Granzyme B andPerforin, which decreased the tumor load. In addition, tumor growth in mice with SHP2-deficient T-cells was markedly slowed down because of enhanced anti-tumor responses. Finally, the combination of SHP099 and antiPD-1 antibody showed a higher therapeutic efficacy than either monotherapy in controlling tumor growthin two colon cancer xenograft models, indicating that these agents complement each other. Our study suggests that SHP2 inhibitor SHP099 is a promising candidate drug for cancer immunotherapy.展开更多
Chronic infection with Helicobacter pylori cagA-positive strains is the strongest risk factor for gastric cancer.The cagA gene product,CagA,is delivered into gastric epithelial cells via the bacterial type IV secretio...Chronic infection with Helicobacter pylori cagA-positive strains is the strongest risk factor for gastric cancer.The cagA gene product,CagA,is delivered into gastric epithelial cells via the bacterial type IV secretion system.Delivered CagA then undergoes tyrosine phosphorylation at the Glu-Pro-Ile-Tyr-Ala(EPIYA)motifs in its C-terminal region and acts as an oncogenic scaffold protein that physically interacts with multiple host signaling proteins in both tyrosine phosphorylation-dependent and-independent manners.Analysis of CagA using in vitro cultured gastric epithelial cells has indicated that the nonphysiological scaffolding actions of CagA cell-autonomously promote the malignant transformation of the cells by endowing the cells with multiple phenotypic cancer hallmarks:sustained proliferation,evasion of growth suppressors,invasiveness,resistance to cell death,and genomic instability.Transgenic expression of CagA in mice leads to in vivo oncogenic action of CagA without any overt inflammation.The in vivo oncogenic activity of CagA is further potentiated in the presence of chronic inflammation.Since Helicobacter pylori infection triggers a proinflammatory response in host cells,a feedforward stimulation loop that augments the oncogenic actions of CagA and inflammation is created in CagA-injected gastric mucosa.Given that Helicobacter pylori is no longer colonized in established gastric cancer lesions,the multistep nature of gastric cancer development should include a“hit-and-run”process of CagA action.Thus,acquisition of genetic and epigenetic alterations that compensate for CagA-directed cancer hallmarks may be required for completion of the“hit-and-run”process of gastric carcinogenesis.展开更多
Cellular biological activities are tightly controlled by intracellular signaling processes initiated by extracellular signals. Protein tyrosine phosphatases, which remove phosphate groups from phosphorylated signaling...Cellular biological activities are tightly controlled by intracellular signaling processes initiated by extracellular signals. Protein tyrosine phosphatases, which remove phosphate groups from phosphorylated signaling molecules, play equally important tyrosine roles as protein tyrosine kinases in signal transduction. SHP-2, a cytoplajsmic SH2 domain containing protein tyrosine phosphatase, is involved in the signaling pathways of a variety of growth factors and cytokines. Recent studies have clearly demonstrated that this phosphatase plays an important role in transducing signal relay from the cell surface to the nucleus, and is a critical intracellular regulator in mediating cell proliferation and differentiation.展开更多
Objective:The tyrosine phosphatase SHP2 has a dual role in cancer initiation and progression in a tissue type-dependent manner.Several studies have linked SHP2 to the aggressive behavior of breast cancer cells and poo...Objective:The tyrosine phosphatase SHP2 has a dual role in cancer initiation and progression in a tissue type-dependent manner.Several studies have linked SHP2 to the aggressive behavior of breast cancer cells and poorer outcomes in people with cancer.Nevertheless,the mechanistic details of how SHP2 promotes breast cancer progression remain largely undefined.Methods:The relationship between SHP2 expression and the prognosis of patients with breast cancer was investigated by using the TCGA and GEO databases.The expression of SHP2 in breast cancer tissues was analyzed by immunohistochemistry.CRISPR/Cas9 technology was used to generate SHP2-knockout breast cancer cells.Cell-counting kit-8,colony formation,cell cycle,and EdU incorporation assays,as well as a tumor xenograft model were used to examine the function of SHP2 in breast cancer proliferation.Quantitative RT-PCR,western blotting,immunofluorescence staining,and ubiquitination assays were used to explore the molecular mechanism through which SHP2 regulates breast cancer proliferation.Results:High SHP2 expression is correlated with poor prognosis in patients with breast cancer.SHP2 is required for the proliferation of breast cancer cellsin vitro and tumor growthin vivo through regulation of Cyclin D1 abundance,thereby accelerating cell cycle progression.Notably,SHP2 modulates the ubiquitin–proteasome-dependent degradation of Cyclin D1viathe PI3K/AKT/GSK3βsignaling pathway.SHP2 knockout attenuates the activation of PI3K/AKT signaling and causes the dephosphorylation and resultant activation of GSK3β.GSK3βthen mediates phosphorylation of Cyclin D1 at threonine 286,thereby promoting the translocation of Cyclin D1 from the nucleus to the cytoplasm and facilitating Cyclin D1 degradation through the ubiquitin–proteasome system.Conclusions:Our study uncovered the mechanism through which SHP2 regulates breast cancer proliferation.SHP2 may therefore potentially serve as a therapeutic target for breast cancer.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
基金supported by National Natural Science Foundation of China(Nos.81673436,21472091,81872877,81673437)Mountain-Climbing Talents Project of Nanjing University,the Open Fund of State Key Laboratory of Pharmaceutical Biotechnology,Nanjing University(No.KF-GN-201703,China)+1 种基金Open Project Program of Jiangsu Key Laboratory for Pharmacology and Safety Evaluation of Chinese Materia Medica(No.JKLPSE201802,China)the Project of the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD,China)
文摘Tyrosine phosphatase SHP2 is a promising drug target in cancer immunotherapy due to its bidirectional role in both tumor growth promotion and T-cell inactivation. Its allosteric inhibitor SHP099 is known to inhibit cancer cell growth both in vitro and in vivo. However, whether SHP099-mediated SHP2 inhibition retards tumor growth in vivo via anti-tumor immunity remains elusive. To address this, a CT-26 colon cancer xenograft model was established in mice since this cell line is insensitive to SHP099.Consequently, SHP099 minimally affected CT-26 tumor growth in immuno-deficient nude mice, but significantly decreased the tumor burden in CT-26 tumor-bearing mice with intact immune system.SHP099 augmented anti-tumor immunity, as shown by the elevated proportion of CD8tIFN-γtT cells and the upregulation of cytotoxic T-cell related genes including Granzyme B andPerforin, which decreased the tumor load. In addition, tumor growth in mice with SHP2-deficient T-cells was markedly slowed down because of enhanced anti-tumor responses. Finally, the combination of SHP099 and antiPD-1 antibody showed a higher therapeutic efficacy than either monotherapy in controlling tumor growthin two colon cancer xenograft models, indicating that these agents complement each other. Our study suggests that SHP2 inhibitor SHP099 is a promising candidate drug for cancer immunotherapy.
基金Our studies presented in this review article were supported by Grants-in-Aid for Scientific Research“S”(#16H06373 to M.H.),“A”(#22240085 and#25250016 to M.H.)and“C”(#19K05945 to A.T.-K.)Grants-in-Aid for Innovative Areas(#3205,#22114001,and#22114002 to M.H.)+4 种基金Grants-in-Aid for Japan Society for the Promotion of Science(JSPS)Fellows(#07J03878 to A.T.-K.and#19J12668 to C.T.K.)Grants-in-Aid for Young Scientists“B”(#24700965 and#15K18399 to A.T.-K.)the Graduate Program for Leaders in Life Innovation(GPLLI,to C.T.K.)from the Ministry of Education,Culture,Sports,Science and Technology(MEXT)of Japan,by CREST(#120200000396 to M.H.)Japan Science and Technology Agency,by the Uehara Memorial Foundation(#137 in 2016 to A.T.-K.)by the Max-Planck Society,Germany。
文摘Chronic infection with Helicobacter pylori cagA-positive strains is the strongest risk factor for gastric cancer.The cagA gene product,CagA,is delivered into gastric epithelial cells via the bacterial type IV secretion system.Delivered CagA then undergoes tyrosine phosphorylation at the Glu-Pro-Ile-Tyr-Ala(EPIYA)motifs in its C-terminal region and acts as an oncogenic scaffold protein that physically interacts with multiple host signaling proteins in both tyrosine phosphorylation-dependent and-independent manners.Analysis of CagA using in vitro cultured gastric epithelial cells has indicated that the nonphysiological scaffolding actions of CagA cell-autonomously promote the malignant transformation of the cells by endowing the cells with multiple phenotypic cancer hallmarks:sustained proliferation,evasion of growth suppressors,invasiveness,resistance to cell death,and genomic instability.Transgenic expression of CagA in mice leads to in vivo oncogenic action of CagA without any overt inflammation.The in vivo oncogenic activity of CagA is further potentiated in the presence of chronic inflammation.Since Helicobacter pylori infection triggers a proinflammatory response in host cells,a feedforward stimulation loop that augments the oncogenic actions of CagA and inflammation is created in CagA-injected gastric mucosa.Given that Helicobacter pylori is no longer colonized in established gastric cancer lesions,the multistep nature of gastric cancer development should include a“hit-and-run”process of CagA action.Thus,acquisition of genetic and epigenetic alterations that compensate for CagA-directed cancer hallmarks may be required for completion of the“hit-and-run”process of gastric carcinogenesis.
文摘Cellular biological activities are tightly controlled by intracellular signaling processes initiated by extracellular signals. Protein tyrosine phosphatases, which remove phosphate groups from phosphorylated signaling molecules, play equally important tyrosine roles as protein tyrosine kinases in signal transduction. SHP-2, a cytoplajsmic SH2 domain containing protein tyrosine phosphatase, is involved in the signaling pathways of a variety of growth factors and cytokines. Recent studies have clearly demonstrated that this phosphatase plays an important role in transducing signal relay from the cell surface to the nucleus, and is a critical intracellular regulator in mediating cell proliferation and differentiation.
基金This work was supported by grants from the National Natural S&ence Foundation of China(grant Nos.81372844,81472474,81772804 and 81903092)Tianjin Municipal Science and Technology Commission(grant No.16JCYBJC25400)+1 种基金Changjiang Researchers and Innovative Research Team(grant No.IRT_14R40)Postgraduate Innovation Fund of"13th Five-Year Comprehensive Investment,"Tianjin Medical University(grant No.YJSCX201716).
文摘Objective:The tyrosine phosphatase SHP2 has a dual role in cancer initiation and progression in a tissue type-dependent manner.Several studies have linked SHP2 to the aggressive behavior of breast cancer cells and poorer outcomes in people with cancer.Nevertheless,the mechanistic details of how SHP2 promotes breast cancer progression remain largely undefined.Methods:The relationship between SHP2 expression and the prognosis of patients with breast cancer was investigated by using the TCGA and GEO databases.The expression of SHP2 in breast cancer tissues was analyzed by immunohistochemistry.CRISPR/Cas9 technology was used to generate SHP2-knockout breast cancer cells.Cell-counting kit-8,colony formation,cell cycle,and EdU incorporation assays,as well as a tumor xenograft model were used to examine the function of SHP2 in breast cancer proliferation.Quantitative RT-PCR,western blotting,immunofluorescence staining,and ubiquitination assays were used to explore the molecular mechanism through which SHP2 regulates breast cancer proliferation.Results:High SHP2 expression is correlated with poor prognosis in patients with breast cancer.SHP2 is required for the proliferation of breast cancer cellsin vitro and tumor growthin vivo through regulation of Cyclin D1 abundance,thereby accelerating cell cycle progression.Notably,SHP2 modulates the ubiquitin–proteasome-dependent degradation of Cyclin D1viathe PI3K/AKT/GSK3βsignaling pathway.SHP2 knockout attenuates the activation of PI3K/AKT signaling and causes the dephosphorylation and resultant activation of GSK3β.GSK3βthen mediates phosphorylation of Cyclin D1 at threonine 286,thereby promoting the translocation of Cyclin D1 from the nucleus to the cytoplasm and facilitating Cyclin D1 degradation through the ubiquitin–proteasome system.Conclusions:Our study uncovered the mechanism through which SHP2 regulates breast cancer proliferation.SHP2 may therefore potentially serve as a therapeutic target for breast cancer.
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。
