Background:Only a few bladder cancer patients benefit from anti-programmed cell death protein 1/programmed cell death ligand 1 immunotherapy.The cluster of differentiation 47(CD47)plays an important role in tumor immu...Background:Only a few bladder cancer patients benefit from anti-programmed cell death protein 1/programmed cell death ligand 1 immunotherapy.The cluster of differentiation 47(CD47)plays an important role in tumor immune evasion.CD47 is a highly glycosylated protein,however,the mechanisms governing CD47 glycosylation and its potential role in immunosuppression are unclear.Therefore,this study aimed to evaluate the function of CD47 glycosylation in bladder cancer.Methods:Western blotting,immunohistochemistry,and flow cytometry were used to measure protein expression,protein-protein interactions,and phagocytosis in bladder cancer.A murine model was employed to investigate the impact of mannosidase alpha class 1B member 1(MAN1B1)modification of CD47 on anti-phagocytosis in vivo.An ex vivo model,patient-derived tumor-like cell clusters,was used to examine the effect of targeting MAN1B1 on phagocytosis.Results:Our research identified that aberrant CD47 glycosylation was responsible for its immunosuppression.The glycosyltransferase MAN1B1 responsible for CD47 glycosylation was highly expressed in bladder cancer.Abnormal activation of extracellular signal-regulated kinase(ERK)was significantly associated with MAN1B1 stability by regulating the interaction between MAN1B1 and the E3 ubiquitin ligase HMG-CoA reductase degradation 1(HRD1).Mechanistically,abnormally activated ERK stabilized MAN1B1,resulting in the glycosylation of CD47 and facilitating immune evasion by enhancing its interaction with signal-regulatory protein alpha(SIRP-α).In vitro and in vivo experiments demonstrated that MAN1B1 knockout weakened CD47-mediated anti-phagocytosis.MAN1B1 inhibitors promoted phagocytosis without causing anemia,offering a safe alternative to anti-CD47 therapy.Conclusions:This comprehensive analysis uncovered that ERK activation stabilizes MAN1B1 by regulating the interaction between MAN1B1 and HRD1,facilitates immune evasion via CD47 glycosylation,and presents new potential targets and strategies for cancer immunotherapy that do not cause anemia.展开更多
Murray cod(Maccullochella peelii)is a freshwater percichthyid fish that has a high market value and potential for culture in recirculating aquaculture systems(RASs).Illumination is an important environmental factor th...Murray cod(Maccullochella peelii)is a freshwater percichthyid fish that has a high market value and potential for culture in recirculating aquaculture systems(RASs).Illumination is an important environmental factor that affects the growth and physiological condition of fishes.A comprehensive understanding of the relationship between the light factors and the growth,nutrient composition and stress response of juvenile Murray cod in RAS is important to achieve satisfactory theoretical and practical aquaculture performance.Juvenile Murray cod were randomly assigned to nine RAS tanks with a volume of 1.5 m^(3),with each tank containing 120 fishes(3.5±0.5g).The fish were cultured for 120 days under different light intensities(1200,2400,and 3600 lx)and photoperiods(12L:12D,18L:6D and 24L:0D).The results showed that the final weight and feed conversion ratio of Murray cod under a light intensity of 1200 lx were significantly better than those under 3600 lx(P<0.05).The serum total protein and globulin were higher at the light intensity of 1200lx,compared to other light intensities.The blood urea nitrogen level improved with increasing light intensity at LD12:12 and LD18:6 of photoperiod,as well as the superoxide dismutase level improved significantly from LD18:6 to LD24:0 of photoperiod.The findings indicated that the optimal light condition to enhance growth at juvenile stage is 1200 lx of the light intensity and LD18:6 of the photoperiod.展开更多
基金the Zhejiang Provincial Natural Science Foundation of China(grant Y24H160082 to Yanlan Yu)the National Natural Science Foundation of China(grant 32000799 to Jie Zhang,grant 81972367 to Yicheng Chen)+1 种基金Zhejiang Province key research and development program(grant 2021C03062 to Guoqing Ding,grant 2023C03010 to Yili Fu)the Zhejiang Provincial Natural Science Foundation of China(grant LQ21H160028 to Fengbin Gao,grant LY23H050004 to HaiyangWu).
文摘Background:Only a few bladder cancer patients benefit from anti-programmed cell death protein 1/programmed cell death ligand 1 immunotherapy.The cluster of differentiation 47(CD47)plays an important role in tumor immune evasion.CD47 is a highly glycosylated protein,however,the mechanisms governing CD47 glycosylation and its potential role in immunosuppression are unclear.Therefore,this study aimed to evaluate the function of CD47 glycosylation in bladder cancer.Methods:Western blotting,immunohistochemistry,and flow cytometry were used to measure protein expression,protein-protein interactions,and phagocytosis in bladder cancer.A murine model was employed to investigate the impact of mannosidase alpha class 1B member 1(MAN1B1)modification of CD47 on anti-phagocytosis in vivo.An ex vivo model,patient-derived tumor-like cell clusters,was used to examine the effect of targeting MAN1B1 on phagocytosis.Results:Our research identified that aberrant CD47 glycosylation was responsible for its immunosuppression.The glycosyltransferase MAN1B1 responsible for CD47 glycosylation was highly expressed in bladder cancer.Abnormal activation of extracellular signal-regulated kinase(ERK)was significantly associated with MAN1B1 stability by regulating the interaction between MAN1B1 and the E3 ubiquitin ligase HMG-CoA reductase degradation 1(HRD1).Mechanistically,abnormally activated ERK stabilized MAN1B1,resulting in the glycosylation of CD47 and facilitating immune evasion by enhancing its interaction with signal-regulatory protein alpha(SIRP-α).In vitro and in vivo experiments demonstrated that MAN1B1 knockout weakened CD47-mediated anti-phagocytosis.MAN1B1 inhibitors promoted phagocytosis without causing anemia,offering a safe alternative to anti-CD47 therapy.Conclusions:This comprehensive analysis uncovered that ERK activation stabilizes MAN1B1 by regulating the interaction between MAN1B1 and HRD1,facilitates immune evasion via CD47 glycosylation,and presents new potential targets and strategies for cancer immunotherapy that do not cause anemia.
基金study was provided by the National Key Research and Development Project:Precise Pond Culture Technology and Aquatic Product Quality Improvement Model"(2019YFD0900303)the Shanghai Agriculture Applied Technology Development Program,China(Grant No.X20210301)+1 种基金the China-ASEAN Maritime Cooperation Fund(China-ASEAN Center for Joint Research and Promotion of Marine Aquaculture Technology)China Postdoctoral Science Foundation Funded Project(Project No.:2018M641984)。
文摘Murray cod(Maccullochella peelii)is a freshwater percichthyid fish that has a high market value and potential for culture in recirculating aquaculture systems(RASs).Illumination is an important environmental factor that affects the growth and physiological condition of fishes.A comprehensive understanding of the relationship between the light factors and the growth,nutrient composition and stress response of juvenile Murray cod in RAS is important to achieve satisfactory theoretical and practical aquaculture performance.Juvenile Murray cod were randomly assigned to nine RAS tanks with a volume of 1.5 m^(3),with each tank containing 120 fishes(3.5±0.5g).The fish were cultured for 120 days under different light intensities(1200,2400,and 3600 lx)and photoperiods(12L:12D,18L:6D and 24L:0D).The results showed that the final weight and feed conversion ratio of Murray cod under a light intensity of 1200 lx were significantly better than those under 3600 lx(P<0.05).The serum total protein and globulin were higher at the light intensity of 1200lx,compared to other light intensities.The blood urea nitrogen level improved with increasing light intensity at LD12:12 and LD18:6 of photoperiod,as well as the superoxide dismutase level improved significantly from LD18:6 to LD24:0 of photoperiod.The findings indicated that the optimal light condition to enhance growth at juvenile stage is 1200 lx of the light intensity and LD18:6 of the photoperiod.
