Introduction:Butyrophilins(BTNs)belong to the immunoglobulin superfamily;they play crucial roles in immune regulation,especially inγδ T cell activation.While their expression has been studied in solid tumours,their ...Introduction:Butyrophilins(BTNs)belong to the immunoglobulin superfamily;they play crucial roles in immune regulation,especially inγδ T cell activation.While their expression has been studied in solid tumours,their involvement in hematologic malignancies remains poorly understood.Objectives:We hypothesised that BTNs are dysregulated in chronic lymphocytic leukaemia(CLL),contributing toγδT cell dysfunction and potentially influencing disease progression.Methods:In this study,we analyzed publicly available microarray and RNA-seq datasets to investigate the expression patterns of BTN genes in CLL.Results:Our findings reveal significant dysregulation of BTN gene expression in CLL,with BTN2A1,BTN3A1,BTN3A2,and BTN3A3 being markedly downregulated in peripheral blood mononuclear cells(PBMCs)and bone marrow samples from CLL patients compared to healthy volunteers,while BTN1A1 was upregulated.Furthermore,BTN2A2 was selectively downregulated in neoplastic B cells,whereas BTN3A1 was upregulated in T cells from CLL patients compared to healthy volunteers.Notably,lower BTN expression was associated with an unmutated IGVH status and male sex.Kaplan-Meier survival analysis demonstrated that higher expression of BTN2A1,BTN3A1,BTN3A2,and BTN3A3 correlated with a significantly longer overall survival.Conclusions:Given the established role of BTN2A1 and BTN3A1 in the phosphoantigen-mediated activation of Vδ2γδ T cells,their downregulation may contribute toγδ T cell dysfunction in CLL.These results highlight the potential prognostic value of BTN gene expression in CLL and underscore the need for further studies exploring its mechanistic role in disease progression and immune evasion.展开更多
Vγ9Vδ2 T cells are specialized effector cells that have gained prominence as immunotherapy agents due to their ability to target and kill cells with altered pyrophosphate metabolites.In our effort to understand how ...Vγ9Vδ2 T cells are specialized effector cells that have gained prominence as immunotherapy agents due to their ability to target and kill cells with altered pyrophosphate metabolites.In our effort to understand how cancer cells evade the cell-killing activity of Vγ9Vδ2 T cells,we performed a comprehensive genome-scale CRISPR screening of cancer cells.We found that four molecules belonging to the butyrophilin(BTN)family,specifically BTN2A1,BTN3A1,BTN3A2,and BTN3A3,are critically important and play unique,nonoverlapping roles in facilitating the destruction of cancer cells by primary Vγ9Vδ2 T cells.The coordinated function of these BTN molecules was driven by synchronized gene expression,which was regulated by IFN-γsignaling and the RFX complex.Additionally,an enzyme called QPCTL was shown to play a key role in modifying the N-terminal glutamine of these BTN proteins and was found to be a crucial factor in Vγ9Vδ2 T cell killing of cancer cells.Through our research,we offer a detailed overview of the functional genomic mechanisms that underlie how cancer cells escape Vγ9Vδ2 T cells.Moreover,our findings shed light on the importance of the harmonized expression and function of gene family members in modulating T-cell activity.展开更多
BACKGROUND Pancreatic cancer(PC)is an aggressive malignancy.As a member of the BTN/BTNL family,BTNL9 has been identified as a tumor suppressor in breast cancer,lung adenocarcinoma,and colon cancer;however,its role and...BACKGROUND Pancreatic cancer(PC)is an aggressive malignancy.As a member of the BTN/BTNL family,BTNL9 has been identified as a tumor suppressor in breast cancer,lung adenocarcinoma,and colon cancer;however,its role and underlying mechanisms in PC remain to be elucidated.AIM To investigate the role of BTNL9 in the pathogenesis and development of PC.METHODS The difference of BTNL9 expression in cancer and adjacent normal tissues was analyzed by RNA sequencing data from a public database and tissue microarray detection.The relationship between BTNL9 expression and the prognosis of patients was also studied.The effects of BTNL9 on proliferation,metastasis,and cell cycle of PC cells were investigated by phenotypic experiments.The mechanism was investigated by RNA sequencing,western blotting,and immunofluorescence detection.RESULTS The mRNA and protein levels of BTNL9 in PC tissues were downregulated compared with normal tissues.Based on survival data from The Cancer Genome Atlas and tissue microarray,BTNL9 was an independent influencing factor for overall survival,and its low expression predicted a shortened overall survival of patients.In vitro,BTNL9 could inhibit cell proliferation and metastasis in both PANC-1 and MIA PaCa-2 cells and induce cell cycle arrest in G2/M phases.Downregulation of BTNL9 could activate the cell cycle signaling pathway.Furthermore,overexpression of BTNL9 could significantly inhibit the expression of cell division cycle 20(CDC20).Rescue experiments demonstrated that overexpression of CDC20 reversed the effect of BTNL9 on the proliferation,metastasis,and cell cycle of PC cells.CONCLUSION The expression of BTNL9 was downregulated in PC,and it has the prediction ability for prognosis.Functionally,BTNL9 exerted an anti-cancer effect by suppressing downstream CDC20 expression in PC.展开更多
The high cytotoxic activity of Vγ9Vδ2 T lymphocytes against tumor cells makes them useful candidates in anticancer therapies.However,the molecular mechanism of their activation by phosphoantigens(PAgs)is not complet...The high cytotoxic activity of Vγ9Vδ2 T lymphocytes against tumor cells makes them useful candidates in anticancer therapies.However,the molecular mechanism of their activation by phosphoantigens(PAgs)is not completely known.Many studies have depicted the mechanism of Vγ9Vδ2 T-cell activation by PAg-sensed accessory cells,such as immune presenting cells or tumor cells.In this study,we demonstrated that pure resting Vγ9Vδ2 T lymphocytes can self-activate through exogenous PAgs,involving their TCR and the butyrophilins BTN3A1 and BTN2A1.This is the first time that these three molecules,concurrently expressed at the plasma membrane of Vγ9Vδ2 T cells,have been shown to be involved together on the same and unique T cell during PAg activation.Moreover,the use of probucol to stimulate the inhibition of this self-activation prompted us to propose that ABCA-1 could be implicated in the transfer of exogenous PAgs inside Vγ9Vδ2 T cells before activating them through membrane clusters formed byγ9TCR,BTN3A1 and BTN2A1.The self-activation of Vγ9Vδ2 T cells,which leads to self-killing,can therefore participate in the failure ofγδT cell-based therapies with exogenous PAgs and should be taken into account.展开更多
基金funded by the National Science Center in Poland,Preludium grant number 2019/35/N/NZ6/02973.
文摘Introduction:Butyrophilins(BTNs)belong to the immunoglobulin superfamily;they play crucial roles in immune regulation,especially inγδ T cell activation.While their expression has been studied in solid tumours,their involvement in hematologic malignancies remains poorly understood.Objectives:We hypothesised that BTNs are dysregulated in chronic lymphocytic leukaemia(CLL),contributing toγδT cell dysfunction and potentially influencing disease progression.Methods:In this study,we analyzed publicly available microarray and RNA-seq datasets to investigate the expression patterns of BTN genes in CLL.Results:Our findings reveal significant dysregulation of BTN gene expression in CLL,with BTN2A1,BTN3A1,BTN3A2,and BTN3A3 being markedly downregulated in peripheral blood mononuclear cells(PBMCs)and bone marrow samples from CLL patients compared to healthy volunteers,while BTN1A1 was upregulated.Furthermore,BTN2A2 was selectively downregulated in neoplastic B cells,whereas BTN3A1 was upregulated in T cells from CLL patients compared to healthy volunteers.Notably,lower BTN expression was associated with an unmutated IGVH status and male sex.Kaplan-Meier survival analysis demonstrated that higher expression of BTN2A1,BTN3A1,BTN3A2,and BTN3A3 correlated with a significantly longer overall survival.Conclusions:Given the established role of BTN2A1 and BTN3A1 in the phosphoantigen-mediated activation of Vδ2γδ T cells,their downregulation may contribute toγδ T cell dysfunction in CLL.These results highlight the potential prognostic value of BTN gene expression in CLL and underscore the need for further studies exploring its mechanistic role in disease progression and immune evasion.
基金funding from the National Science Foundation of China(31930016)the Peking-Tsinghua Center for Life Sciences+4 种基金ZW received funding from the State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases(2024KF00001)the National Science Foundation of China(82350119)CCW received funding from the Talent Introduction Funds from the Chinese Academy of Medical Science(2022-RC310-10)the National Science Foundation of China(32150005)the Research Funds from Health@InnoHK Program,launched by the Innovation Technology Commission of the Hong Kong Special Administrative Region.
文摘Vγ9Vδ2 T cells are specialized effector cells that have gained prominence as immunotherapy agents due to their ability to target and kill cells with altered pyrophosphate metabolites.In our effort to understand how cancer cells evade the cell-killing activity of Vγ9Vδ2 T cells,we performed a comprehensive genome-scale CRISPR screening of cancer cells.We found that four molecules belonging to the butyrophilin(BTN)family,specifically BTN2A1,BTN3A1,BTN3A2,and BTN3A3,are critically important and play unique,nonoverlapping roles in facilitating the destruction of cancer cells by primary Vγ9Vδ2 T cells.The coordinated function of these BTN molecules was driven by synchronized gene expression,which was regulated by IFN-γsignaling and the RFX complex.Additionally,an enzyme called QPCTL was shown to play a key role in modifying the N-terminal glutamine of these BTN proteins and was found to be a crucial factor in Vγ9Vδ2 T cell killing of cancer cells.Through our research,we offer a detailed overview of the functional genomic mechanisms that underlie how cancer cells escape Vγ9Vδ2 T cells.Moreover,our findings shed light on the importance of the harmonized expression and function of gene family members in modulating T-cell activity.
基金Supported by Sichuan Medical Association Research Project,No.Q2024049Clinical Science Research Fund of Chengdu Medical College,No.24 LHLNYX1-03+1 种基金Chengdu Medical College Elite Peak Program,No.2024qnGzn04Key Clinical Specialty of Sichuan Province,No.YS00109.
文摘BACKGROUND Pancreatic cancer(PC)is an aggressive malignancy.As a member of the BTN/BTNL family,BTNL9 has been identified as a tumor suppressor in breast cancer,lung adenocarcinoma,and colon cancer;however,its role and underlying mechanisms in PC remain to be elucidated.AIM To investigate the role of BTNL9 in the pathogenesis and development of PC.METHODS The difference of BTNL9 expression in cancer and adjacent normal tissues was analyzed by RNA sequencing data from a public database and tissue microarray detection.The relationship between BTNL9 expression and the prognosis of patients was also studied.The effects of BTNL9 on proliferation,metastasis,and cell cycle of PC cells were investigated by phenotypic experiments.The mechanism was investigated by RNA sequencing,western blotting,and immunofluorescence detection.RESULTS The mRNA and protein levels of BTNL9 in PC tissues were downregulated compared with normal tissues.Based on survival data from The Cancer Genome Atlas and tissue microarray,BTNL9 was an independent influencing factor for overall survival,and its low expression predicted a shortened overall survival of patients.In vitro,BTNL9 could inhibit cell proliferation and metastasis in both PANC-1 and MIA PaCa-2 cells and induce cell cycle arrest in G2/M phases.Downregulation of BTNL9 could activate the cell cycle signaling pathway.Furthermore,overexpression of BTNL9 could significantly inhibit the expression of cell division cycle 20(CDC20).Rescue experiments demonstrated that overexpression of CDC20 reversed the effect of BTNL9 on the proliferation,metastasis,and cell cycle of PC cells.CONCLUSION The expression of BTNL9 was downregulated in PC,and it has the prediction ability for prognosis.Functionally,BTNL9 exerted an anti-cancer effect by suppressing downstream CDC20 expression in PC.
基金This work was funded by INSERM,CNRS,the University Hospital of Bordeaux,and Toulouse III University.We acknowledge ImCheck for providing the 103.2 antibody and the 7.48 antibody.We are grateful to our healthcare professionals for their boundless efforts during the COVID-19 crisis.
文摘The high cytotoxic activity of Vγ9Vδ2 T lymphocytes against tumor cells makes them useful candidates in anticancer therapies.However,the molecular mechanism of their activation by phosphoantigens(PAgs)is not completely known.Many studies have depicted the mechanism of Vγ9Vδ2 T-cell activation by PAg-sensed accessory cells,such as immune presenting cells or tumor cells.In this study,we demonstrated that pure resting Vγ9Vδ2 T lymphocytes can self-activate through exogenous PAgs,involving their TCR and the butyrophilins BTN3A1 and BTN2A1.This is the first time that these three molecules,concurrently expressed at the plasma membrane of Vγ9Vδ2 T cells,have been shown to be involved together on the same and unique T cell during PAg activation.Moreover,the use of probucol to stimulate the inhibition of this self-activation prompted us to propose that ABCA-1 could be implicated in the transfer of exogenous PAgs inside Vγ9Vδ2 T cells before activating them through membrane clusters formed byγ9TCR,BTN3A1 and BTN2A1.The self-activation of Vγ9Vδ2 T cells,which leads to self-killing,can therefore participate in the failure ofγδT cell-based therapies with exogenous PAgs and should be taken into account.
