Background: Cervical cancer is the second most common cancer in women worldwide [1]. Photodynamic therapy has been used for cervical intraepithelial neoplasia with good responses, but few studies have used newer photo...Background: Cervical cancer is the second most common cancer in women worldwide [1]. Photodynamic therapy has been used for cervical intraepithelial neoplasia with good responses, but few studies have used newer phototherapeutics. We evaluated the effectiveness of photodynamic therapy using Pc 4 in vitro and in vivo against human cervical cancer cells. Methods: CaSki and ME-180 cancer cells were grown as monolayers and spheroids. Cell growth and cytotoxicity were measured using a methylthiazol tetrazolium assay. Pc 4 cellular uptake and intracellular distribution were determined. For in vitro Pc 4 photodynamic therapy, cells were irradiated at 667 nm at a fluence of 2.5 J/cm<sup>2</sup> at 48 h. SCID mice were implanted with CaSki and ME-180 cells both subcutaneously and intracervically. Forty-eight hours after Pc 4 photodynamic therapy was administered at 75 and 150 J/cm<sup>2</sup>. Results: The IC<sub>50</sub>s for Pc 4 and Pc 4 photodynamic therapy for CaSki and ME-180 cells as monolayers were, 7.6 μM and 0.016 μM and >10 μM and 0.026 μM;as spheroids, IC<sub>50</sub>s of Pc 4 photodynamic therapy were, 0.26 μM and 0.01 μM. Pc 4 was taken up within cells and widely distributed in tumors and tissues. Intracervical photodynamic therapy resulted in tumor death, however mice died due to gastrointestinal toxicity. Photodynamic therapy resulted in subcutaneous tumor death and growth delay. Conclusions: Pc 4 photodynamic therapy caused death within cervical cancer cells and xenografts, supporting development of Pc 4 photodynamic therapy for treatment of cervical cancer. Support: P30-CA47904, CTSI BaCCoR Pilot Program.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
Chondroitin sulfate proteoglycan-4(CSPG4) is a surface component of two key cell types(oligodendrocyte progenitor cells(OPCs) and myeloid cells) present in lysolecithin-induced lesions in mouse spinal cord.Two t...Chondroitin sulfate proteoglycan-4(CSPG4) is a surface component of two key cell types(oligodendrocyte progenitor cells(OPCs) and myeloid cells) present in lysolecithin-induced lesions in mouse spinal cord.Two types of CSPG4 manipulations have been used to study the roles of these cells in myelin damage and repair:(1) OPC and myeloid-specific ablation of CSPG4,and(2) transplantation of enhanced green fluorescent protein(EGFP)-labeled progenitors to distinguish between bone marrow-derived macrophages and resident microglia.Ablation of CSPG4 in OPCs does not affect myelin damage,but decreases myelin repair,due to reduced proliferation of CSPG4-null OPCs that diminishes generation of mature oligodendrocytes for remyelination.Ablation of CSPG4 in myeloid cells greatly decreases recruitment of macrophages to spinal cord lesions,resulting in smaller initial lesions,but also in significantly diminished myelin repair.In the absence of macrophage recruitment,OPC proliferation is greatly impaired,again leading to decreased generation of myelinating oligodendrocytes.Macrophages may promote OPC proliferation via phagocytosis of myelin debris and/or secretion of factors that stimulate OPC mitosis.Microglia are not able to substitute for macrophages in promoting OPC proliferation.An additional feature of lesions in myeloid-specific CSPG4 null mice is the persistence of poorly-differentiated platelet-derived growth factor receptor α(PDGFRα) + macrophages that may prolong damage.展开更多
The food colorant Red 40 is an environmental risk factor for colitis development in mice with increased expression of interleukin(IL)-23.This immune response is mediated by CD4^(+)T cells,but mechanistic insights into...The food colorant Red 40 is an environmental risk factor for colitis development in mice with increased expression of interleukin(IL)-23.This immune response is mediated by CD4^(+)T cells,but mechanistic insights into how these CD4^(+)T cells trigger andperpetuate colitis have remained elusive.Here,using single-cell transcriptomic analysis,we found that several CD4^(+)T-cell subsetsare present in the intestines of colitic mice,including an interferon(IFN)-γ-producing subset.In vivo challenge of primed mice withRed 40 promoted rapid activation of CD4^(+)T cells and caused marked intestinal epithelial cell(IEC)apoptosis that was attenuated bydepletion of CD4^(+)cells and blockade of IFN-γ.Ex vivo experiments showed that intestinal CD4^(+)T cells from colitic mice directlypromoted apoptosis of IECs and intestinal enteroids.CD4^(+)T cell-mediated cytotoxicity was contact-dependent and required FasL,which promoted caspase-dependent cell death in target IECs.Genetic ablation of IFN-γconstrained IL-23-and Red 40-inducedcolitis development,and blockade of IFN-γinhibited epithelial cell death in vivo.These results advance the understanding of themechanisms regulating colitis development caused by IL-23 and food colorants and identify IFN-γ^(+)cytotoxic CD4^(+)T cells as a newpotential therapeutic target for colitis.展开更多
In a study recently published in Nature,Lawson et al.confirm several pathways previously implicated in tumor cell sensitivity to cytotoxic T lymphocytes(CTLs),and identify autophagy as an additional pathway involved i...In a study recently published in Nature,Lawson et al.confirm several pathways previously implicated in tumor cell sensitivity to cytotoxic T lymphocytes(CTLs),and identify autophagy as an additional pathway involved in mediating tumor immune evasion.1 Autophagy is a regulated and conserved multistep process that enables cells to degrade and recycle intracellular components and is critical for cellular homeostasis.Autophagy has recently been associated with tumor immune evasion in pancreatic ductal adenocarcinoma,2 whereby this process selectively targeted the degradation of major histocompatibility complex I(MHC-I),limiting recognition,and anti-tumor responses by CTLs.Here,Lawson et al.identify an additional role for autophagy in tumor immune evasion,by mediating resistance to cytokine-mediated cell death.展开更多
In a study recently published in Nature,Lawson et al.confirm several pathways previously implicated in tumor cell sensitivity to cytotoxic T lymphocytes(CTLs),and identify autophagy as an additional pathway involved i...In a study recently published in Nature,Lawson et al.confirm several pathways previously implicated in tumor cell sensitivity to cytotoxic T lymphocytes(CTLs),and identify autophagy as an additional pathway involved in mediating tumor immune evasion.1 Autophagy is a regulated and conserved multistep process that enables cells to degrade and recycle intracellular components and is critical for cellular homeostasis.Autophagy has recently been associated with tumor immune evasion in pancreatic ductal adenocarcinoma,2 whereby this process selectively targeted the degradation of major histocompatibility complex I(MHC-I),limiting recognition,and anti-tumor responses by CTLs.Here,Lawson et al.identify an additional role for autophagy in tumor immune evasion,by mediating resistance to cytokine-mediated cell death.展开更多
文摘Background: Cervical cancer is the second most common cancer in women worldwide [1]. Photodynamic therapy has been used for cervical intraepithelial neoplasia with good responses, but few studies have used newer phototherapeutics. We evaluated the effectiveness of photodynamic therapy using Pc 4 in vitro and in vivo against human cervical cancer cells. Methods: CaSki and ME-180 cancer cells were grown as monolayers and spheroids. Cell growth and cytotoxicity were measured using a methylthiazol tetrazolium assay. Pc 4 cellular uptake and intracellular distribution were determined. For in vitro Pc 4 photodynamic therapy, cells were irradiated at 667 nm at a fluence of 2.5 J/cm<sup>2</sup> at 48 h. SCID mice were implanted with CaSki and ME-180 cells both subcutaneously and intracervically. Forty-eight hours after Pc 4 photodynamic therapy was administered at 75 and 150 J/cm<sup>2</sup>. Results: The IC<sub>50</sub>s for Pc 4 and Pc 4 photodynamic therapy for CaSki and ME-180 cells as monolayers were, 7.6 μM and 0.016 μM and >10 μM and 0.026 μM;as spheroids, IC<sub>50</sub>s of Pc 4 photodynamic therapy were, 0.26 μM and 0.01 μM. Pc 4 was taken up within cells and widely distributed in tumors and tissues. Intracervical photodynamic therapy resulted in tumor death, however mice died due to gastrointestinal toxicity. Photodynamic therapy resulted in subcutaneous tumor death and growth delay. Conclusions: Pc 4 photodynamic therapy caused death within cervical cancer cells and xenografts, supporting development of Pc 4 photodynamic therapy for treatment of cervical cancer. Support: P30-CA47904, CTSI BaCCoR Pilot Program.
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 National Institutes of Health R01 CA095287(WBS)Sanford Burnham Prebys Medical Discovery Institute Lab Funding Initiative(WBS)
文摘Chondroitin sulfate proteoglycan-4(CSPG4) is a surface component of two key cell types(oligodendrocyte progenitor cells(OPCs) and myeloid cells) present in lysolecithin-induced lesions in mouse spinal cord.Two types of CSPG4 manipulations have been used to study the roles of these cells in myelin damage and repair:(1) OPC and myeloid-specific ablation of CSPG4,and(2) transplantation of enhanced green fluorescent protein(EGFP)-labeled progenitors to distinguish between bone marrow-derived macrophages and resident microglia.Ablation of CSPG4 in OPCs does not affect myelin damage,but decreases myelin repair,due to reduced proliferation of CSPG4-null OPCs that diminishes generation of mature oligodendrocytes for remyelination.Ablation of CSPG4 in myeloid cells greatly decreases recruitment of macrophages to spinal cord lesions,resulting in smaller initial lesions,but also in significantly diminished myelin repair.In the absence of macrophage recruitment,OPC proliferation is greatly impaired,again leading to decreased generation of myelinating oligodendrocytes.Macrophages may promote OPC proliferation via phagocytosis of myelin debris and/or secretion of factors that stimulate OPC mitosis.Microglia are not able to substitute for macrophages in promoting OPC proliferation.An additional feature of lesions in myeloid-specific CSPG4 null mice is the persistence of poorly-differentiated platelet-derived growth factor receptor α(PDGFRα) + macrophages that may prolong damage.
基金This work was supported by grants from the National Institutes of Health(R01 DK 110352 and DK 121009 to SAL)a Career Development Award(634253)from the Crohn’s&Colitis Foundation of America(CCFA)(to LC)The funders of the study had no involvement in the study design,data collection,data analysis,interpretation,writing of the report,or decision to submit the paper for publication。
文摘The food colorant Red 40 is an environmental risk factor for colitis development in mice with increased expression of interleukin(IL)-23.This immune response is mediated by CD4^(+)T cells,but mechanistic insights into how these CD4^(+)T cells trigger andperpetuate colitis have remained elusive.Here,using single-cell transcriptomic analysis,we found that several CD4^(+)T-cell subsetsare present in the intestines of colitic mice,including an interferon(IFN)-γ-producing subset.In vivo challenge of primed mice withRed 40 promoted rapid activation of CD4^(+)T cells and caused marked intestinal epithelial cell(IEC)apoptosis that was attenuated bydepletion of CD4^(+)cells and blockade of IFN-γ.Ex vivo experiments showed that intestinal CD4^(+)T cells from colitic mice directlypromoted apoptosis of IECs and intestinal enteroids.CD4^(+)T cell-mediated cytotoxicity was contact-dependent and required FasL,which promoted caspase-dependent cell death in target IECs.Genetic ablation of IFN-γconstrained IL-23-and Red 40-inducedcolitis development,and blockade of IFN-γinhibited epithelial cell death in vivo.These results advance the understanding of themechanisms regulating colitis development caused by IL-23 and food colorants and identify IFN-γ^(+)cytotoxic CD4^(+)T cells as a newpotential therapeutic target for colitis.
基金A.J.F.was supported by scholarships from the Australian Government,Steer North Australia,and the Peter MacCallum Cancer Foundation.E.J.L.and J.O.were supported by National Health and M。
文摘In a study recently published in Nature,Lawson et al.confirm several pathways previously implicated in tumor cell sensitivity to cytotoxic T lymphocytes(CTLs),and identify autophagy as an additional pathway involved in mediating tumor immune evasion.1 Autophagy is a regulated and conserved multistep process that enables cells to degrade and recycle intracellular components and is critical for cellular homeostasis.Autophagy has recently been associated with tumor immune evasion in pancreatic ductal adenocarcinoma,2 whereby this process selectively targeted the degradation of major histocompatibility complex I(MHC-I),limiting recognition,and anti-tumor responses by CTLs.Here,Lawson et al.identify an additional role for autophagy in tumor immune evasion,by mediating resistance to cytokine-mediated cell death.
基金A.J.F.was supported by scholarships from the Australian Government,Steer North Australiathe Peter MacCallum Cancer Foundation.E.J.L.and J.O.were supported by National Health and Medical Research Council and National Breast Cancer Foundation project grants.
文摘In a study recently published in Nature,Lawson et al.confirm several pathways previously implicated in tumor cell sensitivity to cytotoxic T lymphocytes(CTLs),and identify autophagy as an additional pathway involved in mediating tumor immune evasion.1 Autophagy is a regulated and conserved multistep process that enables cells to degrade and recycle intracellular components and is critical for cellular homeostasis.Autophagy has recently been associated with tumor immune evasion in pancreatic ductal adenocarcinoma,2 whereby this process selectively targeted the degradation of major histocompatibility complex I(MHC-I),limiting recognition,and anti-tumor responses by CTLs.Here,Lawson et al.identify an additional role for autophagy in tumor immune evasion,by mediating resistance to cytokine-mediated cell death.