A recently published study(Xin et al.,Prog Biochem Biophys,2026,53(2):431-441.DOI:10.3724/j.pibb.2025.0508)addresses the therapeutic challenges of pancreatic ductal adenocarcinoma(PDAC)by innovatively developing an or...A recently published study(Xin et al.,Prog Biochem Biophys,2026,53(2):431-441.DOI:10.3724/j.pibb.2025.0508)addresses the therapeutic challenges of pancreatic ductal adenocarcinoma(PDAC)by innovatively developing an orally administered nanogene delivery system.Designed to achieve in situ,efficient delivery of chimeric antigen receptor(CAR)genes to tumor sites,this approach offers a novel strategy for CAR-macrophage(CAR-M)based immunotherapy.Its key highlights are as follows.展开更多
Background:Glioblastoma(GBM)prognosis has seen little improvement over the past two decades.While immunotherapy has revolutionized cancer treatment,its impact on GBM remains limited.To characterize the evolving resear...Background:Glioblastoma(GBM)prognosis has seen little improvement over the past two decades.While immunotherapy has revolutionized cancer treatment,its impact on GBM remains limited.To characterize the evolving research landscape and identify future directions in GBM immunotherapy,we conducted a comprehensive bibliometric review.Methods:All literature related to immunotherapy in GBM from 1999 to 2024 was collected from the Web of Science Core Collection.CtieSpace and VOSviewer were used to conduct bibliometric analysis and visualize the data.Results:Bibliometric analysis identified 5038 publications authored by 23,335 researchers from 4699 institutions across 96 countries/regions,published in 945 journals.The United States produced the highest number of publications,while Switzerland achieved the highest average citation rate.Duke University led in institutional output and citations.John H Sampson was the most productive author,and Roger Stupp was the most cited.Frontiers in Immunology published the most papers,while Clinical Cancer Research was the most cited journal.Research focus centered on adoptive T cell therapy,particularly chimeric antigen receptor(CAR)-T cells with 572 dedicated publications.Within CAR-T research for GBM,the University of Pennsylvania was the leading institution,Frontiers in Immunology the predominant journal,and Christine E Brown(City of Hope National Medical Center)was the most prolific and cited author.Conclusions:There has been a growing interest in GBM immunotherapy over past decades.The United States is the dominant contributor.CAR-T therapy represents the primary research focus.Emerging strategies like chimeric antigen receptor-modified natural killer(CAR-NK)cells,chimeric antigen receptor-engineered macrophages(CAR-M),and cytomegalovirus-specific T cell receptor(CMV-TCR)T cells are gaining prominence,aiming to address limitations in antigen recognition inherent to CAR-T therapy for GBM.展开更多
Classical Hodgkin lymphoma(cHL)is characterized by rare Hodgkin/Reed-Sternberg(HRS)tumor cells that uniformly express cluster of differentiation(CD)30 molecules and orchestrate an immunosuppressive tumor microenvironm...Classical Hodgkin lymphoma(cHL)is characterized by rare Hodgkin/Reed-Sternberg(HRS)tumor cells that uniformly express cluster of differentiation(CD)30 molecules and orchestrate an immunosuppressive tumor microenvironment,making CD30 an attractive and selective therapeutic target.We summarize the biological rationale for CD30 as a therapeutic target and the preclinical and clinical evidence across major platforms:antibody-drug conjugates(brentuximab vedotin),monoclonal antibodies(including acimtamig and its combinations with Natural Killer cells),second-and third-generation chimeric antigen receptor(CAR)-T cells,and alternative modalities.Particular attention is given to standardized response assessment(IWG,Lugano,RECIL criteria),which enables appropriate cross-trial comparisons.Taken together,the data indicate that beyond the established role of brentuximab vedotin,CD30-directed CAR-T cells and bispecific antibodies demonstrate high activity in refractory cHL,especially when used with fludarabine-containing lymphodepletion,combined with programmed cell death 1(PD-1)receptor blockade as a strategy to eradicate minimal residual disease.Key challenges include durable effector-cell persistence and optimization of sequencing and combinations;notably,loss of CD30 as an escape mechanism appears uncommon.Integrating mechanistic insights into HRS biology with clinical trial data highlights strategies to enhance the efficacy,safety,and accessibility of CD30-directed immunotherapy.This review aims to provide a concise overview of CD30-targeted approaches in cHL,emphasizing therapeutic outcomes and the evolution of CAR-T technologies.展开更多
In the human spinal cord,astrocytes are the major glial cells.In vitro studies of human astrocytes are relatively simple.However,the straightforward nature of the in vitro environment and complex nature of the in vivo...In the human spinal cord,astrocytes are the major glial cells.In vitro studies of human astrocytes are relatively simple.However,the straightforward nature of the in vitro environment and complex nature of the in vivo environment limit comprehensive investigations into the structure and function of human astrocytes.Additionally,in vivo studies of human astrocytes are further limited by ethical issues.This means there is an urgent need to develop effective in vivo models to study the structure and function of human astrocytes.Here,we first directed human embryonic stem cells to differentiate into human spinal cord dorsal neural stem/progenitor cells in vitro,before transplanting these cells into the gray matter of the cervical spinal cord(C5-T2 segments)of naïve nude rats to create a chimeric human astrocytic rat spinal cord model.The transplanted human spinal cord dorsal neural stem/progenitor cells survived for at least 20 months in the spinal cord environment of the rats,with over 90%differentiating into human astrocytes.These human astrocytes were able to migrate caudally for long distances along the white matter towards the spinal cord.They expressed astrocytic cytoskeletal proteins and functionally-related proteins,suggesting their maturation and structural integration into the rat spinal cord.Thus,this humanized astrocyte chimeric rat spinal cord model provides a valuable tool for studying the role of human spinal cord astrocytes in various spinal diseases.展开更多
The paradigm of cancer treatment has been reshaped by chimeric antigen receptor(CAR)αβT cell therapy,yet its full potential remains constrained by fundamental limitations.While conventional CARαβT cells have achie...The paradigm of cancer treatment has been reshaped by chimeric antigen receptor(CAR)αβT cell therapy,yet its full potential remains constrained by fundamental limitations.While conventional CARαβT cells have achieved notable success in hematological malignancies,their broader application is hindered by the high cost and delays of autologous manufacturing,as well as the critical risk of graft-vs-host disease(GvHD).In addition,their efficacy against solid tumors is often compromised by the immunosuppressive tumor microenvironment(TME).As a promising solution,γδT cells are being developed as an alternative CAR platform.Their intrinsic ability to recognize transformed cells in a major histocompatibility complex(MHC)-independent manner minimizes the risk of GvHD and supports the creation of safe,effective allogeneic therapies.Building on this unique biology,the therapeutic efficacy of CARγδT cells is being enhanced through advanced engineering strategies.Key innovations include“armoring”technologies,such as cytokine secretion,checkpoint blockade,and metabolic rewiring,to overcome local immunosuppression and improve persistence,as well as the use of induced pluripotent stem cells(iPSCs)to generate standardized products from a renewable and consistent source.This expanding technological toolbox is also enabling novel applications beyond oncology.For example,chimeric autoantibody receptor(CAAR)constructs built onγδT cells integrate both classical and emerging insights into CARγδT cell therapy,highlighting innovations that are driving the field toward safer,more versatile,and longer-lasting treatments for cancer and autoimmunity.In light of these advancements,this review provides an overview of the current understanding ofγδT cell biology and highlights emerging engineering strategies that enhance the efficacy and durability of CARγδT cells across oncologic and autoimmune contexts.展开更多
Background:In patients with autoimmune hemolytic anemia(AIHA),the risk of relapse is high owing to persistent autoreactive B-cell activity.Multirefractory AIHA is a more advanced stage of disease that is defined by a ...Background:In patients with autoimmune hemolytic anemia(AIHA),the risk of relapse is high owing to persistent autoreactive B-cell activity.Multirefractory AIHA is a more advanced stage of disease that is defined by a lack of response to at least three lines of therapy.CD19-directed chimeric antigen receptor(CAR)T-cell therapy results in profound B-cell depletion and may be a useful approach to achieving drug-free remission in multirefractory AIHA.展开更多
Chimeric antigen receptor T(CAR-T)cell therapy represents a major advance in cancer immunotherapy,offering targeted treatment options,particularly for hematologic malignancies.This review comprehensively explores the ...Chimeric antigen receptor T(CAR-T)cell therapy represents a major advance in cancer immunotherapy,offering targeted treatment options,particularly for hematologic malignancies.This review comprehensively explores the structural evolution,production processes,and cytotoxic mechanisms underlying CAR-T function.Therapy involves engineering autologous T cells with synthetic receptors that allow major histocompatibility complex-independent recognition of tumor-associated antigens.Key structural components such as antigen recognition domains,spacers,transmembrane,and intracellular domains are optimized to enhance specificity,persistence,and cytotoxicity.CAR-T therapy exerts antitumor effects via granzyme-perforin degranulation,Fas/Fas ligand signaling,and cytokine secretion.Over time,the development of second-to fifth-generation CARs has incorporated costimulatory molecules,transcriptional regulation,and logic-gated control to improve efficacy and safety.Additionally,novel engineering strategies such as dual CARs,tandem CARs,SynNotch systems,and universal or inhibitory CARs have expanded antigen targeting and reduced offtumor toxicity.Emerging gene delivery technologies,including viral vectors,transposons,CRISPR/Cas9,and RNA-based electroporation,are improving CART production.Despite notable clinical success,particularly in CD19-and B-cell maturation antigen-targeted therapies,CAR-T applications face challenges,including cell exhaustion,antigen escape,and therapy-induced toxicities,such as cytokine release syndrome and neurotoxicity.Ongoing efforts in engineering innovation,clinical trials,and regulatory support continue to shape CAR-T therapy into a safer,more precise tool for cancer treatment.This review highlights current advances while outlining the barriers and future prospects of CAR-T immunotherapy.展开更多
Chimeric antigen receptor-T(CAR-T)cell therapy is a precise immunotherapy for lymphoma.However,its long-term efficacy faces many challenges related to tumor cell heterogeneity,interference from immunosuppressive micro...Chimeric antigen receptor-T(CAR-T)cell therapy is a precise immunotherapy for lymphoma.However,its long-term efficacy faces many challenges related to tumor cell heterogeneity,interference from immunosuppressive microenvironments,CAR-T cell exhaustion,and unmanageable adverse events.Diverse modifications have been introduced into conventional CAR-T cells to overcome these obstacles;examples include addition of recognition sites to prevent immune escape,coupling of cytokine domains to enhance killing ability,blocking of immune checkpoint signals to resist tumor microenvironments,and inclusion of suicide systems or safety switches to improve safety and flexibility.With increasing understanding of the importance of metabolism and epigenetics in cancer and cytotherapy,glycolysis,methylation,and acetylation have become crucial CAR-T cell therapeutic targets.Universal and in situ CAR-T cells are also expected to be used in clinical applications,thus providing hope to patients with relapsed/refractory lymphomas.展开更多
Background:Hepatocellular carcinoma(HCC)is a health problem due to multi-drug resistance(MDR).Codelivery of multiple oncotherapy in one cargo as chimeric cancer therapy(CCT)is suggested as a solution for MDR.This stud...Background:Hepatocellular carcinoma(HCC)is a health problem due to multi-drug resistance(MDR).Codelivery of multiple oncotherapy in one cargo as chimeric cancer therapy(CCT)is suggested as a solution for MDR.This study aims to engineer chitosan-coated nanostructure lipid carriers(NLCs)loaded with gefitinib(GF)and simvastatin(SV)as CCT for HCC.Methods:Both GF and SV-loaded nanostructure lipids carriers(GFSVNLC)and chitosan-capped GF and SV-loaded nanostructure lipids carriers(CGFSVNLC)formulations were assembled by topdown techniques.Moreover,particle size(PS),zeta potential(ZP),and polydispersity index(PDI)were measured by Zetasizer.The biosafety of GFSVNLC preparations was investigated by using erythrocytes as a biological model.The cytotoxic,and apoptotic effects of the prepared GFSVNLCs were investigated using HepG2 cell lines as a substitute model for HCC.The effect of GF,SV,and NLC composition on JNK3,HDAC6,and telomerase was studied using molecular docking simulation(MDS).Results:The present results revealed that the obtained GFSVNLC and CGFSVNLC have nanosized and consistent,CS coating shifts anionic ZP of GFSVNLC into CGFSVNLC with cationic ZP.Moreover,both formulations are biocompatible as indicated by their gentle effect on erythrocyte hemolysis.The treatment of HepG2 cells with GFSVNLC,and CGFSVNLC induced marked cell death compared to other groups with a decrease of IC50.Equally,the percentage of the apoptotic HepG2 cells was increased upon treatment of the cells with GFSV,GFSVNLC,and CGFSVNLC compared to the control group.Additionally,GF,SV,stearic acid(SA),and oleic acid(OA)modulate the activity of JNK3,HDAC6,and telomerase.Conclusions:This study suggests CGFSVNLC achieves codelivery,selective targeting,and enhancing the synergistic effect of GF and SV for inducing HepG2 cell death.Mechanistically,CGFSVNLC inhibits key cascades implicated in MDR and HepG2 cell survival.CGFSVNLC is promising for overcoming drug resistance mechanisms and improving therapeutic outcomes against HepG2 cells.展开更多
Chimeric antigen receptor natural killer(CAR-NK)cell therapy is an alternative immunotherapy that provides robust tumor-eliminating effects without inducing life-threatening toxicities and graft-versus-host disease.CA...Chimeric antigen receptor natural killer(CAR-NK)cell therapy is an alternative immunotherapy that provides robust tumor-eliminating effects without inducing life-threatening toxicities and graft-versus-host disease.CAR-NK cell therapy has enabled the development of“off-the-shelf”products that bypass the lengthy and expensive cell manufacturing process1.展开更多
The inherent low immunogenicity and immunosuppressive metabolism of solid tumors significantly attenuate the immunotherapeutic effect and restrict the immune response.In this work,an endoplasmic reticulum(ER)targeting...The inherent low immunogenicity and immunosuppressive metabolism of solid tumors significantly attenuate the immunotherapeutic effect and restrict the immune response.In this work,an endoplasmic reticulum(ER)targeting photodynamic oxidizer(designated as PhotoOx)is fabricated to boost the antitumor immunity by integrating photodynamic therapy(PDT)induced immunogenic cell death(ICD)with indoleamine 2,3-dioxygenase 1(IDO1)inhibition.Among which,an ER targeting photosensitizer-peptide conjugate called PhotoPe is rationally designed for optimal functionality and amphiphilicity,which could self-assemble into nano-micelles co-delivering chlorin e6 and NLG919.PhotoOx exhibits a good stability to enable ER targeting drug delivery,which could induce ER rupture to intensify PDT induced ICD and release damage associated molecular patterns(DAMPs).Furthermore,PhotoOx could effectively initiate immunological cascades,leading to the suppression of regulatory T cells(Tregs)and activation of CD8^(+)T cells when combines with IDO inhibition.Furthermore,the multi-synergistic effects of PhotoOx activate a robust systemic anti-tumor immune response,resulting in the eradication of lung and liver metastases.Such a medication strategy might inspire the rational design of biomedicine for precise drug delivery,which also provides a sophisticated mechanism for addressing the challenges of solid tumor treatment.展开更多
Glypican-3(GPC3)is a tumor-associated antigen that is specifically expressed in hepatocellular carcinoma(HCC)and having relatively low levels in normal tissues.This unique expression pattern positions GPC3 as a potent...Glypican-3(GPC3)is a tumor-associated antigen that is specifically expressed in hepatocellular carcinoma(HCC)and having relatively low levels in normal tissues.This unique expression pattern positions GPC3 as a potential target for precision therapy and drug development in HCC.Recent studies have shown significant advancements in GPC3-targeted therapies and immunotherapies,particularly for patients with advanced or treatment-resistant HCC.Although certain clinical trials have yielded suboptimal results,numerous ongoing studies continue to explore its therapeutic efficacy.This mini-review focuses on the latest research developments regarding GPC3 as a therapeutic target across various HCC treatment strategies,including monoclonal antibodies,bispecific antibodies,chimeric antigen receptor-T-cell therapies,and other innovative approaches.In addition,the limitations of GPC3-targeted therapies and their future application prospects in HCC treatment are discussed.The review particularly emphasizes the unmet need for future research directions,such as combination immuno-therapy strategies and novel drug designs.Through the integration of innovative technologies and clinical validation,GPC3 holds strong potential as a promising breakthrough in the treatment of HCC,offering new opportunities for enhancing patient outcomes and improving therapeutic efficacy.展开更多
Hepatocellular carcinoma(HCC)is the fourth leading cause of cancer-related mortality worldwide1.The primary treatment options for this disease are surgical resection and liver transplantation.Unfortunately,most HCC ca...Hepatocellular carcinoma(HCC)is the fourth leading cause of cancer-related mortality worldwide1.The primary treatment options for this disease are surgical resection and liver transplantation.Unfortunately,most HCC cases are diagnosed in advanced stages and are inoperable.Even after surgery,the long-term prognosis remains unsatisfactory,because of a high recurrence rate.展开更多
This review aims to explore the development,challenges,and future directions of UCAR cell therapy as a scalable alternative to autologous CAR-T for cancer treatment.Consequently,limitations of autologous CAR-T,includi...This review aims to explore the development,challenges,and future directions of UCAR cell therapy as a scalable alternative to autologous CAR-T for cancer treatment.Consequently,limitations of autologous CAR-T,including long production,variable quality,and cost,drive off-the-shelf UCAR development to standardize manufacturing and improve access.Current UCAR-T cell strategies focus on mitigating the risks of graft-vs.-host disease and host-vs.-graft rejection through advanced gene editing technologies,including clustered regularly interspaced short palindromic repeat-associated system Cas9-mediated knockout of the T cell receptor,human leukocyte antigen,and cluster of differentiation 52(CD52).Beyond conventional T cells,cell types such as double-negative T cells,γδT cells,and virus-specific T cells are being engineered with CARs to improve tumor targeting and minimize off-tumor toxicity.UCAR-T therapy is frequently used for hematologic malignancies,including acute lymphoblastic leukemia,non-Hodgkin lymphoma,andmultiplemyeloma,with efficacy and safety supported by numerous clinical studies.Although trials for solid tumors(e.g.,CYAD-101,CTX130)show modest responses,challenges such as tumor heterogeneity and T cell exhaustion remain.Future research should focus on optimizing gene editing precision,integrating combination therapies,and advancing scalable manufacturing platforms.With expanded targets and cell types,UCAR therapies show promise for both hematologic and solid tumors,reshaping cancer treatment and patient outcomes.展开更多
Autologous stem cell transplantation(ASCT)and chimeric antigen receptor T-cell(CAR-T)therapy represent pivotal treatments for hematologic malignancies,each with distinct strengths and limitations.ASCT reduces tumor bu...Autologous stem cell transplantation(ASCT)and chimeric antigen receptor T-cell(CAR-T)therapy represent pivotal treatments for hematologic malignancies,each with distinct strengths and limitations.ASCT reduces tumor burden through myeloablative conditioning but remains susceptible to relapse,while CAR-T therapy precisely targets malignant cells but encounters challenges,including cytokine release syndrome(CRS),immune effector cell-associated neurotoxicity syndrome(ICANS),and limited persistence.Emerging evidence suggests that combining ASCT with CAR-T therapy yields synergistic effects.ASCT reshapes the immune microenvironment,lowers immunosuppressive cells and CRS risk,while CAR-T eliminates residual disease and promotes immune recovery.Clinical trials in relapsed/refractory B-cell lymphomas and multiple myeloma demonstrate complete remission rates(CRR)of 72%-100%and two-year progression-free survival(PFS)rates of 59%-83%,with severe CRS/ICANS incidences below 10%.However,the precise mechanisms underlying this synergy,optimal timing of CAR-T infusion after ASCT,and ideal dosing regimens require further definition.Future research should prioritize large-scale,randomized controlled trials and establish standardized protocols for toxicity management to maximize therapeutic benefits.By integrating the complementary strengths of ASCT and CAR-T,this combination strategy represents a promising approach for improving outcomes in high-risk hematologic malignancies;however,additional studies are necessary to validate its efficacy and expand its clinical applicability.展开更多
Objective:Immune effector cell-associated neurotoxic syndrome(ICANS)is one of the most concerning complications because of its life-threatening trait during chimeric antigen receptor(CAR)T-cell therapy and requires sp...Objective:Immune effector cell-associated neurotoxic syndrome(ICANS)is one of the most concerning complications because of its life-threatening trait during chimeric antigen receptor(CAR)T-cell therapy and requires specialized management.We summarized the nursing procedures of CAR T-cell therapy for ICANS to provide some references for the standardized nursing care of ICANS.Methods:Based on previous experience and literature research,the professional team developed nursing procedures for ICANS for patients receiving CAR T-cell therapy.Results:The professional team developed step-by-step,sophisticated care procedures before,during,and after ICANS,including active communication and collaboration,knowledge updates,personalized nursing plans,advanced assessments,close monitoring of vital signs and laboratory tests,medication plans,timely and correct drug administration,dietary nursing interventions,safety care,nursing of other complications,follow-up of toxicities,and keeping an eye on the potential side effects.Conclusions:ICANS is unique,with varying clinical symptoms and rapid progression.Timely and accurate treatment and care of patients is an effective way to manage ICANS.Professional and detailed procedures are required to guide nurses to manage patients’ICANS,so as to improve the safety of patients and the promotion of CAR T-cell therapy.展开更多
Colorectal cancer(CRC)is the third most common cancer worldwide and remains a major treatment challenge,particularly in advanced and metastatic stages.Current standard treatments have limited efficacy,underscoring the...Colorectal cancer(CRC)is the third most common cancer worldwide and remains a major treatment challenge,particularly in advanced and metastatic stages.Current standard treatments have limited efficacy,underscoring the urgent need for innovative strategies.Adoptive cell therapy(ACT),which involves in vitro expansion or genetic engineering of immune cells,is a promising approach to bolster anti-tumor immune responses.Key ACT modalities include chimeric antigen receptor(CAR)T cells,tumor-infiltrating lymphocytes(TILs),and T cell receptor(TCR)-engineered T cells.CAR-T cell therapy has shown success in hematological malignancies but faces significant challenges in solid tumors like CRC.These challenges include antigen heterogeneity,an immunosuppressive tumor microenvironment,on-target off-tumor toxicity,among other factors.To address these limitations,combinatorial approaches,such as immune checkpoint inhibitors,cytokines,and advanced gene-editing tools like CRISPR/Cas9,are being actively explored.These strategies aim to enhance CAR-T cell specificity,improve resistance to immunosuppressive signals,and optimize in vivo functionality.This review summarizes ACT approaches for CRC,with a focus on CAR-T therapy.It briefly introduces TILs and TCR-T cells,while emphasizing the major challenges faced by CAR-T therapy in solid tumors and discusses potential strategies to improve therapeutic outcomes.展开更多
Objective Acute myeloid leukemia(AML)is a highly heterogeneous disease,and molecular events such as DNMT3A gene mutations are associated with poor prognosis in AML patients.Consequently,there is an urgent need for a n...Objective Acute myeloid leukemia(AML)is a highly heterogeneous disease,and molecular events such as DNMT3A gene mutations are associated with poor prognosis in AML patients.Consequently,there is an urgent need for a novel therapeutic approach for AML.Methods DNMT3A mRNA and protein expression were confirmed in DNMT3A-mutant AML cells via RT-qPCR and Western blotting.Cell proliferation and apoptosis were assessed via CCK-8 and Annexin V/PI staining,respectively.Flow cytometry was used to analyze surface antigens and CD44v6 CAR-T-cell transfection efficiency.CD44v6-directed CAR plasmids were constructed,and lentiviruses were packaged.Methylation-specific PCR was used to evaluate differences in promoter methylation,whereas ELISA was used to measure cytokine secretion.Results In this study,we found that the DNMT3A-mutant group presented significantly increased expression of CD44v6 on the cell surface.Methylation of the CD44 promoter region was lower in the mutant group than in the control group.CD44v6 CAR-T cells exhibited specific cytotoxicity against DNMT3A-mutant AML cells.Furthermore,pretreatment with low concentrations of decitabine significantly enhanced the killing effect of CD44v6 CAR-T cells on DNMT3A-mutant AML cells(P<0.05).Additionally,decitabine treatment upregulated the expression of CD44v6 on the surface of DNMT3A-mutant AML cells(P<0.05).Conclusion CD44v6 is a promising CAR-T-cell therapy target in AML patients with DNMT3A mutations.Notably,treatment with decitabine resulted in increased CD44v6 expression on the cell surface of DNMT3A-mutant AML cells.This increase in CD44v6 expression facilitates improved recognition and targeting by CD44v6 CAR-T cells.展开更多
Background:Chimeric antigen receptor T(CAR-T)cell therapies have demonstrated significant clinical efficacy in hematological malignancies.However,their application to solid tumors remains substantially limited by mult...Background:Chimeric antigen receptor T(CAR-T)cell therapies have demonstrated significant clinical efficacy in hematological malignancies.However,their application to solid tumors remains substantially limited by multiple challenges,including the risk of off-target effects.Hence,optimizing CAR-T cells for stronger antigen binding is essential.Methods:In this study,we employed a classical anti-human endothelial growth factor receptor 2(HER2)single-chain variable fragment(scFv)derived from trastuzumab,alongside an anti-HER2-13 scFv identified from a combinatorial cellular CAR library,for the construction of a third-generation CAR-T cell.Meanwhile,the phenotypes and both in vitro and in vivo functions of CAR-T cells transduced with the two scFvs via PiggyBac transposon-mediated gene transfer were compared.Results:The optimal ratio between the PiggyBac HER2-CAR-puro transposon and the Super PiggyBac transposase plasmid differed during the construction of the two HER2-targeted CAR-T cell types.The expansion abilities,CD3^(+)CAR^(+)population,CD4^(+)CAR^(+)/CD8^(+)CAR^(+)proportions,and memory and exhaustion markers between the two CAR-T groups were similar after using the optimized proportion of plasmid.Both CAR-T cell types exhibited significant antitumor activity,with the anti-HER2-13 CAR-T cells demonstrating superior target specificity.Therapeutic effects were observed with both CAR-T cells and trastuzumab in theMDA-MB-231HER2+breast tumor xenograft model,with anti-HER2-13 CAR-T cells demonstrating slightly enhanced efficacy and no evident offtarget toxicity.Conclusion:These results highlight the potential of anti-HER2-13 CAR-T cells to serve as a safer and more efficacious alternative in HER2-targeted therapy.展开更多
文摘A recently published study(Xin et al.,Prog Biochem Biophys,2026,53(2):431-441.DOI:10.3724/j.pibb.2025.0508)addresses the therapeutic challenges of pancreatic ductal adenocarcinoma(PDAC)by innovatively developing an orally administered nanogene delivery system.Designed to achieve in situ,efficient delivery of chimeric antigen receptor(CAR)genes to tumor sites,this approach offers a novel strategy for CAR-macrophage(CAR-M)based immunotherapy.Its key highlights are as follows.
基金supported by Key Research and Development Plan of Hunan Province(2024DK2006)the Fundamental Research Funds for the Central Universities of Central South University(1053320221769)+2 种基金Hunan Provincial Respiratory Disease Rehabilitation and Nursing Engineering Research Center Innovation Capacity Building Project(No.202012)the Zhangjiajie Science and Technology Development Key Special Project(No.202304)the National Key Clinical Specialty Major Scientific Research Project(No.20230382).
文摘Background:Glioblastoma(GBM)prognosis has seen little improvement over the past two decades.While immunotherapy has revolutionized cancer treatment,its impact on GBM remains limited.To characterize the evolving research landscape and identify future directions in GBM immunotherapy,we conducted a comprehensive bibliometric review.Methods:All literature related to immunotherapy in GBM from 1999 to 2024 was collected from the Web of Science Core Collection.CtieSpace and VOSviewer were used to conduct bibliometric analysis and visualize the data.Results:Bibliometric analysis identified 5038 publications authored by 23,335 researchers from 4699 institutions across 96 countries/regions,published in 945 journals.The United States produced the highest number of publications,while Switzerland achieved the highest average citation rate.Duke University led in institutional output and citations.John H Sampson was the most productive author,and Roger Stupp was the most cited.Frontiers in Immunology published the most papers,while Clinical Cancer Research was the most cited journal.Research focus centered on adoptive T cell therapy,particularly chimeric antigen receptor(CAR)-T cells with 572 dedicated publications.Within CAR-T research for GBM,the University of Pennsylvania was the leading institution,Frontiers in Immunology the predominant journal,and Christine E Brown(City of Hope National Medical Center)was the most prolific and cited author.Conclusions:There has been a growing interest in GBM immunotherapy over past decades.The United States is the dominant contributor.CAR-T therapy represents the primary research focus.Emerging strategies like chimeric antigen receptor-modified natural killer(CAR-NK)cells,chimeric antigen receptor-engineered macrophages(CAR-M),and cytomegalovirus-specific T cell receptor(CMV-TCR)T cells are gaining prominence,aiming to address limitations in antigen recognition inherent to CAR-T therapy for GBM.
基金supported by the Kazan Federal University Strategic Academic Leadership Program(PRIORITY-2030).
文摘Classical Hodgkin lymphoma(cHL)is characterized by rare Hodgkin/Reed-Sternberg(HRS)tumor cells that uniformly express cluster of differentiation(CD)30 molecules and orchestrate an immunosuppressive tumor microenvironment,making CD30 an attractive and selective therapeutic target.We summarize the biological rationale for CD30 as a therapeutic target and the preclinical and clinical evidence across major platforms:antibody-drug conjugates(brentuximab vedotin),monoclonal antibodies(including acimtamig and its combinations with Natural Killer cells),second-and third-generation chimeric antigen receptor(CAR)-T cells,and alternative modalities.Particular attention is given to standardized response assessment(IWG,Lugano,RECIL criteria),which enables appropriate cross-trial comparisons.Taken together,the data indicate that beyond the established role of brentuximab vedotin,CD30-directed CAR-T cells and bispecific antibodies demonstrate high activity in refractory cHL,especially when used with fludarabine-containing lymphodepletion,combined with programmed cell death 1(PD-1)receptor blockade as a strategy to eradicate minimal residual disease.Key challenges include durable effector-cell persistence and optimization of sequencing and combinations;notably,loss of CD30 as an escape mechanism appears uncommon.Integrating mechanistic insights into HRS biology with clinical trial data highlights strategies to enhance the efficacy,safety,and accessibility of CD30-directed immunotherapy.This review aims to provide a concise overview of CD30-targeted approaches in cHL,emphasizing therapeutic outcomes and the evolution of CAR-T technologies.
基金Ministry of Science and Technology of China(STI2030-Major Projects),No.2022ZD0204700(to WW)the National Natural Science Foundation of China,No.82301572(to XZ)China Postdoctoral Science Foundation,No.2023M731202(to XZ).
文摘In the human spinal cord,astrocytes are the major glial cells.In vitro studies of human astrocytes are relatively simple.However,the straightforward nature of the in vitro environment and complex nature of the in vivo environment limit comprehensive investigations into the structure and function of human astrocytes.Additionally,in vivo studies of human astrocytes are further limited by ethical issues.This means there is an urgent need to develop effective in vivo models to study the structure and function of human astrocytes.Here,we first directed human embryonic stem cells to differentiate into human spinal cord dorsal neural stem/progenitor cells in vitro,before transplanting these cells into the gray matter of the cervical spinal cord(C5-T2 segments)of naïve nude rats to create a chimeric human astrocytic rat spinal cord model.The transplanted human spinal cord dorsal neural stem/progenitor cells survived for at least 20 months in the spinal cord environment of the rats,with over 90%differentiating into human astrocytes.These human astrocytes were able to migrate caudally for long distances along the white matter towards the spinal cord.They expressed astrocytic cytoskeletal proteins and functionally-related proteins,suggesting their maturation and structural integration into the rat spinal cord.Thus,this humanized astrocyte chimeric rat spinal cord model provides a valuable tool for studying the role of human spinal cord astrocytes in various spinal diseases.
基金supported by the National Research Foundation of Korea(NRF)through the Ministry of Education(2021R1I1A3059820)(to Jea-Hyun Baek).
文摘The paradigm of cancer treatment has been reshaped by chimeric antigen receptor(CAR)αβT cell therapy,yet its full potential remains constrained by fundamental limitations.While conventional CARαβT cells have achieved notable success in hematological malignancies,their broader application is hindered by the high cost and delays of autologous manufacturing,as well as the critical risk of graft-vs-host disease(GvHD).In addition,their efficacy against solid tumors is often compromised by the immunosuppressive tumor microenvironment(TME).As a promising solution,γδT cells are being developed as an alternative CAR platform.Their intrinsic ability to recognize transformed cells in a major histocompatibility complex(MHC)-independent manner minimizes the risk of GvHD and supports the creation of safe,effective allogeneic therapies.Building on this unique biology,the therapeutic efficacy of CARγδT cells is being enhanced through advanced engineering strategies.Key innovations include“armoring”technologies,such as cytokine secretion,checkpoint blockade,and metabolic rewiring,to overcome local immunosuppression and improve persistence,as well as the use of induced pluripotent stem cells(iPSCs)to generate standardized products from a renewable and consistent source.This expanding technological toolbox is also enabling novel applications beyond oncology.For example,chimeric autoantibody receptor(CAAR)constructs built onγδT cells integrate both classical and emerging insights into CARγδT cell therapy,highlighting innovations that are driving the field toward safer,more versatile,and longer-lasting treatments for cancer and autoimmunity.In light of these advancements,this review provides an overview of the current understanding ofγδT cell biology and highlights emerging engineering strategies that enhance the efficacy and durability of CARγδT cells across oncologic and autoimmune contexts.
文摘Background:In patients with autoimmune hemolytic anemia(AIHA),the risk of relapse is high owing to persistent autoreactive B-cell activity.Multirefractory AIHA is a more advanced stage of disease that is defined by a lack of response to at least three lines of therapy.CD19-directed chimeric antigen receptor(CAR)T-cell therapy results in profound B-cell depletion and may be a useful approach to achieving drug-free remission in multirefractory AIHA.
文摘Chimeric antigen receptor T(CAR-T)cell therapy represents a major advance in cancer immunotherapy,offering targeted treatment options,particularly for hematologic malignancies.This review comprehensively explores the structural evolution,production processes,and cytotoxic mechanisms underlying CAR-T function.Therapy involves engineering autologous T cells with synthetic receptors that allow major histocompatibility complex-independent recognition of tumor-associated antigens.Key structural components such as antigen recognition domains,spacers,transmembrane,and intracellular domains are optimized to enhance specificity,persistence,and cytotoxicity.CAR-T therapy exerts antitumor effects via granzyme-perforin degranulation,Fas/Fas ligand signaling,and cytokine secretion.Over time,the development of second-to fifth-generation CARs has incorporated costimulatory molecules,transcriptional regulation,and logic-gated control to improve efficacy and safety.Additionally,novel engineering strategies such as dual CARs,tandem CARs,SynNotch systems,and universal or inhibitory CARs have expanded antigen targeting and reduced offtumor toxicity.Emerging gene delivery technologies,including viral vectors,transposons,CRISPR/Cas9,and RNA-based electroporation,are improving CART production.Despite notable clinical success,particularly in CD19-and B-cell maturation antigen-targeted therapies,CAR-T applications face challenges,including cell exhaustion,antigen escape,and therapy-induced toxicities,such as cytokine release syndrome and neurotoxicity.Ongoing efforts in engineering innovation,clinical trials,and regulatory support continue to shape CAR-T therapy into a safer,more precise tool for cancer treatment.This review highlights current advances while outlining the barriers and future prospects of CAR-T immunotherapy.
基金supported by grants from the Science Technology Department of Zhejiang Province(Grant No.2021C03117)Noncommunicable Chronic DiseasesNational Science and Technology Major Project(Grant No.2023ZD0501300)+1 种基金National Natural Science Foundation of China(Grant No.82170219)Zhejiang Provincial Natural Science Foundation of China(Grant No.LQ24H080009)。
文摘Chimeric antigen receptor-T(CAR-T)cell therapy is a precise immunotherapy for lymphoma.However,its long-term efficacy faces many challenges related to tumor cell heterogeneity,interference from immunosuppressive microenvironments,CAR-T cell exhaustion,and unmanageable adverse events.Diverse modifications have been introduced into conventional CAR-T cells to overcome these obstacles;examples include addition of recognition sites to prevent immune escape,coupling of cytokine domains to enhance killing ability,blocking of immune checkpoint signals to resist tumor microenvironments,and inclusion of suicide systems or safety switches to improve safety and flexibility.With increasing understanding of the importance of metabolism and epigenetics in cancer and cytotherapy,glycolysis,methylation,and acetylation have become crucial CAR-T cell therapeutic targets.Universal and in situ CAR-T cells are also expected to be used in clinical applications,thus providing hope to patients with relapsed/refractory lymphomas.
基金Support Project(RSPD2024R1037),King Saud University,Riyadh,Saudi Arabia.
文摘Background:Hepatocellular carcinoma(HCC)is a health problem due to multi-drug resistance(MDR).Codelivery of multiple oncotherapy in one cargo as chimeric cancer therapy(CCT)is suggested as a solution for MDR.This study aims to engineer chitosan-coated nanostructure lipid carriers(NLCs)loaded with gefitinib(GF)and simvastatin(SV)as CCT for HCC.Methods:Both GF and SV-loaded nanostructure lipids carriers(GFSVNLC)and chitosan-capped GF and SV-loaded nanostructure lipids carriers(CGFSVNLC)formulations were assembled by topdown techniques.Moreover,particle size(PS),zeta potential(ZP),and polydispersity index(PDI)were measured by Zetasizer.The biosafety of GFSVNLC preparations was investigated by using erythrocytes as a biological model.The cytotoxic,and apoptotic effects of the prepared GFSVNLCs were investigated using HepG2 cell lines as a substitute model for HCC.The effect of GF,SV,and NLC composition on JNK3,HDAC6,and telomerase was studied using molecular docking simulation(MDS).Results:The present results revealed that the obtained GFSVNLC and CGFSVNLC have nanosized and consistent,CS coating shifts anionic ZP of GFSVNLC into CGFSVNLC with cationic ZP.Moreover,both formulations are biocompatible as indicated by their gentle effect on erythrocyte hemolysis.The treatment of HepG2 cells with GFSVNLC,and CGFSVNLC induced marked cell death compared to other groups with a decrease of IC50.Equally,the percentage of the apoptotic HepG2 cells was increased upon treatment of the cells with GFSV,GFSVNLC,and CGFSVNLC compared to the control group.Additionally,GF,SV,stearic acid(SA),and oleic acid(OA)modulate the activity of JNK3,HDAC6,and telomerase.Conclusions:This study suggests CGFSVNLC achieves codelivery,selective targeting,and enhancing the synergistic effect of GF and SV for inducing HepG2 cell death.Mechanistically,CGFSVNLC inhibits key cascades implicated in MDR and HepG2 cell survival.CGFSVNLC is promising for overcoming drug resistance mechanisms and improving therapeutic outcomes against HepG2 cells.
基金supported by grants from the Noncommunicable Chronic Diseases-National Science and Technology Major Project(Grant No.2023ZD0501300)Science Technology Department of Zhejiang Province(Grant No.2021C03117)+2 种基金National Natural Science Foundation of China(Grant No.82350104 and 82170219)Natural Science Foundation of Zhejiang Province,China(Grant No.LY23H080004 and LY24H080001)Medical Health Science and Technology Project of Zhejiang Provincial Health Commission(Grant No.2021KY199)。
文摘Chimeric antigen receptor natural killer(CAR-NK)cell therapy is an alternative immunotherapy that provides robust tumor-eliminating effects without inducing life-threatening toxicities and graft-versus-host disease.CAR-NK cell therapy has enabled the development of“off-the-shelf”products that bypass the lengthy and expensive cell manufacturing process1.
基金supported by the National Natural Science Foundation of China(No.32371394)the National Key R&D Program of China(No.2021YFD1800600)+3 种基金the Special Projects in Key Areas of Colleges and Universities in Guangdong Province(No.2022ZDZX2046)the Open Project of State Key Laboratory of Respiratory Disease(No.SKLRD-OP-202502)the Postdoctoral Fellowship Program of CPSF(No.GZC20230619)Postdoctoral startup funding of Guangzhou Medical University(No.Q0301–130)。
文摘The inherent low immunogenicity and immunosuppressive metabolism of solid tumors significantly attenuate the immunotherapeutic effect and restrict the immune response.In this work,an endoplasmic reticulum(ER)targeting photodynamic oxidizer(designated as PhotoOx)is fabricated to boost the antitumor immunity by integrating photodynamic therapy(PDT)induced immunogenic cell death(ICD)with indoleamine 2,3-dioxygenase 1(IDO1)inhibition.Among which,an ER targeting photosensitizer-peptide conjugate called PhotoPe is rationally designed for optimal functionality and amphiphilicity,which could self-assemble into nano-micelles co-delivering chlorin e6 and NLG919.PhotoOx exhibits a good stability to enable ER targeting drug delivery,which could induce ER rupture to intensify PDT induced ICD and release damage associated molecular patterns(DAMPs).Furthermore,PhotoOx could effectively initiate immunological cascades,leading to the suppression of regulatory T cells(Tregs)and activation of CD8^(+)T cells when combines with IDO inhibition.Furthermore,the multi-synergistic effects of PhotoOx activate a robust systemic anti-tumor immune response,resulting in the eradication of lung and liver metastases.Such a medication strategy might inspire the rational design of biomedicine for precise drug delivery,which also provides a sophisticated mechanism for addressing the challenges of solid tumor treatment.
文摘Glypican-3(GPC3)is a tumor-associated antigen that is specifically expressed in hepatocellular carcinoma(HCC)and having relatively low levels in normal tissues.This unique expression pattern positions GPC3 as a potential target for precision therapy and drug development in HCC.Recent studies have shown significant advancements in GPC3-targeted therapies and immunotherapies,particularly for patients with advanced or treatment-resistant HCC.Although certain clinical trials have yielded suboptimal results,numerous ongoing studies continue to explore its therapeutic efficacy.This mini-review focuses on the latest research developments regarding GPC3 as a therapeutic target across various HCC treatment strategies,including monoclonal antibodies,bispecific antibodies,chimeric antigen receptor-T-cell therapies,and other innovative approaches.In addition,the limitations of GPC3-targeted therapies and their future application prospects in HCC treatment are discussed.The review particularly emphasizes the unmet need for future research directions,such as combination immuno-therapy strategies and novel drug designs.Through the integration of innovative technologies and clinical validation,GPC3 holds strong potential as a promising breakthrough in the treatment of HCC,offering new opportunities for enhancing patient outcomes and improving therapeutic efficacy.
基金supported by the RGC Research Impact Fund(Grant No.R5008-22F).
文摘Hepatocellular carcinoma(HCC)is the fourth leading cause of cancer-related mortality worldwide1.The primary treatment options for this disease are surgical resection and liver transplantation.Unfortunately,most HCC cases are diagnosed in advanced stages and are inoperable.Even after surgery,the long-term prognosis remains unsatisfactory,because of a high recurrence rate.
文摘This review aims to explore the development,challenges,and future directions of UCAR cell therapy as a scalable alternative to autologous CAR-T for cancer treatment.Consequently,limitations of autologous CAR-T,including long production,variable quality,and cost,drive off-the-shelf UCAR development to standardize manufacturing and improve access.Current UCAR-T cell strategies focus on mitigating the risks of graft-vs.-host disease and host-vs.-graft rejection through advanced gene editing technologies,including clustered regularly interspaced short palindromic repeat-associated system Cas9-mediated knockout of the T cell receptor,human leukocyte antigen,and cluster of differentiation 52(CD52).Beyond conventional T cells,cell types such as double-negative T cells,γδT cells,and virus-specific T cells are being engineered with CARs to improve tumor targeting and minimize off-tumor toxicity.UCAR-T therapy is frequently used for hematologic malignancies,including acute lymphoblastic leukemia,non-Hodgkin lymphoma,andmultiplemyeloma,with efficacy and safety supported by numerous clinical studies.Although trials for solid tumors(e.g.,CYAD-101,CTX130)show modest responses,challenges such as tumor heterogeneity and T cell exhaustion remain.Future research should focus on optimizing gene editing precision,integrating combination therapies,and advancing scalable manufacturing platforms.With expanded targets and cell types,UCAR therapies show promise for both hematologic and solid tumors,reshaping cancer treatment and patient outcomes.
基金supported by funding from the National KeyR&D Program of China(No.2022YFC2502604)the National Natural Science Foundation of China(No.82470194)+1 种基金the CAMS Innovation Fund for Medical Sciences(CIFMS)(No.2024-I2M-3-021)the ChenXiaoping Foundation for the development of Science and Technology of Hubei Province(No.CXPJJH122001-2221)。
文摘Autologous stem cell transplantation(ASCT)and chimeric antigen receptor T-cell(CAR-T)therapy represent pivotal treatments for hematologic malignancies,each with distinct strengths and limitations.ASCT reduces tumor burden through myeloablative conditioning but remains susceptible to relapse,while CAR-T therapy precisely targets malignant cells but encounters challenges,including cytokine release syndrome(CRS),immune effector cell-associated neurotoxicity syndrome(ICANS),and limited persistence.Emerging evidence suggests that combining ASCT with CAR-T therapy yields synergistic effects.ASCT reshapes the immune microenvironment,lowers immunosuppressive cells and CRS risk,while CAR-T eliminates residual disease and promotes immune recovery.Clinical trials in relapsed/refractory B-cell lymphomas and multiple myeloma demonstrate complete remission rates(CRR)of 72%-100%and two-year progression-free survival(PFS)rates of 59%-83%,with severe CRS/ICANS incidences below 10%.However,the precise mechanisms underlying this synergy,optimal timing of CAR-T infusion after ASCT,and ideal dosing regimens require further definition.Future research should prioritize large-scale,randomized controlled trials and establish standardized protocols for toxicity management to maximize therapeutic benefits.By integrating the complementary strengths of ASCT and CAR-T,this combination strategy represents a promising approach for improving outcomes in high-risk hematologic malignancies;however,additional studies are necessary to validate its efficacy and expand its clinical applicability.
基金supported by the Special Research Fund for Central Universities,Peking Union Medical College(No.3332023063).
文摘Objective:Immune effector cell-associated neurotoxic syndrome(ICANS)is one of the most concerning complications because of its life-threatening trait during chimeric antigen receptor(CAR)T-cell therapy and requires specialized management.We summarized the nursing procedures of CAR T-cell therapy for ICANS to provide some references for the standardized nursing care of ICANS.Methods:Based on previous experience and literature research,the professional team developed nursing procedures for ICANS for patients receiving CAR T-cell therapy.Results:The professional team developed step-by-step,sophisticated care procedures before,during,and after ICANS,including active communication and collaboration,knowledge updates,personalized nursing plans,advanced assessments,close monitoring of vital signs and laboratory tests,medication plans,timely and correct drug administration,dietary nursing interventions,safety care,nursing of other complications,follow-up of toxicities,and keeping an eye on the potential side effects.Conclusions:ICANS is unique,with varying clinical symptoms and rapid progression.Timely and accurate treatment and care of patients is an effective way to manage ICANS.Professional and detailed procedures are required to guide nurses to manage patients’ICANS,so as to improve the safety of patients and the promotion of CAR T-cell therapy.
基金Supported by the Natural Science Foundation of the Science and Technology Commission of Shanghai Municipality,China,No.23ZR1458300Key Discipline Project of Shanghai Municipal Health System,China,No.2024ZDXK0004+1 种基金Doctoral Innovation Talent Base Project for Diagnosis and Treatment of Chronic Liver Diseases,China,No.RCJD2021B02Pujiang Project of Shanghai Magnolia Talent Plan,China,No.24PJD098.
文摘Colorectal cancer(CRC)is the third most common cancer worldwide and remains a major treatment challenge,particularly in advanced and metastatic stages.Current standard treatments have limited efficacy,underscoring the urgent need for innovative strategies.Adoptive cell therapy(ACT),which involves in vitro expansion or genetic engineering of immune cells,is a promising approach to bolster anti-tumor immune responses.Key ACT modalities include chimeric antigen receptor(CAR)T cells,tumor-infiltrating lymphocytes(TILs),and T cell receptor(TCR)-engineered T cells.CAR-T cell therapy has shown success in hematological malignancies but faces significant challenges in solid tumors like CRC.These challenges include antigen heterogeneity,an immunosuppressive tumor microenvironment,on-target off-tumor toxicity,among other factors.To address these limitations,combinatorial approaches,such as immune checkpoint inhibitors,cytokines,and advanced gene-editing tools like CRISPR/Cas9,are being actively explored.These strategies aim to enhance CAR-T cell specificity,improve resistance to immunosuppressive signals,and optimize in vivo functionality.This review summarizes ACT approaches for CRC,with a focus on CAR-T therapy.It briefly introduces TILs and TCR-T cells,while emphasizing the major challenges faced by CAR-T therapy in solid tumors and discusses potential strategies to improve therapeutic outcomes.
基金supported by the National Natural Science Foundation of China(No.82270177)Chen Xiaoping Foundation for the Development of Science and Technology of Hubei Province(CXPJJH122001-2221).
文摘Objective Acute myeloid leukemia(AML)is a highly heterogeneous disease,and molecular events such as DNMT3A gene mutations are associated with poor prognosis in AML patients.Consequently,there is an urgent need for a novel therapeutic approach for AML.Methods DNMT3A mRNA and protein expression were confirmed in DNMT3A-mutant AML cells via RT-qPCR and Western blotting.Cell proliferation and apoptosis were assessed via CCK-8 and Annexin V/PI staining,respectively.Flow cytometry was used to analyze surface antigens and CD44v6 CAR-T-cell transfection efficiency.CD44v6-directed CAR plasmids were constructed,and lentiviruses were packaged.Methylation-specific PCR was used to evaluate differences in promoter methylation,whereas ELISA was used to measure cytokine secretion.Results In this study,we found that the DNMT3A-mutant group presented significantly increased expression of CD44v6 on the cell surface.Methylation of the CD44 promoter region was lower in the mutant group than in the control group.CD44v6 CAR-T cells exhibited specific cytotoxicity against DNMT3A-mutant AML cells.Furthermore,pretreatment with low concentrations of decitabine significantly enhanced the killing effect of CD44v6 CAR-T cells on DNMT3A-mutant AML cells(P<0.05).Additionally,decitabine treatment upregulated the expression of CD44v6 on the surface of DNMT3A-mutant AML cells(P<0.05).Conclusion CD44v6 is a promising CAR-T-cell therapy target in AML patients with DNMT3A mutations.Notably,treatment with decitabine resulted in increased CD44v6 expression on the cell surface of DNMT3A-mutant AML cells.This increase in CD44v6 expression facilitates improved recognition and targeting by CD44v6 CAR-T cells.
基金supported by the Yunnan Fundamental Research Projects,China(Grant No.202201AS070068)Central Funds Guiding the Local Science and Technology Development,China(202207AB110017)+1 种基金The Science and Technology Fund of Kunming City,China(No.2019-1-N-25318000002027)The Scientific and Technological Innovation Team in Kunming Medical University,China(CXTD202215).
文摘Background:Chimeric antigen receptor T(CAR-T)cell therapies have demonstrated significant clinical efficacy in hematological malignancies.However,their application to solid tumors remains substantially limited by multiple challenges,including the risk of off-target effects.Hence,optimizing CAR-T cells for stronger antigen binding is essential.Methods:In this study,we employed a classical anti-human endothelial growth factor receptor 2(HER2)single-chain variable fragment(scFv)derived from trastuzumab,alongside an anti-HER2-13 scFv identified from a combinatorial cellular CAR library,for the construction of a third-generation CAR-T cell.Meanwhile,the phenotypes and both in vitro and in vivo functions of CAR-T cells transduced with the two scFvs via PiggyBac transposon-mediated gene transfer were compared.Results:The optimal ratio between the PiggyBac HER2-CAR-puro transposon and the Super PiggyBac transposase plasmid differed during the construction of the two HER2-targeted CAR-T cell types.The expansion abilities,CD3^(+)CAR^(+)population,CD4^(+)CAR^(+)/CD8^(+)CAR^(+)proportions,and memory and exhaustion markers between the two CAR-T groups were similar after using the optimized proportion of plasmid.Both CAR-T cell types exhibited significant antitumor activity,with the anti-HER2-13 CAR-T cells demonstrating superior target specificity.Therapeutic effects were observed with both CAR-T cells and trastuzumab in theMDA-MB-231HER2+breast tumor xenograft model,with anti-HER2-13 CAR-T cells demonstrating slightly enhanced efficacy and no evident offtarget toxicity.Conclusion:These results highlight the potential of anti-HER2-13 CAR-T cells to serve as a safer and more efficacious alternative in HER2-targeted therapy.
