Double-negative T cells:a promising avenue of adoptive cell therapy in transplant oncology

Tumor recurrence is one of the major life-threatening complications after liver transplantation for liver cancer.In addition to the common mechanisms underlying tumor recurrence,another unavoidable problem is that the immunosuppressive therapeutic regimen after transplantation could promote tumor recurrence and metastasis.Transplant oncology is an emerging field that addresses oncological challenges in transplantation.In this context,a comprehensive therapeutic management approach is required to balance the anti-tumor treatment and immunosuppressive status of recipients.Double-negative T cells(DNTs)are a cluster of heterogeneous cells mainly consisting of two subsets stratified by T cell receptor(TCR)type.Among them,TCRαβ^(+)DNTs are considered to induce immune suppression in immune-mediated diseases,while TCRγδ^(+)DNTs are widely recognized as tumor killers.As a composite cell therapy,healthy donor-derived DNTs can be propagated to therapeutic numbers in vitro and applied for the treatment of several malignancies without impairing normal tissues or being rejected by the host.In this work,we summarized the biological characteristics and functions of DNTs in oncology,immunology,and transplantation.Based on the multiple roles of DNTs,we propose that a new balance could be achieved in liver transplant oncology using them as an off-the-shelf adoptive cell therapy(ACT).

Ginsenoside Rg1 improves anti-tumor efficacy of adoptive cell therapy by enhancing T cell effector functions

Adoptive cell therapy(ACT)has emerged with remarkable efficacies for tumor immunotherapy.Chimeric antigen receptor(CAR)T cell therapy,as one of most promising ACTs,has achieved prominent effects in treating malignant hematological tumors.However,the insufficient killing activity and limited persistence of T cells in the immunosuppressive tumor microenvironment limit the further application of ACTs for cancer patients.Many studies have focused on improving cytotoxicity and persistence of T cells to achieve improved therapeutic effects.In this study,we explored the potential function in ACT of ginsenoside Rg1,the main pharmacologically active component of ginseng.We introduced Rg1 during the in vitro activation and expansion phase of T cells,and found that Rg1 treatment upregulated two T cell activation markers,CD69 and CD25,while promoting T cell differentiation towards a mature state.Transcriptome sequencing revealed that Rg1 influenced T cell metabolic reprogramming by strengthening mitochondrial biosynthesis.When co-cultured with tumor cells,Rg1-treated T cells showed stronger cytotoxicity than untreated cells.Moreover,adding Rg1 to the culture endowed CAR-T cells with enhanced anti-tumor efficacy.This study suggests that ginsenoside Rg1 provides a potential approach for improving the anti-tumor efficacy of ACT by enhancing T cell effector functions.

When immunotherapy meets liver transplantation for hepatocellular carcinoma: A bumpy but promising road

Liver transplantation(LT)is a highly curative therapy for patients with hepatocellular carcinoma(HCC).However,due to the shortage of donor livers and rapid progression of HCC,a majority of patients are dropped out from the waitlist.Recently,immunotherapy has shown great promise in the treatment of advanced HCC.However,the use of immunotherapy is limited in LT mainly due to the potentially increasing risk of graft rejection.One of the main challenges for researchers is the protection of donor graft from an immunotherapy-boosted immune response mounted by the host.Besides,the safety,availability,and costs of immunotherapy are other challenges that need to be addressed.Here,we reviewed the literature involving patients who received immunotherapy prior to transplant to avoid waitlist dropouts and following transplantation to prevent the progression of tumor recurrence and metastasis.Statistically,the incidence of rejection was 25.0%pre-transplant and 18.5%post-transplant.Based on the review of these clinical studies,we can conclude that conducting clinical trials on the safety and efficacy of currently available immunotherapy drugs and identifying novel immunotherapy targets through extensive research may be promising for patients who do not meet the selection criteria for LT and who experience post-transplant recurrence.To date,the clinical experience on the use of immunotherapy before or after LT comes from individual case studies.Although some of the reported results are promising,they are not sufficient to support the standardized use of immunotherapy in clinical practice.

Coating with flexible DNA network enhanced T-cell activation and tumor killing for adoptive cell therapy

Adoptive cell therapy(ACT)is an emerging powerful cancer immunotherapy,which includes a complex process of genetic modification,stimulation and expansion.During these in vitro or ex vivo manipulation,sensitive cells are inescapability subjected to harmful external stimuli.Although a variety of cytoprotection strategies have been developed,their application on ACT remains challenging.Herein,a DNA network is constructed on cell surface by rolling circle amplification(RCA),and T cell-targeted trivalent tetrahedral DNA nanostructure is used as a rigid scaffold to achieve high-efficient and selective coating for T cells.The cytoprotective DNA network on T-cell surface makes them aggregate over time to form cell clusters,which exhibit more resistance to external stimuli and enhanced activities in human peripheral blood mononuclear cells and liver cancer organoid killing model.Overall,this work provides a novel strategy for in vitro T cell-selective protection,which has a great potential for application in ACT.

Targets of immunotherapy for hepatocellular carcinoma: An update

Hepatocellular carcinoma,the most common primary liver cancer,in an immunogenic tumor with a poor prognosis because these tumors are diagnosed at late stages.Although,surgical resection,ablation,liver transplant,and locoregional therapies are available for early stages;however,there are yet no effective treatment for advanced and recurrent tumors.Immune checkpoint inhibitor therapy and adoptive cell transfer therapy has gained the popularity with some positive results because these therapies overcome anergy and systemic immune suppression.However,still there is a lack of an effective treatment and thus there is an unmet need of a novel treatment.At present,the focus of the research is on oncolytic viral therapy and combination therapy where therapies including radiotherapy,immune checkpoint therapy,adoptive cell transfer therapy,and vaccines are combined to get an additive or synergistic effect enhancing the immune response of the liver with a cytotoxic effect on tumor cells.This review discusses the recent key development,the basis of drug resistance,immune evasion,immune tolerance,the available therapies based on stage of the tumor,and the ongoing clinical trials on immune checkpoint inhibitor therapy,adoptive cell transfer therapy,oncolytic viral vaccine therapy,and combination therapy.

Tumor neoantigens: Novel strategies for application of cancer immunotherapy

Neoantigen-targeted immunotherapy is a rapidly advancing field that holds great promise for treating cancer.The recognition of antigens by immune cells is a crucial step in tumor-specific killing,and neoantigens generated by mutations in cancer cells possess high immunogenicity and are selectively expressed in tumor cells,making them an attractive therapeutic target.Currently,neoantigens find utility in various domains,primarily in the realm of neoantigen vaccines such as DC vaccines,nucleic acid vaccines,and synthetic long peptide vaccines.Additionally,they hold promise in adoptive cell therapy,encompassing tumor-infiltrating cells,T cell receptors,and chimeric antigen receptors which are expressed by genetically modified T cells.In this review,we summarized recent progress in the clinical use of tumor vaccines and adoptive cell therapy targeting neoantigens,discussed the potential of neoantigen burden as an immune checkpoint in clinical settings.With the aid of state-of-the-art sequencing and bioinformatics technologies,together with significant advancements in artificial intelligence,we anticipated that neoantigens will be fully exploited for personalized tumor immunotherapy,from screening to clinical application.

Evaluation of the safety and efficiency of cytotoxic T cell therapy sensitized by tumor antigens original from T-ALL-iPSC in vivo

Background:Since RNA sequencing has shown that induced pluripotent stem cells(iPSCs)share a common antigen profile with tumor cells,cancer vaccines that focus on iPSCs have made promising progress in recent years.Previously,we showed that iPSCs derived from leukemic cells of patients with primary T cell acute lymphoblastic leukemia(T-ALL)have a gene expression profile similar to that of T-ALL cell lines.Methods:Mice with T-ALL were treated with dendritic and T(DC-T)cells loaded with intact and complete antigens from T-ALL-derived iPSCs(T-ALL-iPSCs).We evaluated the safety and antitumor efficiency of autologous tumor-derived iPSC antigens by flow cytometry,cytokine release assay,acute toxicity experiments,long-term toxicity experiments,and other methods.Results:Our results indicate that complete tumor antigens from T-ALL-iPSCs could inhibit the growth of inoculated tumors in immunocompromised mice without causing acute and long-term toxicity.Conclusion:T-ALL-iPSC-based treatment is safe and can be used as a potential strategy for leukemia immunotherapy.

Penile cancer:Updates in systemic therapy

Penile cancer is a rare genitourinary malignancy with a greater incidence in parts of Asia,South America,and Africa.Outcomes are very poor in patients with advanced disease and in those who do not respond to frstine mutimodal therapy.Among systemic therapy options,platinum-based chemotherapy is used in the frst line;however,approximately half of patients do not benefit.Response rates to systeric therapy as subsequent line treatment are historically dismal.There is also a paucity of prognostic and predictive tools within the context of penile cancer.As such,there remains an urgent need to expand systemic treatment options for patients with advanced penile cancer.The purpose of this review is to summarize the existing evidence for standard-of-care lines of systemic treatment,examine the potential of novel lines of systemic therapy,and provide an update as to the status of these new therapies within the context of penile cancer.

Nanotherapeutics approaches to improve the efficacy of CAR-T cells in solid tumors

Adoptive cell therapy and Immune Checkpoint Blockade Inhibitors have recently revolutionized the field of oncology.However,these types of immunotherapeutic approaches have limited success in treating solid tumors.In particular,chimeric antigen receptor(CAR)-T cells efficacy is hampered by immunosuppressive signals in the tumor microenvironment(TME)and by a limited infiltration of re-infused T cells to the tumor site.The field of nanobiotechnology applied to oncology is also rapidly expanding.Nanoparticles-based delivery systems can be employed to modulate the activity of immune cells present in the TME enhancing the efficacy of CAR-T cells.Interestingly,nano-backpacks can be attached to CAR-T cells prior to re-infusion to support their homing to the tumor site and to slowly release immunopotentiators directly in the TME.Furthermore,nanovaccines can also be employed to support the in vivo expansion of CAR-T cells with consequent enhancement of their therapeutic potential.In this viewpoint,recent advancement in the field of nanobiotechnology to support CAR-T cell therapy will be discussed.The development of novel therapeutic CAR-T cells protocols together with nanotherapies is warranted in order to take full advantage of the high therapeutic potential of CAR-T cell therapy.

Personalized immunotherapy in cancer precision medicine

With the significant advances in cancer genomics using next-generation sequencing technologies,genomic and molecular profilingbased precision medicine is used as a part of routine clinical test for guiding and selecting the most appropriate treatments for individual cancer patients.Although many molecular-targeted therapies for a number of actionable genomic alterations have been developed,the clinical application of such information is still limited to a small proportion of cancer patients.In this review,we summarize the current status of personalized drug selection based on genomic and molecular profiling and highlight the challenges how we can further utilize the individual genomic information.Cancer immunotherapies,including immune checkpoint inhibitors,would be one of the potential approaches to apply the results of genomic sequencing most effectively.Highly cancer-specific antigens derived from somatic mutations,the so-called neoantigens,occurring in individual cancers have been in focus recently.Cancer immunotherapies,which target neoantigens,could lead to a precise treatment for cancer patients,despite the challenge in accurately predicting neoantigens that can induce cytotoxic T cells in individual patients.Precise prediction of neoantigens should accelerate the development of personalized immunotherapy including cancer vaccines and T-cell receptor-engineered T-cell therapy for a broader range of cancer patients.

Cellular based treatment modalities for unresectable hepatocellular carcinoma

Hepatocellular carcinoma(HCC)is the most common primary malignancy of the liver and is unfortunately associated with an overall poor prognosis and high mortality.Early and intermediate stages of HCC allow for treatment with surgical resection,ablation and even liver transplantation,however disease progression warrants conventional systemic therapy.For years treatment options were limited to molecular-targeting medications,of which sorafenib remains the standard of care.The recent development and success of immune checkpoint inhibitors has proven to be a breakthrough in the treatment of HCC,but there is an urgent need for the development of further novel therapeutic treatments that prolong overall survival and minimize recurrence.Current investigation is focused on adoptive cell therapy including chimeric antigen receptor-T cells(CAR-T cells),T cell receptor(TCR)engineered T cells,dendritic cells,natural killer cells,and tumor infiltrating lymphocyte cells,which have shown remarkable success in the treatment of hematological and solid tumor malignancies.In this review we briefly introduce readers to the currently approved systemic treatment options and present clinical and experimental evidence of HCC immunotherapeutic treatments that will hopefully one day allow for revolutionary change in the treatment modalities used for unresectable HCC.We also provide an up-to-date compilation of ongoing clinical trials investigating CAR-T cells,TCR engineered T cells,cancer vaccines and oncolytic viruses,while discussing strategies that can help overcome commonly faced challenges when utilizing cellular based treatments.

Anti-cd TCR antibody-expanded cd T cells:a better choice for the adoptive immunotherapy of lymphoid malignancies

Cell-based immunotherapy for lymphoid malignancies has gained increasing attention as patients develop resistance to conventional treatments.cd T cells,which have major histocompatibility complex(MHC)-unrestricted lytic activity,have become a promising candidate population for adoptive cell transfer therapy.We previously established a stable condition for expanding cd T cells by using anti-cd T-cell receptor(TCR)antibody.In this study,we found that adoptive transfer of the expanded cd T cells to Daudi lymphoma-bearing nude mice significantly prolonged the survival time of the mice and improved their living status.We further investigated the characteristics of these antibody-expanded cd T cells compared to the more commonly used phosphoantigen-expanded cd T cells and evaluated the feasibility of employing them in the treatment of lymphoid malignancies.Slow but sustained proliferation of human peripheral blood cd T cells was observed upon stimulation with anti-cd TCR antibody.Compared to phosphoantigen-stimulated cd T cells,the antibody-expanded cells manifested similar functional phenotypes and cytotoxic activity towards lymphoma cell lines.It is noteworthy that the anti-cd TCR antibody could expand both the Vd1 and Vd2 subsets of cd T cells.The in vitro-expanded Vd1 T cells displayed comparable tumour cell-killing activity to Vd2 T cells.Importantly,owing to higher C–C chemokine receptor 4(CCR4)and CCR8 expression,the Vd1 T cells were more prone to infiltrate CCL17-or CCL22-expressing lymphomas than the Vd2 T cells.Characterizing the peripheral blood cd T cells from lymphoma patients further confirmed that the anti-cd TCR antibody-expanded cd T cells could be a more efficacious choice for the treatment of lymphoid malignancies than phosphoantigen-expanded cd T cells.

Enhancement of T cell infiltration via tumor-targeted Th9 cell delivery improves the efficacy of antitumor immunotherapy of solid tumors

Insufficient infiltration of T cells severely compromises the antitumor efficacy of adoptive cell therapy(ACT)against solid tumors.Here,we present a facile immune cell surface engineering strategy aiming to substantially enhance the anti-tumor efficacy of Th9-mediated ACT by rapidly identifying tumor-specific binding ligands and improving the infiltration of infused cells into solid tumors.Non-genetic decoration of Th9 cells with tumor-targeting peptide screened from phage display not only allowed precise targeted ACT against highly heterogeneous solid tumors but also substantially enhanced infiltration of CD8+T cells,which led to improved antitumor outcomes.Mechanistically,infusion of Th9 cells modified with tumor-specific binding ligands facilitated the enhanced distribution of tumor-killing cells and remodeled the immunosuppressive microenvironment of solid tumors via IL-9 mediated immunomodulation.Overall,we presented a simple,cost-effective,and cell-friendly strategy to enhance the efficacy of ACT against solid tumors with the potential to complement the current ACT.

Natural killer cell-derived extracellular vesicle significantly enhanced adoptive T cell therapy against solid tumors via versatilely immunomodulatory coordination

Insufficient tumor tropism,MHC classⅠmolecules(MHC-I)defects of tumor cells,and immunosuppressive tumor microenvironment(TME)seriously imperil the efficacy of adoptive T cell therapy on solid tumors.Here,natural killer cell-derived extracellular vesicle(Nev)is used as a versatile toolkit to synergistically improve adoptive T cell therapy for solid tumors.Specifically,Nev is modified with dibenzocyclooctynes(DBCO)linked with p H-sensitive benzoic-imine bonds;meanwhile,cytotoxic T lymphocyte(CTL)is decorated with azide groups.Then CTL is armed with Nev(CTL-Nev)through the click chemistry reaction.After systematic administration,Nev obviously promotes the tumor-targeting accumulation of CTL coming from its native tumor-tropism capability.Then,the cleavage of benzoic-imine bonds in the slightly acidic TME leads to the release of Nev,which not only directly induces tumor apoptosis but also promotes the action of CTL via multiplex pathways,such as up-regulating the MHC-I expression on tumor cells,reprogramming tumor-associated macrophages from pro-tumoral M2 phenotypes to tumoricidal M1 phenotypes.The all-around coordination of Nev with CTL results in potent tumor repression.

Chimeric antigen receptor- and natural killer cell receptor-engineered innate killer cells in cancer immunotherapy

Chimeric antigen receptor(CAR)-engineered T-cell(CAR-T)therapy has demonstrated impressive therapeutic efficacy against hematological malignancies,but multiple challenges have hindered its application,particularly for the eradication of solid tumors.Innate killer cells(IKCs),particularly NK cells,NKT cells,andγδT cells,employ specific antigen-independent innate tumor recognition and cytotoxic mechanisms that simultaneously display high antitumor efficacy and prevent tumor escape caused by antigen loss or modulation.IKCs are associated with a low risk of developing GVHD,thus offering new opportunities for allogeneic“off-the-shelf”cellular therapeutic products.The unique innate features,wide tumor recognition range,and potent antitumor functions of IKCs make them potentially excellent candidates for cancer immunotherapy,particularly serving as platforms for CAR development.In this review,we first provide a brief summary of the challenges hampering CAR-T-cell therapy applications and then discuss the latest CAR-NK-cell research,covering the advantages,applications,and clinical translation of CAR-and NK-cell receptor(NKR)-engineered IKCs.Advances in synthetic biology and the development of novel genetic engineering techniques,such as gene-editing and cellular reprogramming,will enable the further optimization of IKC-based anticancer therapies.

Immunotherapy in non-small cell lung cancer:rationale,recent advances and future perspectives

Lung cancer,with non-small cell lung cancer(NSCLC)being the major type,is the second most commonmalignancy and the leading cause of cancer-related death globally.Immunotherapy,represented by immune checkpoint inhibitors(ICIs),has been one of the greatest advances in recent years for the treatment of solid tumors including NSCLC.However,not all NSCLC patients experience an effective response to immunotherapy with the established selection criteria of programmed death ligand 1(PD-L1)and tumor mutational burden(TMB).Furthermore,a considerable proportion of patients experience unconventional responses,including pseudoprogression or hyperprogressive disease(HPD),immune-related toxicities,and primary or acquired resistance during the immunotherapy process.To better understand the immune response in NSCLC and provide reference for clinical decision-making,we herein review the rationale and recent advances in using immunotherapy to treat NSCLC.Moreover,we discuss the current challenges and future strategies of this approach to improve its efficacy and safety in treating NSCLC.

The Tumor Infiltrating Lymphocytes (TILs): Did We Find the Missed Piece of the Huge Puzzle?

Tumor infiltrating lymphocytes (TILs) are used in evaluating the prognosis and determining treatment of different types of cancer with variable degrees of success. The usage of checkpoint inhibitor immunotherapy as a treatment variety for cancer and Adoptive cell therapy is associated with many complications, severe side effects and high expenses. Recently, in a limited number of metastatic GIT and breast cancers, the identification of T-cell specific against so-called tumor neo-antigens and Adoptive transfer of those lymphocytes resulted in some improvement. In 2020, Detection of a T cell receptor (TCR) in a T cell clone that recognized and killed most human cancer cell lines in vitro via the monomorphic MHC class I-related protein MR1, offers an opportunity for pan-cancer therapy Twenty three years earlier, Moist Heat was used successfully to activate a whole different and new immune response that was able to detect genetic mutation in the affected cancer cells and cured many cases of squamous and basal cell carcinomas. In this commentary review, we aimed to revise the literature for updates of TILs usage in cancer prognosis and treatment.

Personalized T-cell therapy in liver transplanted patients with hepatitis B virus related hepatocellular carcinoma

Hepatocellular carcinoma(HCC)is a deadly malignancy which typically occurs in the context of chronic liver inflammation.Chronic hepatitis B virus(HBV)infection is considered a major global cause of HCC development.At the moment,liver transplantation is the only curative modality for HBV-associated HCC.However,some patients develop HBV-HCC recurrence after liver transplantation,leaving them with very limited therapeutic options.Adoptive cell therapy with HBV-specific T cell receptor(TCR)that redirects T cells against HCC relapses has shown promising results in such HBV-HCC patients.In this mini-review,we discuss the application of this personalized T cell therapy,and highlight mRNA electroporation as an efficient tool for engineering safe and efficient TCR-redirected T cells for the treatment of liver transplant patients with HBV-HCC metastasis.

Dendritic cell-mimicking scaffolds for ex vivo T cell expansion

We propose an ex vivo T cell expansion system that mimics natural antigen-presenting cells(APCs)for adoptive cell therapy(ACT).Microfiber scaffolds coated with dendritic cell(DC)membrane replicate physicochemical properties of dendritic cells specific for T cell activation such as rapid recognition by T cells,long duration of T cell tethering,and DC-specific co-stimulatory cues.The DC membrane-coated scaffold is first surface-immobilized with T cell stimulatory ligands,anti-CD3(αCD3)and anti-CD28(αCD28)antibodies,followed by adsorption of releasable interleukin-2(IL-2).The scaffolds present both surface and soluble cues to T cells ex vivo in the same way that these cues are presented by natural APCs in vivo.We demonstrate that the DC-mimicking scaffold promotes greater polyclonal expansion of primary human T cells as compared toαCD3/αCD28-func-tionalized Dynabead.More importantly,major histocompatibility complex molecules derived from the DC membrane of the scaffold allow antigen-specific T cell expansion with target cell-specific killing ability.In addition,most of the expanded T cells(~97%)can be harvested from the scaffold by density gradient centri-fugation.Overall,the DC-mimicking scaffold offers a scalable,modular,and customizable platform for rapid expansion of highly functional T cells for ACT.

Adaptive T cell immunotherapy in cancer

Impaired tumor-specific effector T cells contribute to tumor progression and unfavorable clinical outcomes. As a compensatoryT cell-dependent cancer immunoediting strategy, adoptive T cell therapy (ACT) has achieved encouraging therapeutic results,and this strategy is now on the center stage of cancer treatment and research. ACT involves the ex vivo stimulation and expansionof tumor-infiltrating lymphocytes (TILs) with inherent tumor reactivity or T cells that have been genetically modified to expressthe cognate chimeric antigen receptor or T cell receptor (CAR/TCR), followed by the passive transfer of these cells into alymphodepleted host. Primed T cells must provide highly efficient and long-lasting immune defense against transformed cellsduring ACT. Anin-depth understanding of the basic mechanisms of these living drugs can help us improve upon currentstrategies and design better next-generation T cell-based immunotherapies. From this perspective, we provide an overview ofcurrent developments in different ACT strategies, with a focus on frontier clinical trials that offer a proof of principle. Meanwhile,insights into the determinants of ACT are discussed, which will lead to more rational, potent and widespread applicationsin the future.