ABSTRACT
The powdery mildew caused by Eeysiphe heraclei is a serious concern in Heracleum moellendorffii Hance. Therefore, exploring the mechanisms underlying sugar efflux from host cells to the fungus during the plant-fungus interaction showed great significance. The study successfully cloned HmSWEET8 and HmSTP1 genes based on RNA-seq technology. The complementation assays in yeast EBY.VW4000 found HmSWEET8 and HmSTP1 transporting hexose. Over-expressing or silencing HmSWEET8 in H. moellendorffii leaves increased or decreased powdery mildew susceptibility by changing glucose concentration in infective sites. Meanwhile, over-expressing HmSTP1 in H. moellendorffii leaves also increased powdery mildew susceptibility by elevating the glucose content of infective areas. Additionally, HmSTP1 expression was up-regulated obviously in HmSWEET8 over-expressed plants and inhibited significantly in HmSWEET8 silenced plants. Co-expressing HmSWEET8 and HmSTP1 genes significantly increased powdery mildew susceptibility compared with over-expressed HmSWEET8 or HmSTP1 plants alone. The results demonstrated that HmSTP1 may assist with HmSWEET8 to promote E. heraclei infection. Consequently, the infection caused by E. heraclei resulted in the activation of HmSWEET8, leading to an increased transfer of glucose to the apoplasmic spaces at the sites of infection, then, HmSTP1 facilitated the transport of glucose into host cells, promoting powdery mildew infection.
ABSTRACT
The endothelial barrier plays a critical role in immune defense against bacterial infection. Efficient interactions between neutrophils and endothelial cells facilitate the activation of both cell types. However, neutrophil activation can have dual effects, promoting bacterial clearance on one hand while triggering inflammation on the other. In this review, we provide a detailed overview of the cellular defense progression when neutrophils encounter bacteria, focusing specifically on neutrophil-endothelial interactions and endothelial activation or dysfunction. By elucidating the underlying mechanisms of inflammatory pathways, potential therapeutic targets for inflammation caused by endothelial dysfunction may be identified. Overall, our comprehensive understanding of neutrophil-endothelial interactions in modulating innate immunity provides deeper insights into therapeutic strategies for infectious diseases and further promotes the development of antibacterial and anti-inflammatory drugs.
ABSTRACT
Epigenetic modification shapes differentiation trajectory and regulates the exhaustion state of chimeric antigen receptor T (CAR-T) cells. Limited efficacy induced by terminal exhaustion closely ties with intrinsic transcriptional regulation. However, the comprehensive regulatory mechanisms remain largely elusive. Here, we identify class I histone deacetylase inhibitors (HDACi) as boosters of CAR-T cell function by high-throughput screening of chromatin-modifying drugs, in which M344 and chidamide enhance memory maintenance and resistance to exhaustion of CAR-T cells that induce sustained antitumor efficacy both in vitro and in vivo. Mechanistically, HDACi decrease HDAC1 expression and enhance H3K27ac activity. Multi-omics analyses from RNA-seq, ATAC-seq, and H3K27ac CUT&Tag-seq show that HDACi upregulate expression of TCF4, LEF1, and CTNNB1, which subsequently activate the canonical Wnt/ß-catenin pathway. Collectively, our findings elucidate the functional roles of class I HDACi in enhancing CAR-T cell function, which provides the basis and therapeutic targets for synergic combination of CAR-T cell therapy and HDACi treatment.
Subject(s)
Aminopyridines , Histone Deacetylase Inhibitors , Wnt Signaling Pathway , Histone Deacetylase Inhibitors/pharmacology , Wnt Signaling Pathway/drug effects , Animals , Humans , Mice , Benzamides/pharmacology , Cell Line, Tumor , Immunotherapy, Adoptive/methods , Receptors, Chimeric Antigen/metabolism , T-Lymphocytes/drug effects , T-Lymphocytes/metabolism , T-Lymphocytes/immunology , Histone Deacetylase 1/metabolismABSTRACT
Due to their unique location, airway tumors have a significant impact on patient quality of life and survival. Current research has focused extensively on malignant airway tumors; however, benign airway tumors, especially rare ones, are less understood due to their low incidence. These tumors are often misdiagnosed and mistreated due to diagnostic challenges. Therefore, there is still a lack of consensus on the treatment of some rare benign airway tumors. Our center summarizes the diagnosis and treatment of four rare cases of benign airway stenosis in recent years, highlighting the bronchoscopic manifestations and therapeutic approaches to improve the understanding of these diseases.
ABSTRACT
Chimeric antigen receptor (CAR)-T therapy has shown superior efficacy against hematopoietic malignancies. However, many patients failed to achieve sustainable tumor control partially due to CAR-T cell exhaustion and limited persistence. In this study, by performing single-cell multi-omics data analysis on patient-derived CAR-T cells, we identify CD38 as a potential hallmark of exhausted CAR-T cells, which is positively correlated with exhaustion-related transcription factors and further confirmed with in vitro exhaustion models. Moreover, inhibiting CD38 activity reverses tonic signaling- or tumor antigen-induced exhaustion independent of single-chain variable fragment design or costimulatory domain, resulting in improved CAR-T cell cytotoxicity and antitumor response. Mechanistically, CD38 inhibition synergizes the downregulation of CD38-cADPR -Ca2+ signaling and activation of the CD38-NAD+-SIRT1 axis to suppress glycolysis. Collectively, our findings shed light on the role of CD38 in CAR-T cell exhaustion and suggest potential clinical applications of CD38 inhibition in enhancing the efficacy and persistence of CAR-T cell therapy.
Subject(s)
Neoplasms , Single-Chain Antibodies , Humans , T-Lymphocytes , Immunotherapy, Adoptive/methods , Antigens, Neoplasm/metabolismABSTRACT
OBJECTIVE: To investigate the effect of metformin and arsenic trioxide on KG1a cells proliferation of acute myeloid leukemia and its possible mechanism. METHODS: CCK-8 method was used to detect the killing effect of metformin, arsenic trioxide and combined application on KG1a cells. Annexin V-FITC/PI Dual Stain Flow Cytometry was used to detect the effect of combined application on apoptosis of KG1a cells. Western blot was used to detect the expression of intracellular apoptosis-,autophagy-related protein. RESULTS: Metformin and arsenic trioxide alone or in combination could inhibit the proliferation of KG1a cells and induce apoptosis of KG1a cells, and the proliferation inhibition rate and apoptosis rate in the combined drug group were higher than those in the drug group alone(P <0.05). The combination of drugs induced upregulation of Caspase 8 protein and P62 protein expression and was higher than that in the drug group alone(P <0.05). CONCLUSION: Metformin can synergize with arsenic trioxide to kill KG1a cells, and its mechanism of action may be related to inducing apoptosis and enhancing autophagy.
Subject(s)
Arsenicals , Metformin , Humans , Arsenic Trioxide/pharmacology , Metformin/pharmacology , Oxides/pharmacology , Arsenicals/pharmacology , Cell ProliferationABSTRACT
The efficacy of chimeric antigen receptor (CAR) T cell therapy is hampered by relapse in hematologic malignancies and by hyporesponsiveness in solid tumors. Long-lived memory CAR T cells are critical for improving tumor clearance and long-term protection. However, during rapid ex vivo expansion or in vivo tumor eradication, metabolic shifts and inhibitory signals lead to terminal differentiation and exhaustion of CAR T cells. Through a mitochondria-related compound screening, we find that the FDA-approved isocitrate dehydrogenase 2 (IDH2) inhibitor enasidenib enhances memory CAR T cell formation and sustains anti-leukemic cytotoxicity in vivo. Mechanistically, IDH2 impedes metabolic fitness of CAR T cells by restraining glucose utilization via the pentose phosphate pathway, which alleviates oxidative stress, particularly in nutrient-restricted conditions. In addition, IDH2 limits cytosolic acetyl-CoA levels to prevent histone acetylation that promotes memory cell formation. In combination with pharmacological IDH2 inhibition, CAR T cell therapy is demonstrated to have superior efficacy in a pre-clinical model.
Subject(s)
Antioxidants , Neoplasms , Humans , Antioxidants/pharmacology , Antioxidants/metabolism , Isocitrate Dehydrogenase , Histones/metabolism , Acetylation , T-Lymphocytes , Neoplasms/metabolism , Mitochondria/metabolismABSTRACT
To obtain a high-performance gas sensor, it is essential to ingeniously design sensing materials containing the features of high catalytic performance, abundant oxygen vacancies, and splendid grain dispersibility through a simple method. Inspired by the fact that ZIF-8 contains semiconductor metal atoms, well-arranged ZnO nanoparticle (NP)-in situ assembled one-dimensional nanofibers (NFs) are obtained by one-step electrospinning. By incorporating Pt NPs into the cavity of ZIF-8 NPs, well-dispersed Pt@ZnO NPs driven by Pt@ZIF-8 composites are obtained after annealing. The well-arranged Pt@ZnO NP-assembled NFs not only exhibit abundant oxygen vacancies but also avoid the self-aggregation of ZnO and Pt NPs. Meanwhile, the small Pt NPs could improve the catalytic effect in return. Therefore, the gas sensor fabricated based on the above materials exhibits an acetone sensitivity of 6.1 at 370 °C, compared with pristine ZnO NFs (1.6, 5 ppm). Moreover, the well-arranged Pt@ZnO NP-assembled NFs show exceptional sensitivity to acetone with a 70.2 ppb-level detection limit in theory. The synergistic advantages of the designed sensing material open up new possibilities for non-invasive disease diagnosis.
ABSTRACT
Background: Stress urinary incontinence (SUI) is a common condition that can severely affect women's life quality. Electroacupuncture (EA) has been proved to be an optional treatment for SUI, but the tolerance of EA becomes a factor affecting efficiency, which should not be ignored and needs to be solved urgently. The purpose of this study is to find out whether the use of alternating acupoints combination can solve this problem or not and provide an optimization of EA treatment for female SUI. Methods: This multi-center randomized controlled trial will enroll 360 patients with SUI. They will be randomly assigned to one of the three groups-sacral acupoints group (sacral group), abdominal acupoints group (abdominal group), or alternating acupoints group (alternating group)-at a 1:1:1 ratio. The patients will receive 18 sessions of EA treatment and will be followed up for 48 weeks after the treatment. The primary outcome measure of the study is the change of urine leakage at week 6. The secondary outcomes include the incontinence episode frequency (IEF), International Consultation on Incontinence Questionnaire-Short Form (ICIQ-SF), severity of SUI, patient self-evaluation of therapeutic effects, weekly usage of urine pads, ultrasonography of pelvic floor, specialty therapies for SUI, evaluation of discomfort during EA treatment, patient acceptability evaluation and adverse events related to intervention. Discussion: This trial is specifically designed to offer an optimized EA treatment for female SUI, aiming to enhance their quality of life.Clinical trial registration: ClinicalTrials.gov, identifier ID:NCT05635669.
ABSTRACT
Proteins are considered major effectors of sperm function. However, the proteins expressed in pigeon sperm have not been explored. Here, we collected semen from meat and racing pigeons using the electroejaculation method and identified proteins in pigeon sperm using the proteomics approach. A total of 1,641 proteins were identified in the sperm of domesticated pigeons. Of which, 1,541 proteins were reliably quantified, and gene ontology (GO) and associated bioinformatics analyses indicated that annotated proteins were linked to the oxidation-reduction process, integral component membrane, and protein binding, etc. Among quantified proteins, 1,515 and 1,507 proteins were respectively presented in White King pigeons and racing pigeons, and 1,481 proteins were shared between these 2 types of pigeons, including axonemal dynein, solute carrier, cilia- and flagella-associated protein, outer dense fiber protein, etc. Proteins in our constructed protein-protein interaction (PPI) network are involved in oxidative phosphorylation, sperm axoneme assembly, cilium-dependent cell motility, axonemal dynein complex assembly, flagellated sperm motility, etc. In conclusion, this study characterized the sperm proteome of pigeons and provided a foundation for the subsequent research screening markers for fertility evaluation of pigeons.
ABSTRACT
Spermidine is a class of biologically active organic small molecules that play an important role in maintaining intestinal homeostasis. The specific objective of this study was to explore the effects of spermidine on intestinal morphology, metabolites, and microbial diversity in mice. We showed that 0.3 mmol/L of spermidine significantly promoted the growth of ileal villi (p < 0.05), and 3.0 mmol/L of spermidine significantly increased the body weight of mice and promoted the growth of jejunum villi (p < 0.05). The 16S rDNA sequencing results indicated that 3.0 mmol/L of spermidine affected the balance of the intestinal flora by increasing the abundance of intestinal Lactic acid bacteria and reducing the abundance of harmful bacteria (Turicibacter and Alistipes). Additionally, spermidine affects the levels of microbial metabolites such as succinic acid and Pantetheine. In summary, spermidine affects intestinal morphology and regulates intestinal flora and metabolites, and this study has provided a new understanding of spermidine's effects on the intestinal tract.
Subject(s)
Gastrointestinal Microbiome , Spermidine , Spermidine/pharmacology , Intestinal Mucosa/metabolism , Ileum , Jejunum , Gastrointestinal Microbiome/physiologyABSTRACT
Drug resistance is a major obstacle to the successful treatment of cancer. The role of the miR-106b-25 cluster in drug resistance of haematologic malignancies has not yet been elucidated. Here, we show that the miR-106b-25 cluster mediates resistance to therapeutic agents with structural and mechanistic dissimilarity in vitro and in vivo. RNA sequencing data revealed that overexpression of the miR-106b-25 cluster or its individual miRNAs resulted in downregulation of multiple key regulators of apoptotic pathways. Luciferase reporter assay identified TP73 as a direct target of miR-93 and miR-106b, BAK1 as a direct target of miR-25 and CASP7 as a direct target of all three miRNAs. We also showed that inhibitors of the miR-106b-25 cluster and BCL-2 exert synergistic effects on apoptosis induction in primary myeloid leukaemic cells. Thus, the members of the miR-106b-25 cluster may jointly contribute to myeloid leukaemia drug resistance by inactivating multiple apoptotic genes. Targeting this cluster could be a promising combination strategy in patients resistant to therapeutic agents that induce apoptosis.
Subject(s)
Leukemia, Myeloid , MicroRNAs , Neoplasms , Humans , MicroRNAs/metabolism , Apoptosis/genetics , Leukemia, Myeloid/drug therapy , Leukemia, Myeloid/genetics , Drug Resistance , Cell Line, Tumor , Cell ProliferationABSTRACT
Chimeric Antigen-Receptor (CAR) T-cell therapies have shown dramatic efficacy in treating relapsed and refractory cancers, especially B cell malignancies. However, these innovative therapies cause adverse toxicities that limit the broad application in clinical settings. Hematologic cytopenias, one frequently reported adverse event following CAR T cell treatment, are manifested as a disorder of hematopoiesis with decreased number of mature blood cells and subdivided into anemia, thrombocytopenia, leukopenia, and neutropenia, which increase the risk of infections, fatigue, bleeding, fever, and even fatality. Herein, we initially summarized the symptoms, etiology, risk factors and management of cytopenias. Further, we elaborated the cellular and molecular mechanisms underlying the initiation and progression of cytopenias following CAR T cell therapy based on previous studies about acquired cytopenias. Overall, this review will facilitate our understanding of the etiology of cytopenias and shed lights into developing new therapies against CAR T cell-induced cytopenias.
Subject(s)
Anemia , Hematologic Neoplasms , Neoplasms , Neutropenia , Thrombocytopenia , Hematologic Neoplasms/therapy , Humans , Immunotherapy, Adoptive/adverse effects , Neoplasms/etiology , T-Lymphocytes , Thrombocytopenia/etiologyABSTRACT
Chimeric antigen receptor T-cell (CAR-T) treatment has revolutionized the landscape of cancer therapy with significant efficacy on hematologic malignancy, especially in relapsed and refractory B cell malignancies. However, unexpected serious toxicities such as cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS) still hamper its broad application. Clinical trials using CAR-T cells targeting specific antigens on tumor cell surface have provided valuable information about the characteristics of ICANS. With unclear mechanism of ICANS after CAR-T treatment, unremitting efforts have been devoted to further exploration. Clinical findings from patients with ICANS strongly indicated existence of overactivated peripheral immune response followed by endothelial activation-induced blood-brain barrier (BBB) dysfunction, which triggers subsequent central nervous system (CNS) inflammation and neurotoxicity. Several animal models have been built but failed to fully replicate the whole spectrum of ICANS in human. Hopefully, novel and powerful technologies like single-cell analysis may help decipher the precise cellular response within CNS from a different perspective when ICANS happens. Moreover, multidisciplinary cooperation among the subjects of immunology, hematology, and neurology will facilitate better understanding about the complex immune interaction between the peripheral, protective barriers, and CNS in ICANS. This review elaborates recent findings about ICANS after CAR-T treatment from bed to bench, and discusses the potential cellular and molecular mechanisms that may promote effective management in the future. This article is categorized under: Cancer > Biomedical Engineering Immune System Diseases > Molecular and Cellular Physiology Neurological Diseases > Molecular and Cellular Physiology.
Subject(s)
Neoplasms , Neurotoxicity Syndromes , Receptors, Chimeric Antigen , Animals , Humans , Receptors, Chimeric Antigen/metabolism , Immunotherapy, Adoptive/adverse effects , Receptors, Antigen, T-Cell/metabolism , Neurotoxicity Syndromes/etiology , ConfusionABSTRACT
Myeloid-derived suppressor cells (MDSCs), a population of myeloid lineage cells with immunosuppressive capacity, can mitigate acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). We previously found that the immunosuppressive function of polymorphonuclear population (PMN-MDSCs) was impaired in aGVHD milieu. The aim of this study was to explore the intrinsic mechanism regulating the fate and function of donor-derived PMN-MDSCs during allo-HSCT. We firstly found that mitochondrial permeability transition pore (MPTP) opened in the PMN-MDSCs in response to the intense inflammatory environment of aGVHD, which induced mitochondrial damage, oxidative stress, and apoptosis of PMN-MDSCs. Inhibiting MPTP opening by a traditional immunosuppressant, cyclosporine A (CsA), could restore the immunosuppressive function and viability of PMN-MDSCs in vitro and in vivo, which reveals a new mechanism of CsA application.
Subject(s)
Graft vs Host Disease , Myeloid-Derived Suppressor Cells , Humans , Cyclosporine/pharmacology , Graft vs Host Disease/drug therapy , Immunosuppressive Agents/pharmacology , Mitochondrial Permeability Transition PoreABSTRACT
Short persistence and early exhaustion of T cells are major limits to the efficacy and broad application of immunotherapy. Exhausted T and chimeric antigen receptor (CAR)-T cells upregulate expression of genes associated with terminated T cell differentiation, aerobic glycolysis and apoptosis. Among cell exhaustion characteristics, impaired mitochondrial function and dynamics are considered hallmarks. Here, we review the mitochondrial characteristics of exhausted T cells and particularly discuss different aspects of mitochondrial metabolism and plasticity. Furthermore, we propose a novel strategy of rewiring mitochondrial metabolism to emancipate T cells from exhaustion and of targeting mitochondrial plasticity to boost CAR-T cell therapy efficacy.
Subject(s)
Immunotherapy, Adoptive , T-Lymphocytes , Humans , Immunotherapy , Lymphocyte Activation , Mitochondria/metabolismABSTRACT
Immunotherapy using chimeric antigen receptor (CAR) T cells is a rapidly emerging modality that engineers T cells to redirect tumor-specific cytotoxicity. CAR T cells have been well characterized for their efficacy against B cell malignancies, and rigorously studied in other types of tumors. Preclinical evaluation of CAR T cell function, including direct tumor killing, cytokine production, and memory responses, is crucial to the development and optimization of CAR T cell therapies. Such comprehensive examinations are usually performed in different types of models. Model establishment should focus on key challenges in the clinical setting and the capability to generate reliable data to indicate CAR T cell therapeutic potency in the clinic. Further, modeling the interaction between CAR T cells and tumor microenvironment provides additional insight for the future endeavors to enhance efficacy, especially against solid tumors. This review will summarize both in vitro and in vivo models for CAR T cell functional evaluation, including how they have evolved with the needs of CAR T cell research, the information they can provide for preclinical assessment of CAR T cell products, and recent technology advances to test CAR T cells in more clinically relevant models.
Subject(s)
Neoplasms , Receptors, Chimeric Antigen , Humans , Immunotherapy, Adoptive , Neoplasms/pathology , Neoplasms/therapy , Receptors, Antigen, T-Cell/metabolism , T-Lymphocytes , Tumor MicroenvironmentABSTRACT
The extensive application of biochar has drawn more attentions on its potential risk to aquatic organisms. However, the influence of environmental factors (i.e. pH, HA, SDBS and aging time) after they discharged into environment on their toxicity have not been clarified. Here, we synthesized biochar with local pine needles via pyrolysis, and then aged in different media. Followed, the toxicity of pristine and aged biochar was checked with Scenedesmus obliquus (S. obliquus). Our investigation showed that the toxicity of biochar was mitigated when aged in different pH levels or SDBS, while it was opposite in the presence of HA. The increment of pH decreased the toxicity of both the pristine and the aged biochar, while the presence of HA did same impact on the pristine biochar. The presence of SDBS decreased the toxicity of pristine biochar but increased that of aged biochar. Meanwhile, we showed these environmental factors (pH, HA, SDBS and aging time) influenced the biochar toxicity may be due to the adjustment of the aggregation and adhesion of biochar on cell surfaces or the intracellular oxidative stress. Further, the PFRs contained in biochar did influence the toxicity, along with the physicochemical properties of biochar (i.e. carbon structure, functional group or surface charge). Our results aimed to reflect the toxicity profile of biochar in the natural aquatic environment, without misunderstanding of potential ecological risk of biochar in the future application.
Subject(s)
Charcoal , Pyrolysis , Aquatic OrganismsABSTRACT
Chimeric antigen receptor (CAR) T cells are potent agents for recognizing and eliminating tumors, and have achieved remarkable success in the treatment of patients with refractory leukemia and lymphoma. However, dysfunction of T cells, including exhaustion, is an inevitable obstacle for persistent curative effects. Here, the authors initially found that calcium signaling is hyperactivated via sustained tonic signaling in CAR-T cells. Next, it is revealed that the store-operated calcium entry (SOCE) inhibitor BTP-2, but not the calcium chelator BAPTA-AM, markedly diminishes CAR-T cell exhaustion and terminal differentiation of CAR-T cells in both tonic signaling and tumor antigen exposure models. Furthermore, BTP-2 pretreated CAR-T cells show improved antitumor potency and prolonged survival in vivo. Mechanistically, transcriptome and metabolite analyses reveal that treatment with BTP-2 significantly downregulate SOCE-calcineurin-nuclear factor of activated T-cells (NFAT) and glycolysis pathways. Together, the results indicate that modulating the SOCE-calcineurin-NFAT pathway in CAR-T cells renders them resistant to exhaustion, thereby yielding CAR products with enhanced antitumor potency.
Subject(s)
Calcineurin , Leukemia , Calcineurin/metabolism , Calcineurin/pharmacology , Calcium Signaling , Glycolysis , Humans , Leukemia/metabolism , Leukemia/therapy , T-Lymphocytes/metabolismABSTRACT
Acute myeloid leukemia (AML) is a malignant tumor of the hematopoietic system, and leukemia stem cells are responsible for AML chemoresistance and relapse. KG-1a cell is considered a leukemia stem cell-enriched cell line, which is resistant to chemotherapy. Arsenic trioxide (ATO) is effective against acute promyelocytic leukemia as a first-line treatment agent, even as remission induction of relapsed cases. ATO has a cytotoxic effect on KG-1a cells, but the mechanism remains unclear. Our results demonstrated that ATO can inhibit cell proliferation, induce apoptosis, and arrest KG-1a cells in the G2/M phase. Using transcriptome analysis, we investigated the candidate target genes regulated by ATO in KG-1a cells. The expression profile analysis showed that the ATO had significantly changed gene expression related to proliferation, apoptosis, and cell cycle. Moreover, MYC, PCNA, and MCM7 were identified as crucial hub genes through protein-protein interaction network analysis; meanwhile, the expressions of them in both RNA and protein levels are down-regulated as confirmed by quantitative polymerase chain reaction and Western blot. Thus, our study suggests that ATO not only inhibits the expression of MYC, PCNA, and MCM7 but also leads to cell cycle arrest and apoptosis in KG-1a cells. Overall, this study provided reliable clues for improving the ATO efficacy in AML.