Artificial antigen-presenting cells plus IL-15 and IL-21 efficiently induce melanoma-specific cytotoxic CD8^+CD28^+ T lymphocyte responses

Objective:To develop a novel artificial antigen-presenting system for efficiently inducing melanoma-specific CD8+CD28+ cytotoxic T lymphocyte（CTL） responses.Methods:Cell-sized Dynabeads? M-450 Epoxy beads coated with H-2Kb:Ig-TRP2(181111K and anti-CD28 antibody were used as artificial antigen-presenting cells（aAPCs） lo induce melanoma-specific CD8*CD28’ CTL responses with the help of IL-2I and IL-I5.Dimer staining,proliferation,ELISPOT,and cytotoxicity experiments were conducted to evaluate the frequency and activity of induced CTLs.Results:Dimer staining demonstrated that the new artificial antigen-presenting system efficiently induced melanoma TRP2-specific CD8CD28' CTLs.Proliferation and ELISPOT assays indicated that the induced CTLs rapidly proliferate and produce increased IFN- y under the slimulalion of H-2K:Ig-TRP2-aAPCs,TL-15,and IL-21.In addition,cytoloxicily experiments showed lhat induced CTLs have specific killing activity of target cells.Conclusions:The new artificial antigen-presenting system including aAPCs plus IL-21 and IL-15 can induce a large number of antigen-specific CD8+CD28+ CTLs against the melanoma.Our study provides evidence for a novel adoptive immunotherapy against tumors.

Cancer-testis Antigen OY-TES-1 Expression and Immunogenicity in Hepatocellular Carcinoma

Summary:Cancer testis(CT)antigens have received particular attention in cancer immunotherapy.OY-TES-1 is a member of CT antigens.This study was to evaluate OY-TES-1 expression and immunogenicity in hepatocelluar carcinoma(HCC).OY-TES-1 mRNA expression was detected in 56 HCC tissues and 5 normal liver tissues by reverse transcriptase PCR(RT-PCR).Of the 56 cases of HCC tissues tested,37 cases had tumor and matched adjacent non-cancer tissues and were subjected to both RT-PCR and quantitative real-time PCR.OY-TES-1 protein was subsequently observed on a panel of tissue microarrays.Sera from patients were tested for OY-TES-1 antibody by ELISA.To identify OY-TES-1 capable of inducing cellular immune response,OY-TES-1 protein was used to sensitize dentritic cells and the cytotoxicity effect was measured in vitro.The results showed that OY-TES-1 mRNA was highly expressed in 41 of the 56(73.21%)HCC tissues,whereas none in 5 normal liver tissues.OY-TES-1 mRNA was frequently expressed not only in HCC tissues(72.97%,27/37),but also in paired adjacent non-cancer tissues(64.86%,24/37).But the mean expression level of OY-TES-1 mRNA in HCC tissues was significantly higher than that in adjacent non-cancer tissues(0.76854 vs.0.09834,P=0.021).Immunohistochemistry showed that OY-TES-1 protein expression was detected in 6 of the 49 cases of HCC tissues,and absent in 9 cases of normal liver and 6 cases of cirrhosis tissues.Seropositivity was detected in 10 of the 45 HCC patients,but not detected in 17 cirrhosis patients and 76 healthy donors.The specific cytotoxic T cells elicited by OY-TES-1 could kill HLA-A2^+HCC cell line which expressed OY-TES-1.The target lysis was mainly HLA class I-dependent and could be blocked by antibodies against monomorphic HLA class I but not HLA class II molecule.In summary,OY-TES-1 expression is upregulated in HCC tissues and can be recognized by humoral and cellular responses,which suggests that OY-TES-1 is an attractive target for tumor immunotherapy in HCC.

Identification of Dysregulated microRNAs in Glioma Using RNA sequencing

Glioma is the most common malignant brain tumor in central nervous system.Despite advances in the treatment of glioma such as surgery and chemoradiotherapy,most patients are easy to relapse,resulting in adverse clinical outcomes.Hence,effective molecular=targeting treatment may be one of attractive strategies for glioma therapy.The dysregulated microRNAs(miRNAs),one of the candidates of therapeutic targets,are believed to play an important role in the progression of glioma.In this study,we aimed to examine the expression profile of miRNAs in glioma and provide a reference for glioma therapy.Firstly,expression profile of miRNAs in 5 normal brain tissues,5 low-grade glioma(LGG)tissues and 5 glioblastoma(GBM)tissues was detected by RNA sequencing(RNA-seq).Next,the target genes of differentially expressed miRNAs(DEmiRNAs)were predicted and then GO enrichment and KEGG pathway analysis performed by bioinformatics.Finally,10 miRNAs which were significantly up-or down-regulated both in GBM and LGG were validated by real-time quantitative PCR(qRT-PCR).RNA-seq results indicated a number of DEmiRNAs in glioma.There were 64 up-regulated miRNAs and 17 down-regulated miRNAs n LGG,and 181 up-regulated miRNAs and 124 down-regulated miRNAs in GBM,respectively.Bioinformatics analysis showed that the target genes of these DEmiRNAs were enriched in various biological processes and signaling pathways such as cell metabolic and developmental process.Selected DEmiRNAs were further confirmed by qRT-PCR.miRNA-10b-5p,miRNA-92b-3p and miRNA-455-5p were significantly up-regulated in both GBM and LGG;while miRNA-542-3p was significantly up-regulated in LGG;miRNA-184 and miRNA-206 were significantly down-regulated in both GBM and LGG;miRNA-766-5p and miRNA-1-3p were significantly down-regulated in GBM.The subject of our study demonstrated several dysregulated miRNAs may serve as a potential therapeutic target for gl ioma.

FMR1NB Involved in Glioma Tumorigenesis Is a Promising Target for Prognosis and Therapy

Objective:Cancer/testis antigen FMR1NB is aberrantly expressed in various types of cancer,but not in normal tissues except for testis.This study aimed to investigate the expression and functional role of FMR1NB in glioma.Methods:The expression of FMR1NB mRNA and protein was determined using RT-PCR and immunohistochemistry,respectively,in glioma specimens from 83 patients at follow-up.The effects of siRNA-mediated FMR1NB silencing on malignant biological behaviors were evaluated in glioma cell lines Al 72 and U251.Results:FMR1NB mRNA and protein expression was detected in 58.8%(77/131)and 46.34%(57/123)of glioma tissues,respectively.FMR1NB protein was positively correlated with World Health Organization grade and found to be an independent prognostic marker for poor outcome.Knockdown of FMR1NB induced apoptosis and suppressed proliferation,adhesion,migration,and invasion by modulating the expression of cyclin A,CDK2,caspase-3,E-cadherin,and N-cadherin in A172 and U251 cells.Conclusion:Our findings suggest that FMR1NB contributes to the tumorigenesis of glioma cells and may represent a potential prognostic biomarker and an attractive therapeutic target in glioma.

High Expression of Fibronectin 1 Predicts a Poor Prognosis in Glioblastoma

Objective Glioblastoma multiforme(GBM),the most malignant intracranial neoplasm,is associated with a high mortality and recurrence rate due to the aggressive nature and heterogeneity of the tumor.Some of the molecular markers involved in the tumorigenesis of GBM are essential in prognosis,diagnosis,and treatment.Due to the limitations of therapeutic effects,this study aims to explore novel biomarkers with prognostic value and to provide new insights into therapeutic targets.Methods The expression profile of mRNAs in GBM was detected by RNA-sequencing,and differentially expressed genes were identified by integrating the data from RNA-seq results and the GEPIA2 database.Of the total 40 hub genes,FN1,P4HB,and PPIB showed prognostic significance based on both GEPIA2 and CGGA databases.The validation of FN1,P4HB,and PPIB expression by qPCR and correlation analysis with clinicopathological features were performed in 41 GBM tissues from our institution.Results Kaplan-Meier analysis revealed that FN1 and P4HB expressions levels were related to the overall survival(OS)of GBM patients(P<0.05).Multivariate analysis showed that FN1 overexpression(HR=9.199,P=0.002)was an independent and unfavorable prognostic factor for GBM patients.The median survival time was 8.5 months and 21 months for high and low expressions of FN1,respectively.Conclusion It was suggested that FN1 could be an ideal target for prognosis and a potential therapeutic target in GBM.

Microrna-1224-5p Is a Potential Prognostic and Therapeutic Biomarker in Glioblastoma:Integrating Bioinformatics and Clinical Analyses

Objective Glioblastoma(GBM)is the most common,invasive,and malignant primary brain tumor with a poor prognosis and high recurrence rate.It’s known that some microRNAs(miRNAs)which are associated with tumorigenesis and progression can be considered as prognostic and therapeutic targets in tumors including GBM.This study aims to highlight the potential role of the core miRNAs in GBM and their potential use as a prognostic and therapeutic biomarker.Methods Differentially expressed miRNAs(DEmiRNAs)were identified in GBM by integrating miRNA-sequencing results and a GBM microarray dataset from the Gene Expression Omnibus(GEO)database through bioinformatics tools.The dysregulated miRNAs were identified by survival analysis through Chinese Glioma Genome Atlas(CGGA).Target genes of the dysregulated miRNAs were predicted on MiRWalk and miRTarBase database.TAM2.0 database,Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathways analysis were used to analyze the function of the dysregulated miRNAs.Subsequently,protein-protein interaction(PPI)network analysis was used to identify the top 20 hub targets of the up-regulated and down-regulated miRNAs,respectively.Then,core miRNAs in GBM were identified by constructing dysregulated miRNA-differentially expressed hub gene networks.Validation of the core miRNAs expression was detected in 41 GBM tissues compared to 8 normal brain tissues.Furthermore,the potential biomarkers were identified by clinical correlation analysis and survival analysis.Results Totally,68 intersecting DEmiRNAs were identified,40 of which were upregulated and the other 28 miRNAs were downregulated.Two upregulated and 4 downregulated miRNAs showed prognostic significance.Most differentially expressed hub genes were regulated by the miR-28-5p and miR-1224-5p,which were respectively upregulated and downregulated in GBM.The correlation between miR-1224-5p level and recurrence was statistically significant(P=0.011).Survival analysis showed that high miR-28-5p level and high miR-1224-5p level were both associated with better prognosis.Moreover,high miR-1224-5p level was an independent prognosis factor for GBM patients according to the cox regression analysis.Conclusion MiRNA-1224-5p could be a potential target for the prognosis and treatment in GBM.