Prediction of Tumor Microenvironment Characteristics and Treatment Response in Lung Squamous Cell Carcinoma by Pseudogene OR7E47P-related Immune Genes

Objective Pseudogenes are initially regarded as nonfunctional genomic sequences,but some pseudogenes regulate tumor initiation and progression by interacting with other genes to modulate their transcriptional activities.Olfactory receptor family 7 subfamily E member 47 pseudogene(OR7E47P)is expressed broadly in lung tissues and has been identified as a positive regulator in the tumor microenvironment(TME)of lung adenocarcinoma(LUAD).This study aimed to elucidate the correlation between OR7E47P and tumor immunity in lung squamous cell carcinoma(LUSC).Methods Clinical and molecular information from The Cancer Genome Atlas(TCGA)LUSC cohort was used to identify OR7E47P-related immune genes(ORIGs)by weighted gene correlation network analysis(WGCNA).Based on the ORIGs,2 OR7E47P clusters were identified using non-negative matrix factorization(NMF)clustering,and the stability of the clustering was tested by an extreme gradient boosting classifier(XGBoost).LASSO-Cox and stepwise regressions were applied to further select prognostic ORIGs and to construct a predictive model(ORPScore)for immunotherapy.The Botling cohorts and 8 immunotherapy cohorts(the Samstein,Braun,Jung,Gide,IMvigor210,Lauss,Van Allen,and Cho cohorts)were included as independent validation cohorts.Results OR7E47P expression was positively correlated with immune cell infiltration and enrichment of immune-related pathways in LUSC.A total of 57 ORIGs were identified to classify the patients into 2 OR7E47P clusters(Cluster 1 and Cluster 2)with distinct immune,mutation,and stromal programs.Compared to Cluster 1,Cluster 2 had more infiltration by immune and stromal cells,lower mutation rates of driver genes,and higher expression of immune-related proteins.The clustering performed well in the internal and 5 external validation cohorts.Based on the 7 ORIGs(HOPX,STX2,WFS,DUSP22,SLFN13,GGCT,and CCSER2),the ORPScore was constructed to predict the prognosis and the treatment response.In addition,the ORPScore was a better prognostic factor and correlated positively with the immunotherapeutic response in cancer patients.The area under the curve values ranged from 0.584 to 0.805 in the 6 independent immunotherapy cohorts.Conclusion Our study suggests a significant correlation between OR7E47P and TME modulation in LUSC.ORIGs can be applied to molecularly stratify patients,and the ORPScore may serve as a biomarker for clinical decision-making regarding individualized prognostication and immunotherapy.

ANTITUMOR IMMUNITY AND VACCINE EFFECT INDUCED BY IL-12 SYNERGIZES B7-1 GENE TRANSFECTED CELLS

Objective: To study the synergic effects of IL-12 and B7-1 transfectant on antitumor immunity in vivo. Methods: The retrovirus vector encoding mIL-12 and mB7-1 gene was tranfected into EL-4 thymic lymphoma cells respectively.The cells were used as tumor vaccine and the therapeutic effect was observed. Results: In contrast to the miceimmunized with EL-4/Wt or EL-4/Neo groups, thetumorigenicity of EL-4/IL-12 transfectant was decreased(P<0.001). The EL-4/IL-12 and EL-4/B7-1 cells irradiatedwith 60Co showed significant systematic protective effectsagainst the rechallenge of EL-4/Wt. 60Co irradiatedEL-4/IL-12 cells delayed the occurrence of tumor andprolonged the survival period of tumor bearing mice.Combination of the vaccines of EL-4/IL-12 and EL-4/B7-1 resulted in the enhanced therapeutic effect compared witheach single transfectant group (P<0.001). Conclusion: The results showed that IL-12 transduced cells could enhancethe antitumor immunity of host as cancer vaccine.Combination of the EL-4/IL-12 and EL-4/B7-1 transfectant could improve immunity of host and is a prospect cancervaccine.

Preliminary Study on the Cooperation of IL-6 and mB7-1 in the Induction of Effective Antitumor Immunity In Vitro

To find a new way for gene therapy against tumors with weak immunogenicity, the effect of mB7 1 costimulation alone, or combined with IL 6, in inducing antitumor immunity in vitro was investigated. It was found that mB7 1 cDNA transfected B16 cells (B16 mB7 1) induced the expansion of effector lymphocytes and the generation of specific lytic activity more effectively than wild type B16 melanoma cells (B16 wt) or mock transfected B16 cells (B16 neo) did. ( P <0.01), IL 6 could effectively stimulate lymphocytes proliferation, but failed to enhance its cytotoxicity, while the combination of mB7 1 and IL 6 increased both lymphocyte proliferative response and T cell mediated cytotoxicity more significantly than B7 1 or IL 6 did alone ( P <0.01) . It was inferred that the costimulatory molecule B7 1 is required for the activation and proliferation of T lymphocytes; the expression of mB7 1 in tumor cells could increase their immunogenicity and induce effective antitumor immune response, and the combination of B7 1 and IL 6 could induce more effective antitumor immunity, indicating that cooperation of IL 6 and mB7 1 plays a role in T lymphocyte activation.

基于TCGA数据库分析乳腺癌组织RBP7 mRNA表达与肿瘤免疫细胞浸润及预后的相关性

目的通过生物信息学的方法探讨视黄醇结合蛋白7(retinol binding protein 7,RBP7)在乳腺癌中的作用。方法使用R语言基于癌症基因组图谱(the cancer genome atlas,TCGA)数据库和人类蛋白质图谱(the human protein atlas,HPA)数据库探索基因RBP7在乳腺癌组织中的差异表达。通过Kaplan-Meier生存分析和受试者工作特征(receiver operating characteristic,ROC)曲线,评估RBP7与乳腺癌临床数据的关系。基于TCGA数据库分析RBP7高低表达分组与不同肿瘤浸润免疫细胞(tumor-infiltrating immune cells,TIICs)的相关性。基因组富集分析(gene set enrichment analysis,GSEA)评估RBP7在与表型相关度排序的基因表中的分布趋势。结果与癌旁组织相比,乳腺癌中RBP7 mRNA表达水平下调,该分子表达在细胞核中。ROC曲线分析显示RBP7诊断乳腺癌的曲线下面积(area under curve,AUC)是0.943(95%CI:0.926~0.960),RBP7的最佳截断值是6.29,敏感度和特异度分别为82.32%,93.69%。Kaplan-Meier生存分析显示RBP7低表达与乳腺癌患者的总生存率相关(HR=0.68,95%CI:0.49~0.93,P=0.017),RBP7是乳腺癌发生的独立危险因素。Spearman相关性揭示RBP7与乳腺癌中pDC细胞和NK细胞呈正相关(r=0.290,0.253,均P<0.05),与Th2细胞呈负相关(r=-0.217,P<0.05)。GSEA表明RBP7富集在脂肪生成、核糖体、肽配体结合受体、钙信号途径等通路中(均P<0.001)。结论RBP7影响乳腺癌的发生发展,可能成为乳腺癌潜在生物标志物和治疗靶点。

Double-edge Role of B Cells in Tumor Immunity:Potential Molecular Mechanism

B cells are a heterogeneous population,which have distinct functions of antigen presentation,activating T cells,and secreting antibodies,cytokines as well as protease.It is supposed that the balance among these B cells subpopulation(resting B cells,activated B cells,Bregs,and other differentiated B cells)will determine the ultimate role of B cells in tumor immunity.There has been increasing evidence supporting opposite roles of B cells in tumor immunity,though there are no general acceptable phenotypes for them.Recent years,a new designated subset of B cells identified as Bregs has emerged from immunosuppressive and/or regulatory functions in tumor immune responses.Therefore,transferring activated B cells would be possible to become a promising strategy against tumor via conquering the immunosuppressive status of B cells in future.Understanding the potential mechanism of double-edge role of B cells will help researchers utilize activated B cells to improve their anti-tumor response.Moreover,the molecular pathways related to B cell differentiation are involved in its tumor-promoting effect,such as NF-κB,STAT3,BTK.So,we review the molecular and signaling pathway mechanisms of B cells involved in both tumor-promoting and tumor-suppressive immunity,in order to help researchers optimize B cells to fight cancer better.

Costimulatory Molecule B7-H1 on the Immune Escape of Bladder Cancer and Its Clinical Significance

B7-H1, a recently described member of the B7 family of costimulatory molecules, is thought to be involved in tumor immune escape by inducing T-cell apoptosis. In order to investigate the relationship between B7-H1 and immune escape of bladder cancer, B7-H1 expression in 50 cases of bladder cancer was detected by using immunohistochemical method. Survival curves were con- structed using the Kaplan-Meier method and independent prognostic factors were evaluated using the Cox regression model. Our results showed that the positive rate of B7-H1 immunostaining in normal bladder tissue and bladder cancer was 0 and 72% respectively. The expression of B7-H1 was strongly associated with the pathological grade, clinical stage and recurrence （P〈0.05）. The survival rate was significantly lower in patients with B7-H1 positive group than in those with B7-H1 negative group and multi-variable analysis revealed that B7-H1 could be regarded as an independent factor in evaluating the prognosis of bladder cancer. It is concluded that the expression of B7-H1 is strongly associated with neoplastic progression and prognosis of bladder cancer. The manipulation of B7-H1 may become a beneficial target for immunotherapy in human bladder cancer.

B7 homologue 3 as a prognostic biomarker and potential therapeutic target in gastrointestinal tumors

The most common digestive system(DS)cancers,including tumors of the gastrointestinal tract(GIT)such as colorectal cancer(CRC),gastric cancer(GC)and esophageal cancer(EC)as well as tumors of DS accessory organs such as pancreatic and liver cancer,are responsible for more than one-third of all cancerrelated deaths worldwide,despite the progress that has been achieved in anticancer therapy.Due to these limitations in treatment strategies,oncological research has taken outstanding steps towards a better understanding of cancer cell biological complexity and heterogeneity.These studies led to new molecular target-driven therapeutic approaches.Different in vivo and in vitro studies have revealed significant expression of B7 homologue 3(B7-H3)among the most common cancers of the GIT,including CRC,GC,and EC,whereas B7-H3 expression in normal healthy tissue of these organs was shown to be absent or minimal.This molecule is able to influence the biological behavior of GIT tumors through the various immunological and nonimmunological molecular mechanisms,and some of them are shown to be the result of B7-H3-related induction of signal transduction pathways,such as Janus kinase 2/signal transducer and activator of transcription 3,phosphatidylinositol 3-kinase/protein kinase B,extracellular signal-regulated kinase,and nuclear factor-κB.B7-H3 exerts an important role in progression,metastasis and resistance to anticancer therapy in these tumors.In addition,the results of many studies suggest that B7-H3 stimulates immune evasion in GIT tumors by suppressing antitumor immune response.Accordingly,it was observed that experimental depletion or inhibition of B7-H3 in gastrointestinal cancers improved antitumor immune response,impaired tumor progression,invasion,angiogenesis,and metastasis and decreased resistance to anticancer therapy.Finally,the high expression of B7-H3 in most common cancers of the GIT was shown to be associated with poor prognosis.In this review,we summarize the established data from different GIT cancer-related studies and suggest that the B7-H3 molecule could be a promising prognostic biomarker and therapeutic target for anticancer immunotherapy in these tumors.

Comprehensive analysis of the protein phosphatase 2A regulatory subunit B56ε in pan-cancer and its role and mechanism in hepatocellular carcinoma

BACKGROUND B56εis a regulatory subunit of the serine/threonine protein phosphatase 2A,which is abnormally expressed in tumors and regulates various tumor cell functions.At present,the application of B56εin pan-cancer lacks a comprehensive analysis,and its role and mechanism in hepatocellular carcinoma(HCC)are still unclear.The Cancer Genome Atlas,Genotype-Tissue Expression,Gene Expression Profiling Interactive Analysis,and Tumor Immune Estimation Resource databases were used to analyze B56εexpression,prognostic mutations,somatic copy number alterations,and tumor immune characteristics in 33 tumors.The relationships between B56εexpression levels and drug sensitivity,immuno-therapy,immune checkpoints,and human leukocyte antigen(HLA)-related genes were further analyzed.Gene Set Enrichment Analysis(GSEA)was performed to reveal the role of B56εin HCC.The Cell Counting Kit-8,plate cloning,wound healing,and transwell assays were conducted to assess the effects of B56εinterference on the malignant behavior of HCC cells.RESULTS In most tumors,B56εexpression was upregulated,and high B56εexpression was a risk factor for adrenocortical cancer,HCC,pancreatic adenocarcinoma,and pheochromocytoma and paraganglioma(all P<0.05).B56εexpression levels were correlated with a variety of immune cells,such as T helper 17 cells,B cells,and macro-phages.There was a positive correlation between B56εexpression levels with immune checkpoint genes and HLA-related genes(all P<0.05).The expression of B56εwas negatively correlated with the sensitivity of most chemotherapy drugs,but a small number showed a positive correlation(all P<0.05).GSEA analysis showed that B56εexpression was related to the cancer pathway,p53 downstream pathway,and interleukin-mediated signaling in HCC.Knockdown of B56εexpression in HCC cells inhibited the proliferation,migration,and invasion capacity of tumor cells.Core Tip:The expression of protein phosphatase 2A(PP2A)subunit B56εis up-regulated in most tumors,and its high expression is a risk factor for adrenocortical cancer,hepatocellular carcinoma(HCC),pancreatic adenocarcinoma,and pheochromocytoma and paraganglioma.B56εexpression levels correlate with immune cells,immune checkpoint genes,human leukocyte antigen-related genes,and the sensitivity of chemotherapy drugs.In HCC,B56εexpression is related to the cancer pathway.Knockdown of B56εexpression in HCC cells can inhibit the proliferation,migration and invasion capacity of tumor cells.Our study supports PP2A subunit B56εas a prognostic marker and potential therapeutic target for HCC.

Experimental Study on the Antitumor Effect of Mouse B7-1 Gene

Summary: Mouse B7 1 cDNA was cloned by RT PCR from BALB/C mouse splenic cells and inserted into pcDNA3 to construct an eukaryotic expression vector. This constructor was named pCD mB7 1, in which the B7 1 cDNA was identified to be consistent with the data from other researchers. pCD mB7 1 plasmid was transfected into B16(F0) cells, and effective expression of mB7 1 in these tumor cells could be detected till the 6th month by RT PCR and RNA hybridization. Specific cytotoxity assay of lymphocytes was conducted after culturing with tumor cells and the results demonstrated that B16 cells transfected with B7 1 gene were more effective than B16 wt and B16 neo in inducing specific cytotoxity of lymphocytes against B16 wt cells. It is suggested that expression of B7 1 gene in tumor cells could enhance the immunogenicity and induce the effective antitumor immunity.

B细胞在荷瘤小鼠PD-(L)1抗体治疗中的作用

目的探讨肿瘤浸润性B细胞对程序性死亡受体(配体)1[programmed death(ligand)1,PD-(L)1]抑制剂治疗效果的影响,阐明B细胞影响免疫治疗的潜在机制。方法通过使用公共数据库中的黑色素瘤患者免疫治疗队列,分析B细胞与无进展生存时间(progression-free survival,PFS)以及免疫治疗响应的关系;使用TC-1和B16-OVA肿瘤细胞对6~8周龄雌性C57BL/6小鼠的皮下及肝脏进行荷瘤,构建肿瘤模型。比较B细胞清除对PD-(L)1抗体治疗效果的影响;对第15天的TC-1肿瘤微环境(tumor microenvironment,TME)进行流式细胞术检测,明确T细胞的数目、功能及表型变化;通过流式细胞术及实时荧光定量聚合酶链式反应(quantitative real-time polymerase chain reaction,qPCR)检测B细胞表面分子以及细胞因子。结果基于ICBatlas公共数据库中的ERP105482数据分析,肿瘤中CD19高表达的黑色素瘤患者PFS较低表达者更长(753 d vs 95 d,HR=0.3,95%CI:0.13~0.65,P=0.003)。B细胞在免疫治疗应答的患者中显著富集(P=0.01)。在小鼠TC-1肝脏荷瘤模型中,PD-(L)1抗体治疗后肿瘤质量明显减小(P<0.01),而B细胞的清除削弱了治疗效果。B细胞通过促进T细胞浸润和功能来增强PD-(L)1抗体治疗,且治疗后B细胞亚群发生变化,表现为PD-1低表达亚群增加(P<0.01)。结论PD-(L)1抗体治疗后,B细胞亚群中PD-1表达的下降可能是B细胞促进PD-(L)1抗体治疗效果的潜在机制。

HBV-HCC中HBx与免疫微环境的交互作用

乙型肝炎病毒X蛋白(HBx)是乙型肝炎病毒X基因(HBX)将自身DNA整合至人基因组,进而合成的多功能蛋白。HBX基因的表达受肝细胞免疫、微环境和机体免疫的监视和调控,其表达的蛋白也可通过激活肝星状细胞、参与机体免疫调节、调控炎性细胞因子和诱导细胞外基质重塑等参与肝细胞癌(HCC)抑制性免疫微环境的形成。HBx与免疫微环境的相互作用是影响乙肝病毒相关肝细胞癌(HBV-HCC)发生、发展的主要因素之一。深入研究HBx与免疫微环境相互作用机制,探索促进HBV-HCC抑制性免疫微环境形成的机制,有助于开发新型抗HCC药物,改善患者预后。本文就HBx与HBV-HCC免疫微环境的研究进展进行综述。

小鼠肺癌B_7疫苗细胞FLB_(2C)诱导的抗肿瘤免疫应答

目的 研究小鼠肺癌细胞与活化B淋巴细胞融合的疫苗细胞FLB2C 诱导细胞免疫反应情况。方法 用母本细胞LA795作对照 ,采用体外混合淋巴细胞培养 ,观察FLB2C 细胞刺激T细胞增殖情况 ;通过CTLL细胞MTT法测定FLB2C刺激T细胞产生分泌IL 2的作用 ;用FLB2C免疫小鼠 ,观察疫苗体内诱导小鼠CTL活性的能力。结果 FLB2C细胞在体外刺激T细胞增殖的作用明显比LA795强 ;FLB2C能刺激T细胞分泌IL 2 ,而LA795则不能 ;FLB2C细胞在体内能诱导CTL活性 ,其诱导的CTL在体外对FLB2C 细胞和LA795的杀伤率分别为 3 4%和 2 5 .3 % ,而LA795诱导的CTL对FLB2C和LA795杀伤率分别为 10 .5 %和 12 .2 5 % ,两者的杀伤率有显著性差异。结论 FLB2C细胞在体内外均能刺激T细胞免疫应答 ,其能力明显强于未融合的LA795小鼠肺癌细胞。将其作为疫苗细胞将有可能通过提高机体免疫应答而达到治疗肺癌的效果。

慢性乙型肝炎患者外周血单核细胞TLR7 mRNA表达与血清IL-12,HBV-DNA载量的相关性

目的:研究TLR7在慢性乙型肝炎患者外周血单核细胞的表达及其与IL-12,HBV-DNA载量之间的关系,探讨TLR7在抗病毒免疫应答中的作用机制及意义.方法:用实时荧光定量PCR法检测38例具有不同HBV-DNA载量的慢性乙型肝炎患者和12例正常对照者外周血单核细胞TLR7mR-NA表达,患者按病毒载量分为低、中、高3组.实时荧光定量PCR法测定血清病毒载量,ELISA法测定血清IL-12的水平,多组均数间的比较用方差分析,直线相关性分析用Spearman秩相关.结果:患者组中TLR7mRNA水平比正常对照组显著降低(P<0.01),3组患者间TLR7mRNA表达有显著差异且与血清HBV-DNA载量呈负相关,患者组血清IL-12比正常人显著增高,3组患者外周血清IL-12,低、中病毒载量组的差异无统计学意义(P>0.05),其余各组间差异有统计学意义(P<0.01),且与病毒载量呈负相关(r=-0.075,P<0.01).结论:TLR7在慢性乙型肝炎患者外周血单核细胞的表达下降,TLR7与IL-12可能在慢性乙型肝炎中对病毒清除有重要作用,其水平降低可能与疾病的慢性化机制有关.

Wilson disease associated with immune thrombocytopenia: A case report and review of the literature

BACKGROUND Wilson disease (WD) is a genetic disorder of hepatic copper excretion,leading to copper accumulation in various tissues.The manifestations are quite variable,and hemolytic anemia is the most common hematological presentation.WD associated with thrombocytopenia is very rare.CASE SUMMARY We report the case of an 11-year-old Chinese girl with WD that was associated with immune thrombocytopenia (ITP).Thrombocytopenia was the initial chief complaint for her to visit a hematologist,and ITP was diagnosed based on the results of a bone marrow biopsy and positive antiplatelet autoantibodies.About two weeks before the thrombocytopenia was found,the patient developed drooling.Tremors developed in her right hand about one week after being diagnosed with ITP,after which she was admitted to our hospital.Further evaluations were performed.Ceruloplasmin was decreased,with an increased level of copper in her 24-h urine excretion.Kayser Fleischer's ring (K-F ring) was positive.The ultrasound showed liver cirrhosis,and brain magnetic resonance imaging showed that the lenticular nucleus,caudate nucleus,and brainstem presented a low signal intensity in T1-weighted images and high signal intensity in T2-weighted images.WD was diagnosed and a genetic analysis was performed.A compound heterozygous mutation in ATP7B was detected;c.2333G>T (p.Arg778Leu) in exon 8 and c.3809A>G (p.Asn1270Ser) in exon 18.The former was inherited from her father and the latter from her mother.However,her parents showed normal liver function and negative K-F rings.Such a compound mutation in a case of WD associated with ITP in children has not been published previously.CONCLUSION WD can associate with thrombocytopenia but the mechanism is still unclear.We recommend that antiplatelet autoantibodies should be tested in WD patients with thrombocytopenia in future to verify the association.

Immunological classification of hepatitis B virus-positive hepatocellular carcinoma by transcriptome analysis

BACKGROUND Hepatitis B virus(HBV)infection is a major factor responsible for HBV+hepatocellular carcinoma(HCC).AIM An immunological classification of HBV+HCC may provide both biological insights and clinical implications for this disease.METHODS Based on the enrichment of 23 immune signatures,we identified two immunespecific subtypes(Imm-H and Imm-L)of HBV+HCC by unsupervised clustering.We showed that this subtyping method was reproducible and predictable by analyzing three different datasets.RESULTS Compared to Imm-L,Imm-H displayed stronger immunity,more stromal components,lower tumor purity,lower stemness and intratumor heterogeneity,lower-level copy number alterations,higher global methylation level,and better overall and disease-free survival prognosis.Besides immune-related pathways,stromal pathways(ECM receptor interaction,focal adhesion,and regulation of actin cytoskeleton)and neuro-related pathways(neuroactive ligand-receptor interaction,and prion diseases)were more highly enriched in Imm-H than in Imm-L.We identified nine proteins differentially expressed between Imm-H and Imm-L,of which MYH11,PDCD4,Dvl3,and Syk were upregulated in Imm-H,while PCNA,Acetyl-a-Tubulin-Lys40,ER-α_pS118,Cyclin E2,andβ-Catenin were upregulated in Imm-L.CONCLUSION Our data suggest that“hot”tumors have a better prognosis than“cold”tumors in HBV+HCC and that“hot”tumors respond better to immunotherapy.

调节性B细胞与肿瘤免疫逃逸

近年来,调节性B细胞(Breg)在肿瘤免疫研究中备受瞩目,多种实体肿瘤中均发现其大量浸润,直接或间接影响抗肿瘤免疫功能,发挥免疫调节效应。然而肿瘤浸润Breg的表型特征及其复杂的功能和机制尚有待系统解析。在此,基于现有研究成果,本文对肿瘤浸润Breg的表型、作用模式、驱动因素及临床相关性进行论述。

关于B 7-H 5/CD 28 H信号途径在结直肠癌中的表达以及对抗肿瘤免疫的影响分析

目的 通过检测B 7-H 5在不同分期的结直肠癌组织中的表达差异,研究B 7-H 5与肿瘤分期的关系,并通过比较肿瘤浸润细胞对不同表达程度的肿瘤细胞的杀伤试验,探索B 7-H 5在抗肿瘤免疫中的作用.方法 纳入健康志愿者、T1-T4期结直肠癌患者各20例,比较其外周血及组织中B 7-H 5的表达情况;提取组织中的肿瘤浸润细胞,在体外进行扩增,检测其对不同B 7-H 5表达水平肿瘤细胞的杀伤活性.结果 健康人外周血中无B 7-H 5,肿瘤分级越高,血液及组织中B 7-H 5的表达越高,提示B 7-H 5可以作为肿瘤严重程度的潜在指标.肿瘤浸润细胞杀伤试验表明,B 7-H 5是肿瘤杀伤的抑制因素.结论 血液及组织中B 7–H 5的表达含量可作为肿瘤分期及肿瘤免疫治疗疗效预测的潜在指标,但因本试验样本量较小,仍需要进一步的验证.

VCAN,expressed highly in hepatitis B virus-induced hepatocellular carcinoma,is a potential biomarker for immune checkpoint inhibitors

BACKGROUND As a proteoglycan,VCAN exists in the tumor microenvironment and regulates tumor proliferation,invasion,and metastasis,but its role in hepatocellular carcinoma(HCC)has not yet been elucidated.AIM To investigate the expression and potential mechanism of action of VCAN in HCC.METHODS Based on The Cancer Genome Atlas Liver Hepatocellular Carcinoma dataset,we explored the correlation between VCAN expression and clinical features,and analyzed the prognosis of patients with high and low VCAN expression.The potential mechanism of action of VCAN was explored by Gene Ontology analysis,Kyoto Encyclopedia of Genes and Genomes analysis,and gene set enrichment analysis.We also explored immune cell infiltration,immune checkpoint gene expression,and sensitivity of immune checkpoint[programmed cell death protein 1(PD-1)/cytotoxic T lymphocyte antigen 4(CTLA4)]inhibitor therapy in patients with different VCAN expression.VCAN mRNA expression and VCAN methylation in peripheral blood were tested in 100 hepatitis B virus(HBV)-related patients(50 HCC and 50 liver cirrhosis).RESULTS VCAN was highly expressed in HCC tissues,which was associated with a poor prognosis in HCC patients.No significant difference was found in VCAN mRNA expression in blood between patients with HBV-related cirrhosis and those with HCC,but there was a significant difference in VCAN methylation between the two groups.The correlation between VCAN and infiltrations of several different tumor immune cell types(including B cells,CD8+T cells,and eosinophils)was significantly different.VCAN was strongly related to immune checkpoint gene expression and tumor mutation burden,and could be a biomarker of sensitivity to immune checkpoint(PD1/CTLA4)inhibitors.In addition,VCAN mRNA expression was associated with hepatitis B e antigen,HBV DNA,white blood cells,platelets,cholesterol,and coagulation function.CONCLUSION High VCAN level could be a possible biomarker for poor prognosis of HCC,and its immunomodulatory mechanism in HCC warrants investigation.

包虫抗原B通过肿瘤坏死因子受体2调控巨噬细胞极化对小鼠免疫性血小板减少症的改善作用

目的:探讨包虫抗原B (HA-B)对小鼠免疫性血小板减少症(ITP)的改善作用,阐明其相关作用机制。方法:将野生型(WT)和肿瘤坏死因子受体2 (TNFR2)基因敲除(TNFR2^(-/-))的C57/B6小鼠分为WT对照组、WT-ITP模型组、WT-HA-B组、TNFR2^(-/-)对照组、TNFR2^(-/-)ITP模型组和TNFR2^(-/-)HA-B组,检测各组小鼠体质量、脏器指数和血常规指标,采用流式细胞术检测各组小鼠外周血中M2巨噬细胞百分率,酶联免疫吸附试验(ELISA)法检测各组小鼠血清中精氨酸酶1(Arg1)、白细胞介素10 (IL-10)、诱导型一氧化氮合成酶(iNOS)和白细胞介素6 (IL-6)水平,采用实时荧光定量PCR (RT-qPCR)法检测各组小鼠骨髓来源巨噬细胞(BMDM)中Arg1、IL-10、iNOS和IL-6表达水平。分别将WT对照组和TNFR2^(-/-)对照组小鼠BMDM诱导为M2巨噬细胞(WT M2组和TNFR2^(-/-)M2组),加入HA-B (WT M2+HA-B组和TNFR2^(-/-)M2+HA-B组),采用RT-qPCR法检测各组细胞中Arg1和IL-10mRNA表达水平。结果:分别与WT对照组和TNFR2^(-/-)对照组比较,WT-ITP模型组和TNFR2^(-/-)ITP模型组小鼠体质量降低(P<0.05),脾脏和胸腺指数升高(P<0.05),血小板和红细胞数量减少(P<0.05),血红蛋白水平降低(P<0.05),白细胞数量增加(P<0.05),凝血时间延长(P<0.05);外周血中M2巨噬细胞百分率降低(P<0.05),血清中Arg1和IL-10水平降低(P<0.05),iNOS和IL-6水平升高(P<0.05);BMDM中Arg1和IL-10 mRNA表达水平降低(P<0.05),iNOS和IL-6 mRNA表达水平升高(P<0.05)。与WT-ITP模型组比较,WT-HA-B组小鼠体质量增加(P<0.05),脾脏和胸腺指数降低(P<0.05),血小板和红细胞数量增加(P<0.05),血红蛋白水平升高(P<0.05),白细胞数量减少(P<0.05),凝血时间缩短(P<0.05);外周血中M2巨噬细胞百分率升高(P<0.05),血清中Arg1和IL-10水平升高(P<0.05),iNOS和IL-6水平降低(P<0.05);BMDM中Arg1和IL-10 mRNA表达水平升高(P<0.05),iNOS和IL-6 mRNA表达水平降低(P<0.05)。与WT-HA-B组比较,TNFR2^(-/-)HA-B组小鼠体质量降低(P<0.05),脾脏和胸腺指数升高(P<0.05),血小板和红细胞数量减少(P<0.05),血红蛋白水平降低(P<0.05),白细胞数量增加(P<0.05),凝血时间延长(P<0.05);外周血中M2巨噬细胞百分率降低(P<0.05),血清中Arg1和IL-10水平降低(P<0.05),iNOS和IL-6水平升高(P<0.05);BMDM中Arg1和IL-10mRNA表达水平降低(P<0.05),iNOS和IL-6 mRNA表达水平升高(P<0.05)。与WT M2组比较,WT M2+HA-B组M2巨噬细胞中Arg1和IL-10 mRNA表达水平升高(P<0.05);与WT M2+HA-B组比较,TNFR2^(-/-)M2+HA-B组M2巨噬细胞中Arg1和IL-10 mRNA表达水平降低(P<0.05)。结论:HA-B可通过TNFR2促进巨噬细胞M2极化,进而发挥治疗ITP的作用。