B7-H6在恶性肿瘤中的研究进展

B7-H6是新近发现的免疫球蛋白超家族成员,是NK细胞表面活化性受体NKP30的膜结合配体.B7-H6较特异地表达于多种肿瘤组织,并且其表达水平与多种临床病理参数及预后显著相关.细胞表面B7-H6胞外域脱落后形成可溶性B7-H6,下调NKP30介导的NK细胞毒性及细胞因子的分泌.研究发现B7-H6通过蛋白酶抑制剂、My...     

B7-H1 and B7-H3 are independent predictors of poor prognosis in patients with non-small cell lung cancer

B7-H1 and B7-H3, two members of the B7 family that are thought to regulate T-cell activation, are expressed in human non-small cell lung cancer (NSCLC). However, their prognostic significance is poorly understood. In the present study we reported that B7-H1 and B7-H3 were expressed in 96/128 (72.7%) and 89/128 (69.5%) samples, respectively. B7-H1 and B7-H3 expression and the number of infiltrating T-cell intracellular antigen-1+ and interferon-γ+ cells in NSCLC tissues were significantly higher than those in the adjacent tissues (p<0.01). High B7-H1 or B7-H3 expression was associated with lymph node metastasis and TNM stage (p<0.05, respectively). Sex, TNM stage, B7-H1, B7-H3, and T-cell intracellular antigen-1 expression remained significant prognostic factors after adjusting for other prognostic factors in a multivariate Cox proportional hazards regression model. In vitro studies revealed that knockdown of B7-H3 on tumor cells enhanced T-cell growth and interferon-γ secretion when stimulated by anti-CD3 and anti-CD28 monoclonal antibodies. Interferon-γ reduced CXCR4 expression on cancer cells and inhibited the CXCL12-induced cell migration. B7-H1 and B7-H3 are independent predictors of poorer survival in patients with NSCLC. Interference of the signal pathways of these negative regulatory molecules might be a new strategy for treating NSCLC.     

协同刺激分子PD-L1、B7-H3与B7-H4在卵巢癌中的表达及其意义

协同刺激分子PD-L1、B7-H3与B7-H4可与T细胞及其受体结合并抑制T细胞的增殖和过度活化,在细胞免疫应答过程中起重要的调控作用,已被多项研究证明与肿瘤的免疫原性及肿瘤的发生、发展密切相关。这3种分子在正常卵巢组织中均不表达,而在卵巢癌组织中呈不同程度的高表达,它们可能在促进卵巢癌的发生、转变及病情进展过程中起重要作用,研究其作用机制对卵巢癌的早期诊断及靶向治疗有一定的意义。     

Programmed Death-Ligand 1 Heterogeneity and Its Impact on Benefit From Immune Checkpoint Inhibitors in NSCLC

IntroductionProgrammed death-ligand 1 (PD-L1) expression may vary in different disease sites and at different time points of the disease course. We aimed to investigate PD-L1 heterogeneity and its usefulness as a predictive value for immune checkpoint inhibitor (ICI) therapy in patients with NSCLC.MethodsPD-L1 expression was analyzed in 1398 patients with NSCLC. The predictive value of PD-L1 for ICIs in 398 patients with metastatic NSCLC was assessed.ResultsPD-L1 was significantly associated with biopsy sites (p = 0.004). Adrenal, liver, and lymph node (LN) metastases had the highest PD-L1 expression as a continuous variable and at 1% or 50% cutoff. PD-L1 expression was lower in bone and brain metastases. Among 112 patients with two specimens tested, 55 (49%) had major changes in PD-L1 falling into different clinically relevant categories (<1%, 1%–49%, ≥50%) at different time points. Previous ICI therapy was associated with significant decrease in PD-L1 compared with treatment-naive counterparts (p = 0.015). Patients with metastatic NSCLC treated with ICI (n = 398) were divided into three cohorts on the basis of biopsy sites: lung (n = 252), LN (n = 85), and distant metastasis (n = 61). Higher PD-L1 in lung or distant metastasis specimens was associated with higher response rate, longer progression-free survival, and overall survival. However, PD-L1 in LN biopsies was not associated with either response or survival.ConclusionsPD-L1 varies substantially across different anatomical sites and changes during the clinical course. PD-L1 from different biopsy sites may have different predictive values for benefit from ICIs in NSCLC.     

负性协同刺激分子B7-H4在非小细胞肺癌组织中的表达及其临床意义

目的：探讨负性协同刺激分子B7H4在原发性非小细胞肺癌（nonsmall cell lung cancer,NSCLC）组织中的表达及其与NSCLC临床病理特征间的关系。方法：选取南通大学第三附属医院胸外科2008年1月至2009年4月手术切除并经病理证实的NSCLC标本52例。免疫组织化学法检测NSCLC组织中B7H4分子的表达及CD3+T细胞浸润程度,分析B7H4表达水平与NSCLC组织中CD3+T细胞浸润、临床病理特征间的关系。结果：52例NSCLC组织中B7H4表达阳性率为4808%（25/52）,正常肺组织不表达或较少表达B7H4,差异具有统计学意义（P<0.05）。B7H4表达水平与NSCLC临床分期和淋巴结转移呈正相关（P<0.05）,与CD3+T淋巴细胞浸润程度呈负相关（P<0.05）,与其他临床病理参数无关（P>005）。结论：负性协同刺激分子B7H4在NSCLC发病过程中可能起重要的作用,其阳性表达与NSCLC临床分期及淋巴结转移密切相关,B7H4为NSCLC的诊断及治疗提供参考依据。     

Clinical Benefit From Immunotherapy in Patients With SCLC Is Associated With Tumor Capacity for Antigen Presentation

IntroductionA small percentage of patients with SCLC experience durable responses to immune checkpoint blockade (ICB). Defining determinants of immune response may nominate strategies to broaden the efficacy of immunotherapy in patients with SCLC. Prior studies have been limited by small numbers or concomitant chemotherapy administration.MethodsCheckMate 032, a multicenter, open-label, phase 1/2 trial evaluating nivolumab alone or with ipilimumab was the largest study of ICB alone in patients with SCLC. We performed comprehensive RNA sequencing of 286 pretreatment SCLC tumor samples, assessing outcome on the basis of defined SCLC subtypes (SCLC-A, -N, -P, and -Y), and expression signatures associated with durable benefit, defined as progression-free survival more than or equal to 6 months. Potential biomarkers were further explored by immunohistochemistry.ResultsNone of the subtypes were associated with survival. Antigen presentation machinery signature (p = 0.000032) and presence of more than or equal to 1% infiltrating CD8+ T cells by immunohistochemistry (hazard ratio = 0.51, 95% confidence interval: 0.27–0.95) both correlated with survival in patients treated with nivolumab. Pathway enrichment analysis revealed the association between durable benefit from immunotherapy and antigen processing and presentation. Analysis of epigenetic determinants of antigen presentation identified LSD1 gene expression as a correlate of worse survival outcomes for patients treated with either nivolumab or the combination of nivolumab and ipilimumab.ConclusionsTumor antigen processing and presentation is a key correlate of ICB efficacy in patients with SCLC. As antigen presentation machinery is frequently epigenetically suppressed in SCLC, this study defines a targetable mechanism by which we might improve clinical benefit of ICB for patients with SCLC.     

Programmed Death-Ligand 1 Copy Number Alteration as an Adjunct Biomarker of Response to Immunotherapy in Advanced NSCLC

IntroductionThis study aimed to evaluate the value of programmed death-ligand 1 (PD-L1) copy number (CN) alteration as an additional biomarker to standard immunohistochemistry (IHC) in predicting response to immune checkpoint inhibitor (ICI) therapy in advanced NSCLC.MethodsBefore ICI monotherapy, tumor PD-L1 CN alteration (gain, neutral, or loss) was called using whole-exome sequencing data and compared with IHC results (tumor proportion score ≥50, 1–49, or 0). Progression-free survival (PFS) and overall survival were correlated with both biomarkers. In addition, the impact of CN alteration was further evaluated in two independent cohorts using next-generation sequencing panel.ResultsA total of 291 patients with advanced-stage NSCLC met the study inclusion criteria. Although the IHC classification distinguished the best responsive group (tumor proportion score ≥ 50), the CN-based classification distinguished the worst responsive group (CN loss) from the others (PFS, p = 0.020; overall survival, p = 0.004). After adjusting for IHC results, CN loss was an independent risk factor for progression (adjusted hazard ratio = 1.32, 95% confidence interval: 1.00–1.73, p = 0.049) and death (adjusted hazard ratio = 1.39, 95% confidence interval: 1.05–1.85, p = 0.022). A risk classification system was developed on the basis of IHC and CN profiles, which outperformed the conventional IHC system. In the validation cohorts, CN loss determined by next-generation sequencing panel was independently associated with worse PFS after ICI treatment, revealing its practical value.ConclusionsThis is the first study to directly compare CN alterations with IHC results and survival outcomes after anti–PD-(L)1 therapy. Tumor PD-L1 CN loss can serve as an adjunct biomarker to predict the lack of response. Prospective studies are required to further validate this biomarker.     

负性共刺激分子B7-H1和B7-H4在结直肠癌中的表达及其临床意义

目的:检测结直肠癌组织中负性共刺激分子B7-H1和B7-H4的表达、T细胞亚群的浸润情况,探讨其临床意义。方法:收集苏州大学附属第四医院2003年1月至2003年12月50例结直肠癌患者的癌组织标本以及5例患者的癌旁组织标本,免疫组织化学法检测结直肠癌组织中B7-H1和B7-H4的表达以及T细胞亚群的浸润,分析B7-H1、B7-H4的表达与结直肠癌患者临床病理特征及T细胞浸润的相关性,分析B7-H1、B7-H4的表达和CD3+T、CD8+T淋巴细胞浸润程度与患者预后的相关性。结果:结直肠癌组织高表达B7-H1(44%)和B7-H4(56%),而癌旁组织不表达(P<0.01)。B7-H1在结肠癌组织中的表达较直肠癌显著升高(P<0.05);随着Duke’s分期的升高,B7-H4的表达水平也呈上升趋势(P<0.05)。结直肠癌组织中B7-H1的表达与CD3+T细胞浸润呈负相关(P<0.05),但与B7-H4的表达无关。B7-H1的表达水平与患者预后呈负相关(P<0.05),且B7-H1和B7-H4同时高表达的患者总体生存率显著降低(P<0.05)。结论:负性共刺激分子B7-H1和B7-H4在人结直肠癌组织中高表达,并与患者总体生存率相关,两者的共同检测对结直肠癌诊断和预后判断具有一定的临床价值。     

Clinical significance and regulation of the costimulatory molecule B7-H1 in pancreatic cancer

We investigated the expression pattern and clinical significance of the costimulatory ligands B7-1, B7-2, B7-H1, and B7-DC, and their counter-receptors CTLA-4 and PD-1 in pancreatic cancer. Gene expression of all examined costimulatory molecules was significantly upregulated in pancreatic cancer tissues. B7-1, B7-2, B7-H1, and B7-DC protein was detectable in pancreatic cancer cells. Only the expression of B7-H1 significantly correlated with postoperative survival (p<0.0001). B7-H1 was inducible in cultured pancreatic cancer cells by IFN-gamma and significantly correlated with the level of IFN-gamma expression in human pancreatic cancer tissues (Spearman rho=0.4536,p=0.0029). B7-H1 positive tumors showed an increased prevalence of tumor-infiltrating regulatory T cells (T(regs)) compared to B7-H1 negative tumors. Among the investigated costimulatory molecules only tumor-associated B7-H1 seems to be of prognostic relevance in pancreatic cancer. B7-H1 might, therefore, be involved in the downregulation of antitumor responses through regulation of T(regs) in pancreatic cancer. Our findings also suggest a dual role of IFN-gamma in antitumor response. Through induction of B7-H1 in pancreatic cancer cells IFN-gamma might contribute to the evasion of antitumor immunity.     

YES1 Is a Druggable Oncogenic Target in SCLC

IntroductionSCLC is an extremely aggressive subtype of lung cancer without approved targeted therapies. Here we identified YES1 as a novel targetable oncogene driving SCLC maintenance and metastasis.MethodsAssociation between YES1 levels and prognosis was evaluated in SCLC clinical samples. In vitro functional experiments for proliferation, apoptosis, cell cycle, and cytotoxicity were performed. Genetic and pharmacologic inhibition of YES1 was evaluated in vivo in cell- and patient-derived xenografts and metastasis. YES1 levels were evaluated in mouse and patient plasma-derived exosomes.ResultsOverexpression or gain/amplification of YES1 was identified in 31% and 26% of cases, respectively, across molecular subgroups, and was found as an independent predictor of poor prognosis. Genetic depletion of YES1 dramatically reduced cell proliferation, three-dimensional organoid formation, tumor growth, and distant metastasis, leading to extensive apoptosis and tumor regressions. Mechanistically, YES1-inhibited cells revealed alterations in the replisome and DNA repair processes, that conferred sensitivity to irradiation. Pharmacologic blockade with the novel YES1 inhibitor CH6953755 or dasatinib induced marked antitumor activity in organoid models and cell- and patient-derived xenografts. YES1 protein was detected in plasma exosomes from patients and mouse models, with levels matching those of tumors, suggesting that circulating YES1 could represent a biomarker for patient selection/monitoring.ConclusionsOur results provide evidence that YES1 is a new druggable oncogenic target and biomarker to advance the clinical management of a subpopulation of patients with SCLC.     

Programmed Death Ligand 1 Immunohistochemistry in Triple-Negative Breast Cancer: Evaluation of Inter-Pathologist Concordance and Inter-Assay Variability

PurposeThe programmed death ligand 1 (PD-L1) SP142 assay with a 1% immune cell (IC) cutoff is approved for the selection of advanced triple-negative breast cancer (TNBC) patients for atezolizumab treatment. We aimed to evaluate the interobserver concordance of PD-L1 scoring and inter-assay variability of various PD-L1 assays in TNBC.MethodsThirty patients with primary TNBC were selected, and SP142, SP263, 22C3, and E1L3N assays were performed. PD-L1 staining in ICs and tumor cells (TCs) was scored by 10 pathologists who were blinded to the assay. The interobserver concordance among pathologists and the inter-assay variability of the four PD-L1 assays were analyzed. For SP142, the intraobserver concordance among the six pathologists was analyzed after training.ResultsThe adjusted means of PD-L1 IC scoring ranged from 6.2% to 12.9% for the four assays; the intraclass correlations showed moderate (0.584–0.649) reader concordance. The PD-L1 IC scoring with a 1% cutoff resulted in identical scoring in 40.0%–66.7% of cases and a poor to moderate agreement (Fleiss κ statistic [FKS] = 0.345–0.534) for the four assays. The SP142 assay had the widest range of positive rate (56.5%–100.0%), lowest number of cases with identical scoring, and lowest FKS at 1% cutoff. Pairwise comparison of adjusted means showed significantly decreased PD-L1 staining in SP142 compared with the other assays in both ICs and TCs. As for the intraobserver concordance in the SP142 assay, the overall percent agreement was 87.8% with a 1% IC cutoff. After training, the proportion of cases with identical scoring at a 1% IC cutoff increased to 70.0%; the FKS also increased to 0.610.ConclusionThe concordance of PD-L1 IC scoring among pathologists was low, at the 1% cutoff for the SP142 assay without training. SP142 showed the lowest PD-L1 expression in both IC and TC.     

Beyond Steroids: Immunosuppressants in Steroid-Refractory or Resistant Immune-Related Adverse Events

IntroductionThe optimal management for immune-related adverse events (irAEs) in patients who do not respond or become intolerant to steroids is unclear. Guidelines suggest additional immunosuppressants on the basis of case reports and expert opinion.MethodsWe evaluated patients with lung cancers at Memorial Sloan Kettering Cancer Center treated with immune checkpoint blockade from 2011 to 2020. Pharmacy records were queried to identify patients who received systemic steroids and an additional immunosuppressant (e.g., tumor necrosis factor-α inhibitor, mycophenolate mofetil). Patient records were manually reviewed to evaluate baseline characteristics, management, and outcomes.ResultsAmong 2750 patients with lung cancers treated with immune checkpoint blockade, 51 (2%) received both steroids and an additional immunosuppressant for a severe irAE (tumor necrosis factor-α inhibitor (73%), mycophenolate mofetil (20%)). The most common events were colitis (53%), pneumonitis (20%), hepatitis (12%), and neuromuscular (10%). At 90 days after the start of an additional immunosuppressant, 57% were improved from their irAE, 18% were unchanged, and 25% were deceased. Improvement was more common in hepatitis (five of six) and colitis (18 of 27) but less common in neuromuscular (one of five) and pneumonitis (3 of 10). Of the patients who died, 8 of 13 were attributable directly to the irAE and 4 of 13 were related to toxicity from immunosuppression (three infection-related deaths, one drug-induced liver injury leading to acute liver failure).ConclusionsSteroid-refractory or resistant irAEs events are rare. Although existing treatments help patients with hepatitis and colitis, many patients with other irAEs remain refractory or experience toxicities from immunosuppression. A more precise understanding of the pathophysiology of specific irAEs is needed to guide biologically-informed treatments for severe irAEs.     

Classification of Tumor Immune Microenvironment According to Programmed Death-Ligand 1 Expression and Immune Infiltration Predicts Response to Immunotherapy Plus Chemotherapy in Advanced Patients With NSCLC

IntroductionAccording to mechanisms of adaptive immune resistance, tumor immune microenvironment (TIME) is classified into four types: (1) programmed death-ligand 1 (PD-L1)–negative and tumor-infiltrating lymphocyte (TIL)–negative (type I); (2) PD-L1–positive and TIL-positive (type II); (3) PD-L1–negative and TIL-positive (type III); and (4) PD-L1–positive and TIL-negative (type IV). However, the relationship between the TIME classification model and immunotherapy efficacy has not been validated by any large-scale randomized controlled clinical trial among patients with advanced NSCLC.MethodsOn the basis of RNA-sequencing and immunohistochemistry data from the ORIENT-11 study, we optimized the TIME classification model and evaluated its predictive value for the efficacy of immunotherapy plus chemotherapy.ResultsPD-L1 mRNA expression and immune score calculated by the ESTIMATE method were the strongest predictors for the efficacy of immunotherapy plus chemotherapy. Therefore, they were determined as the optimized definition of the TIME classification system. When compared between combination therapy and chemotherapy alone, only the type II subpopulation with high immune score and high PD-L1 mRNA expression was significantly associated with improved progression-free survival (PFS) (hazard ratio = 0.12, 95% confidence interval: 0.06–0.25, p < 0.001) and overall survival (hazard ratio = 0.27, 95% confidence interval: 0.13–0.55, p < 0.001). In the combination group, the type II subpopulation had a much longer survival time, not even reaching the median PFS or overall survival, but the other three subpopulations were susceptible to having similar PFS. In the chemotherapy group, there was no marked association between survival outcomes and TIME subtypes.ConclusionsOnly patients with both high PD-L1 expression and high immune infiltration could benefit from chemotherapy plus immunotherapy in first-line treatment of advanced NSCLC. For patients lacking either PD-L1 expression or immune infiltration, chemotherapy alone might be a better treatment option to avoid unnecessary toxicities and financial burdens.     

B7-H1蛋白在人胰腺癌组织中的表达及临床意义

背景与目的：恶性肿瘤细胞表达共刺激分子B7-H1来抑制T细胞功能,逃避机体的抗肿瘤免疫反应。本研究通过检测人胰腺癌组织中B7-H1蛋白的表达,探讨其与胰腺癌患者临床病理指标及预后之间的关系。方法：采用免疫组化法检测B7-H1在40例胰腺癌组织和10例癌旁正常胰腺组织中的表达情况,用卡方检验或确切概率法分析B7-H1的表达与胰腺癌患者各临床病理指标以及生存时间的关系。结果：40例胰腺癌组织中B7-H1阳性率为45.00%（18/40）,高于癌旁正常胰腺组织（P〈0.05）;B7-H1表达与患者肿瘤分期、术前CA19-9有关（P〈0.05）。多因素Cox回归模型显示,B7-H1表达是胰腺癌患者无复发生存期、总生存期的独立危险因素。结论：人胰腺癌组织中存在B7-H1蛋白表达,B7-H1表达与胰腺癌患者的预后有关。     

B7-H3的作用机制及其在非小细胞肺癌中的研究新进展

肺癌是目前世界上癌症相关性死亡的主要原因之一。其中最常见的类型为非小细胞肺癌（non-small cell lung cancer,NSCLC）。肺癌患者预后不佳,因此探索新型治疗方案对提高患者预后来说是非常必要的。据研究,共刺激分子B7-H3（CD276）在抗肿瘤免疫中起着重要的作用,而免疫系统的调节机制在抑制肿瘤微环境中的抗肿瘤T细胞介导的免疫反应中起着重要作用。本文阐述了B7-H3的作用机制及其在非小细胞肺癌微环境中表达的临床意义。     

协同刺激分子B7-H4在食管癌前病变组织中的表达及临床意义

目的：研究协同刺激分子B7同系物4（B7-H4）在人食管癌前病变组织中的表达变化,探讨其在食管鳞状细胞癌形成中的临床意义。方法：收集内镜切除的食管组织标本58例,其中正常组织16例,低级别上皮内瘤变（LGIN）23例,高级别上皮内瘤变（HGIN）19例。通过苏木素-伊红（HE）染色评价食管组织病理变化。分别采用实时荧光定量PCR（qPCR）检测B7-H4 mRNA在食管组织的表达;免疫组织化学和Western blot检测B7-H4蛋白在食管病变组织中的表达;酶联免疫吸附法（ELISA）检测食管组织细胞因子IL-6、IL-10和IFN-γ的表达,并分析各细胞因子与B7-H4蛋白表达的相关性。结果：免疫组化和HE染色结果提示,B7-H4表达与食管组织病理分级呈显著正相关（P < 0.01）。qPCR结果提示,与正常食管组织相比,食管癌前病变组织B7-H4 mRNA表达无显著改变（P > 0.05）。而Western blot实验结果显示,与正常食管组织相比,食管癌前病变组织B7-H4蛋白表达显著升高（P < 0.05）。Westernblot和ELISA实验结果显示B7-H4表达与IL-6表达呈正相关（P < 0.01）。结论：随着食管癌前病变病理分级的提高,B7-H4蛋白的表达逐渐上调,B7-H4可能与IL-6相互作用促进了食管鳞状细胞癌的发生发展。