Expression and clinical value of programmed cell death-ligand 1(PD-L1)in diffuse large B cell lymphoma:a retrospective study

Background: The programmed cell death-1(PD-1)/programmed cell death-ligand 1(PD-L1) pathway inhibits the activation of T cells and plays a crucial role in the negative regulation of cellular and humoral immune responses.Diffuse large B-cell lymphoma(DLBCL) is the most common lymphoid malignancy in adults. In the present study, we aimed to detect the expression of PD-L1 in DLBCL and to analyze its relationship with prognosis.Methods: We reviewed medical records of 204 newly diagnosed DLBCL patients in Sun Yat-sen University Cancer Center between October 2005 and August 2012. The expression of PD-L1 in tumor tissues from these 204 patients was detected using immunohistochemical(IHC) assay. The expression of anaplastic lymphoma kinase(ALK), CD5,CD30, and C-Myc in tumor specimens from 109 patients was detected using IHC, and Epstein-Barr virus(EBV)-encoded RNAs(EBERs) were detected using fluorescence in situ hybridization. The Spearman method was used for correlation analysis. The Kaplan-Meier method with log-rank test was used for univariate analysis. Cox proportional hazards model was used for multivariate analysis.Results: Of the 204 patients, 100(49.0%) were PD-L1-positive in tumor cells and 44(21.6%) were PD-L1-positive in tumor microenvironment. PD-L1 expression in tumor cells and tumor microenvironment were more common in the non-germinal center B-cell-like(GCB) subtype than in the GCB subtype(P = 0.02 and P= 0.04). Patients with PD-L1 expression in tumor microenvironment were more likely to be resistant to first-line chemotherapy when compared with the patients without PD-L1 expression in tumor microenvironment(P = 0.03). PD-L1 expression in tumor microenvironment was negatively correlated with C-Myc expression(r =-0.20, P = 0.04). No correlations were detected between PD-L1 expression and the expression of ALK, CD5, and CD30 as well as EBERs. The 5-year overall survival(OS)rates were 50.0% and 67.3% in patients with and without PD-L1 expression in tumor cells(P = 0.02). PD-L1 expression in tumor cells was an independent risk predictor for OS(P < 0.01).Conclusions: PD-L1 expression is more common in the non-GCB subtype than in the GCB subtype. PD-L1 expression in tumor microenvironment has a negative correlation with C-Myc. PD-L1 positivity predicts short survival in DLBCL patients. For patients with PD-L1 expression, more strategy such as anti-PD-L1 antibody treatment should be recommended.

Locoregional radiotherapy in patients with distant metastases of nasopharyngeal carcinoma at diagnosis

Systemic chemotherapy is the basic palliative treatment for metastatic nasopharyngeal carcinoma(NPC); however, it is not known whether locoregional radiotherapy targeting the primary tumor and regional lymph nodes affects the survival of patients with metastatic NPC. Therefore, we aimed to retrospectively evaluate the benefits of locoregional radiotherapy. A total of 408 patients with metastatic NPC were included in this study. The mortality risks of the patients undergoing supportive treatment and those undergoing chemotherapy were compared with that of patients undergoing locoregional radiotherapy delivered alone or in combination with chemotherapy. Univariate and multivariate analyses were conducted. The contributions of independent factors were assessed after adjustment for covariates with significant prognostic associations (P<0.05). Both locoregional radiotherapy and systemic chemotherapy were identified as significant independent prognostic factors of overall survival(OS). The mortality risk was similar in the group undergoing locoregional radiotherapy alone and the group undergoing systemic chemotherapy alone [multi-adjusted hazard ratio(HR)=0.9, P=0.529]; this risk was 60% lower than that of the group undergoing supportive treatment(HR=0.4, P=0.004) and 130% higher than that of the group undergoing both systemic chemotherapy and locoregional radiotherapy(HR=2.3, P<0.001). In conclusion, locoregional radiotherapy, particularly when combined with systemic chemotherapy, is associated with improved survival of patients with metastatic NPC.

Prognostic significance of the pre-chemotherapy lymphocyte-to-monocyte ratio in patients with previously untreated metastatic colorectal cancer receiving FOLFOX chemotherapy

Background:As a surrogate marker of systemic inflammation,the lymphocyte-to-monocyte ratio(LMR) is an independent prognostic factor for various malignancies.This study investigated the prognostic significance of the pre-chemotherapy LMR in patients with previously untreated metastatic colorectal cancer(mCRC) receiving chemotherapy.Methods:The present study included newly diagnosed mCRC patients treated between January 2005 and December 2013 with FOLFOX chemotherapy,specifically oxaliplatin 180 mg/m^2 on day 1,with leucovorin 400 mg/m^2administered as a 2-hour infusion before the administration of 5-fluorouracil 400 mg/m^2 as an intravenous bolus injection,and 5-fluorouracil 2400 mg/m^2 as a 46-h infusion immediately after 5-fluorouracil bolus injection.The LMR was calculated as the absolute count of lymphocytes divided by the absolute count of monocytes.COX proportional hazards analysis was performed to evaluate the association of LMR with survival outcomes.Results:A total of 488 patients were included.Patients with high pre-chemotherapy LMR experienced significant improvements in progression-free survival(PFS,9.2 vs.7.6 months,P < 0.001) and overall survival(OS,19.4 vs.16.6 months,P < 0.001) compared with patients with low pre-chemotherapy LMR.Subsequent COX multivariate analysis showed that high pre-chemotherapy LMR(>3.11) was an independent favorable prognostic factor for PFS and OS.Additionally,patients whose LMR remained high(high-high subgroup),increased(low-high subgroup),or decreased(high-low subgroup) following chemotherapy showed better results in terms of PFS and OS than patients whose LMR remained low(low-low subgroup) after chemotherapy.Conclusions:For patients with previously untreated mCRC receiving FOLFOX chemotherapy,an elevated pre-chemotherapy LMR is an independent favorable prognostic factor for PFS and OS,and changes in the LMR before and after chemotherapy seem to predict the benefit of chemotherapy.

New risk factors and new tendency for central nervous system relapse in patients with diffuse large B-cell lymphoma:a retrospective study

Background: In patients with difuse large B?cell lymphoma(DLBCL), central nervous system(CNS) relapse is uncom?mon but is nearly always fatal. This study aimed to determine the risk factors for CNS relapse in DLBCL patients and to evaluate the eicacy of rituximab and intrathecal chemotherapy prophylaxis for CNS relapse reduction.Methods: A total of 511 patients with newly diagnosed DLBCL treated at the Sun Yat?sen University Cancer Center between January 2003 and December 2012 were included in the study. Among these patients, 376 received R?CHOP regimen(rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone) as primary treatment, and 135 received CHOP regimen(cyclophosphamide, doxorubicin, vincristine, and prednisone) as primary treatment. Intrathe?cal chemotherapy prophylaxis(methotrexate plus cytarabine) was administered to those who were deemed at high risk for CNS relapse. In the entire cohort and in the R?CHOP set in particular, the Kaplan–Meier method coupled with the log?rank test was used for univariate analysis, and the Cox proportional hazards model was used for multivariate analysis. Diferences were evaluated using a two?tailed test, and P < 0.05 was considered signiicant.Results: At a median follow?up of 46 months, 25(4.9%) patients experienced CNS relapse. There was a trend of reduced occurrence of CNS relapse in patients treated with rituximab; the 3?year cumulative CNS relapse rates were 7.1% in CHOP group and 2.7% in R?CHOP group(P = 0.045). Intrathecal chemotherapy prophylaxis did not confer much beneit in terms of preventing CNS relapse. Bone involvement [hazard ratio(HR) = 4.21, 95% conidence interval(CI) 1.38–12.77], renal involvement(HR = 3.85, 95% CI 1.05–14.19), alkaline phosphatase(ALP) >110 U/L(HR = 3.59, 95% CI 1.25–10.34), serum albumin(ALB) <35 g/L(HR = 3.63, 95% CI 1.25–10.51), treatment with rituxi?mab(HR = 0.34, 95% CI 0.12–0.96), and a time to complete remission ≤ 108 days(HR = 0.22, 95% CI 0.06–0.78) were independent predictive factors for CNS relapse in the entire cohort. Bone involvement(HR = 4.44, 95% CI 1.08–18.35), bone marrow involvement(HR = 11.70, 95% CI 2.24–60.99), and renal involvement(HR = 10.83, 95% CI 2.27–51.65) were independent risk factors for CNS relapse in the R?CHOP set.Conclusions: In the present study, rituximab decreased the CNS relapse rate of DLBCL, whereas intrathecal chemo?therapy prophylaxis alone was not suicient for preventing CNS relapse. Serum levels of ALB and ALP, and the time to complete remission were new independent predictive factors for CNS relapse in the patients with DLBCL. In the patients received R?CHOP regimen, a trend of increased CNS relapse was found to be associated with extranodal lesions.

Use of subsequent PET/CT in diffuse large B-cell lymphoma patients in complete remission following primary therapy

Interim 18F-fluorodeoxyglucose(FDG) positron emission tomography/computed tomography(I-PET/CT) is a powerful tool for monitoring the response to therapy in diffuse large B-cell lymphoma(DLBCL). This retrospective study aimed to determine when and how to use I-PET/CT in DLBCL. A total of 197 patients treated with rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone(R-CHOP) were enrolled between October 2005 and July 2011; PET/CT was performed at the time of diagnosis(PET/CT0), after 2 and 4 cycles of chemotherapy(PET/CT2 and PET/CT4, respectively), and at the end of treatment(F-PET/CT). According to the International Harmonization Project for Response Criteria in Lymphoma, 110 patients had negative PET/CT2 scans, and 87 had positive PET/CT2 scans. The PET/CT2-negative patients had significantly higher 3-year progression-free survival rate(75.8% vs. 38.2%) and 3-year overall survival rate(93.5% vs. 55.6%) than PET/CT2-positive patients. All PET/CT2-negative patients remained negative at PET/CT4, but 3 were positive at F-PET/CT. Among the 87 PET/CT2-positive patients, 57 remained positive at F-PET/CT, and 32 progressed during chemotherapy(15 at PET/CT4 and 17 at F-PET/CT). Comparing PET/CT4 with PET/CT0, 7 patients exhibited progression, and 8 achieved partial remission. Comparing F-PET/CT with PET/CT0, 10 patients exhibited progression, and 7 achieved partial remission. In conclusion, our results indicate that I-PET/CT should be performed after 2 rather than 4 cycles of immunochemotherapy in DLBCL patients. There is a limited role for subsequent PET/CT in the detection of relapse in PET/CT2-negative patients, but repeat PET/CT is required if the PET/CT2 findings are positive.

Qualitative visual trichotomous assessment improves the value of fluorine-18 fluorodeoxyglucose positron emission tomography/computed tomography in predicting the prognosis of diffuse large B-cell lymphoma

Introduction:Fluorine-18 fluorodeoxyglucose(18F-FDG)positron emission tomography/computed tomography(PET/CT)is a powerful tool for monitoring the response of diffuse large B-cell lymphoma(DLBCL)to therapy,but the criteria to interpret PET/CT results remain under debate.We investigated the value of post-treatment PET/CT in predicting the prognosis of DLBCL patients when interpreted according to qualitative visual trichotomous assessment(QVTA)criteria compared with the Deauvil e criteria.Methods:In this retrospective study,final PET/CT scans of DLBCL patients treated with rituximab-based regimens between October 2005 and November 2010 were interpreted using the Deauvil e and QVTA criteria.Survival curves were estimated using Kaplan-Meier analysis and compared using the log-rank test.Results:A total of 253 patients were enrol ed.The interpretation according to the Deauvil e criteria revealed that 181patients had negative PET/CT scan results and 72 had positive results.The 3 year overal survival(OS)rate was significantly higher in patients with negative scan results than in those with positive results(91.6%vs.57.5%,P<0.001).The72 patients with positive scan results according to the Deauville criteria were divided into two groups by the interpretation according to the QVTA criteria:29 had indeterminate results,and 43 had positive results.The 3 year OS rate was significantly higher in patients with indeterminate scan results than in those with positive results(91.2%vs.33.5%,P<0.001)but was similar between patients with negative and indeterminate scan results(91.6%vs.91.2%,P=0.921).Conclusions:Compared with the Deauvil e criteria,using the QVTA criteria for interpreting post-treatment PET/CT scans of DLBCL patients is likely to reduce the number of false positive results.The QVTA criteria are feasible for therapeutic outcome evaluation and can be used to guide risk-adapted therapy.

潜伏膜蛋白1介导IL-2Rα上调促进NK/T细胞淋巴瘤的形成和化疗耐药

背景与目的 NK/T细胞淋巴瘤(natural killer/T-cell lymphoma,NKTCL)是一种高度侵袭性的非霍奇金淋巴瘤,经常发生化疗耐药。可溶性白细胞介素-2受体α(IL-2 receptorα,IL-2Rα)在NKTCL患者中血清水平升高,并与疗效和生存期显著相关。本研究旨在通过代表性细胞系中过表达IL-2Rα来探讨IL-2Rα的潜在作用。方法在人NK细胞系NK-92和NKTCL细胞系SNK-6中检测IL-2Rα的水平。使用慢病毒载体在NK-92细胞中表达潜伏膜蛋白1(latent membrane protein 1,LMP1),以及在NK-92和SNK-6细胞中表达IL-2Rα,分析这些基因在增殖、凋亡、细胞周期分布和化疗敏感性中的生物学功能。结果 IL-2Rα在SNK-6细胞中的表达水平显著高于NK-92细胞。在NK-92细胞中表达LMP1可以显著上调IL-2Rα水平,而MAPK/NF-κB途径相关蛋白的选择性抑制剂可以显著下调IL-2Rα。在SNK-6细胞中IL-2Rα的过表达通过改变细胞周期分布促进细胞增殖,并诱导产生对吉西他滨、多柔比星和门冬酰胺酶的耐药,这些效应可以被抗IL-2Rα抗体逆转。结论我们的研究显示,在NKTCL细胞中LMP1激活MAPK/NF-κB途径,从而上调IL-2Rα表达。IL-2Rα过表达促进NKTCL的生长和化疗耐药,表明该白细胞介素受体可作为潜在的治疗靶点。

细胞程序性死亡配体-1(PD-L1)在弥漫大B细胞淋巴瘤中的表达及临床价值的回顾性研究

背景与目的细胞程序性死亡受体-1(programmed cell death-1,PD-1)/细胞程序性死亡配体-1(programmed cell death-ligand 1,PD-L1)通路会抑制T细胞的活化,在细胞和体液免疫应答的负调控中起关键作用。弥漫大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)是成年人中最为常见的淋巴样恶性肿瘤,本研究旨在检测DLBCL中PD-L1的表达,并分析其与预后的关系。方法回顾分析2005年10月至2012年8月204例在中山大学肿瘤防治中心新诊断为DLBCL的患者病历资料,采用免疫组织化学(immunohistochemical,IHC)法检测204例患者肿瘤组织中PD-L1的表达。应用IHC检测109例患者肿瘤组织中的间变性淋巴瘤激酶(anaplastic lymphoma kinase,ALK)、CD5、CD30和C-Myc的表达,应用荧光原位杂交技术检测EB病毒(Epstein-Barr virus,EBV)编码RNAs(EBV-encoded RNAs,EBERs)的表达。应用Spearman方法进行相关分析,采用具有对数秩检验的Kaplan-Meier法进行单因素分析,采用Cox比例风险模型进行多因素分析。结果在选取的204例患者中,100例(49.0%)患者肿瘤细胞中PD-L1呈阳性,44例(21.6%)患者肿瘤微环境中PD-L1呈阳性。与生发中心B细胞样(germinal center B-cell-like,GCB)亚型相比,肿瘤细胞和肿瘤微环境中表达PD-L1在非生发中心B细胞样亚型中更为常见(P=0.02和P=0.04)。与肿瘤微环境中不表达PD-L1的患者相比,肿瘤微环境中PD-L1表达的患者更容易对一线化疗产生耐药性(P=0.03)。肿瘤微环境中PD-L1表达与C-Myc表达呈负相关(r=-0.20,P=0.04),而PD-L1表达与ALK、CD5、CD30和EBERs的表达无相关性。此外,肿瘤细胞中有PD-L1表达和无PD-L1表达的患者的5年总生存期(overall survival,OS)分别为50.0%和67.3%(P=0.02)。肿瘤细胞中的PD-L1表达是OS的独立风险预测因素(P<0.01)。结论PD-L1在非GCB亚型中的表达比在GCB亚型中更常见。肿瘤微环境中PD-L1的表达与C-Myc的表达呈负相关。PD-L1阳性预示DLBCL患者生存期短。因此对于PD-L1表达的患者,应推荐更多的治疗策略,如抗PD-L1抗体治疗。

BRD2 induces drug resistance through activation of the RasGRP1/Ras/ERK signaling pathway in adult T-cell lymphoblastic lymphoma

Background:Adult patients with T-cell lymphoblastic lymphoma(T-LBL)are treated with high-intensity chemotherapy regimens,but the response rate is still unsatisfactory because of frequent drug resistance.We aimed to investigate the potential mechanisms of drug resistance in adults with T-LBL.Methods:Gene expression microarray was used to identify differential mRNA expression profiles between chemotherapy-resistant and chemotherapy-sensitive adult T-LBL tissues.Real-time PCR and immunohistochemistry were performed to detect the expression of bromodomain-containing protein 2(BRD2)and c-Myc in fresh-frozen T-LBL tissues from 85 adult patients.The Ras pull-down assay was performed to monitor Ras activation.Chromatin immunoprecipitation assays were used to analyze the binding of E2F transcription factor 1(E2F1)/BRD2 to the RAS guanyl releasing protein 1(RasGRP1)promoter region.The drug resistance effect and mechanism of BRD2 were determined by both in vivo and in vitro studies.Results:A total of 86 chemotherapy resistance-related genes in adult T-LBL were identified by gene expression microarray.Among them,BRD2 was upregulated in chemotherapy-resistant adult T-LBL tissues and associated with worse progressionfree survival and overall survival of 85 adult T-LBL patients.Furthermore,BRD2 suppressed doxorubicin(Dox)-induced cell apoptosis both in vitro and in vivo.The activation of RasGRP1/Ras/ERK signaling might contribute to the Dox resistance effect of BRD2.Besides,OTX015,a bromodomain and extra-terminal(BET)inhibitor,reversed the Dox resistance effect of BRD2.Patient-derived tumor xenograft demonstrated that the sequential use of OTX015 after Dox showed superior therapeutic effects.Conclusions:Our data showed that BRD2 promotes drug resistance in adult T-LBL through the RasGRP1/Ras/ERK signaling pathway.Targeting BRD2 may be a novel strategy to improve the therapeutic efficacy and prolong survival of adults with TLBL.

IL-2Rα up-regulation is mediated by latent membrane protein 1 and promotes lymphomagenesis and chemotherapy resistance in natural killer/T-cell lymphoma

Background:Natural killer/T-cell lymphoma(NKTCL)is a highly aggressive non-Hodgkin lymphoma often resistant to chemotherapy.Serum level of soluble IL-2 receptorα(IL-2Rα)is elevated in NKTCL patients and correlates signifi-cantly with treatment response and survival.In the current study we examined the potential role of IL-2Rαby over-expressing IL-2Rαin representative cell lines.Methods:Levels of IL-2Rαwere evaluated in the human natural killer cell line NK-92 and the NKTCL cell line SNK-6.Lentiviral vectors were used to express latent membrane protein 1(LMP1)in NK-92 cells,and IL-2Rαin both NK-92 and SNK-6 cells.The biological effects of these genes on proliferation,apoptosis,cell cycle distribution,and chemosensitiv-ity were analyzed.Results:Expression of IL-2Rαwas significantly higher in SNK-6 cells than in NK-92 cells.Expressing LMP1 in NK-92 cells remarkably up-regulated IL-2Rαlevels,whereas selective inhibitorss of the proteins in the MAPK/NF-κB pathway significantly down-regulated IL-2Rα.IL-2Rαoverexpression in SNK-6 cells promoted cell proliferation by altering cell cycle distribution,and induced resistance to gemcitabine,doxorubicin,and asparaginase.These effects were reversed by an anti-IL-2Rαantibody.Conclusions:Our results suggest that LMP1 activates the MAPK/NF-κB pathway in NKTCL cells,up-regulating IL-2Rαexpression.IL-2Rαoverexpression promotes growth and chemoresistance in NKTCL,making this interleukin receptor a potential therapeutic target.