Hypermethylation of PRDM1/Blimp‐1 promoter in extranodal NK/T‐cell lymphoma,nasal type: an evidence of predominant role in its downregulation

The loss of PRDM1 expression is common in extranodal NK/T‐cell lymphoma, nasal type (EN‐NK/T‐NT), but the role of promoter methylation in silencing PRDM1 expression remains unclear. Hence, we performed pyrosequencing analysis to evaluate the promoter methylation of PRDM1 gene in vivo and in vitro, to analyze the association between methylation and its expression, and to assess cellular effects of PRDM1 reexpression. The promoter hypermethylation of PRDM1 gene was detected in 11 of 25 EN‐NK/T‐NT cases (44.0%) and NK92 and NKL cells. The promoter hypermethylation of PRDM1 was significantly correlated with PRDM1 expression in vivo and in vitro, predominantly contributing to the loss of PRDM1 expression compared with genetic deletion and aberrant expression of miR‐223 in EN‐NK/T‐NT. PRDM1 expression was significantly restored by demethylation treatment, which induced cell proliferation suppression, cell cycle arrest, and apoptosis increase. We also found that PRDM1 reexpression could downregulate the expression of Ets‐1, T‐bet, granzyme B, and c‐myc. Our findings demonstrated that the promoter hypermethylation of PRDM1 harbored a predominant role in the downregulation of PRDM1 expression, significantly affecting the biological behavior of tumor cells in EN‐NK/T‐NT.     

结外NK/T细胞淋巴瘤发生的分子机制研究进展

结外NK/T细胞淋巴瘤(extranodal NK/T cell lymphoma,ENKTCL) 是我国继弥漫大B淋巴细胞淋巴瘤之后第二常见的非霍奇金淋巴瘤(non-hadgkin lymphoma,NHL).结外NK/T细胞淋巴瘤恶性程度高、临床进展快、预后差,对其发病机制的研究尤为重要.本文综述了近年来结外NK/T细胞淋巴瘤发生的分子机制研究进展,主要涉及该淋巴瘤的常见基因突变、NF-κB信号通路及JAK-STAT信号通路的主要激活机制.     

结外鼻型NK/T细胞淋巴瘤的研究进展

结外鼻型NK／T细胞淋巴瘤是鼻腔淋巴瘤中较为常见、恶性程度高且预后较差的病种。因此．了解其发生发展、临床表现、病理诊断、治疗及预后是非常重要的。     

Clinical study of l‐asparaginase in the treatment of extranodal NK/T‐cell lymphoma,nasal type

Extranodal natural killer/T‐cell lymphoma, nasal type, (ENKTL) is a rare distinct entity of non‐Hodgkin lymphoma. It is prevalent in Asia and Latin America but rare in North America and Europe. ENKTL represents an aggressive clinical course and a poor prognosis especially for advanced disease. There is no standard chemotherapeutic regimen for ENKTL. Recently, the efficacy of l ‐asparaginase in ENKTL has been confirmed. A series of l ‐asparaginase‐containing chemotherapeutic regimens have been studied in clinical trials and have significantly improved the efficacy and prognosis for patients with ENKTL. This review will focus on pharmacology of l ‐asparaginase, the efficacy of a series of l ‐asparaginase‐containing regimens in the treatment of ENKTL and future clinical study directions of l ‐asparaginase‐containing regimens in ENKTL. Copyright © 2015 John Wiley & Sons, Ltd.     

Treatment outcome of front-line systemic chemotherapy for localized extranodal NK/T cell lymphoma in nasal and upper aerodigestive tract

We analysed the treatment outcome of localized extranodal NK/T cell lymphoma initially treated with CEOP-B chemotherapy based on the primary site of involvement (nasal cavity vs. upper aerodigestive tract) and treatment modality (chemotherapy vs. chemotherapy followed by radiotherapy. Forty-three patients newly diagnosed as extranodal NK/T cell lymphoma were analysed: 29 cases from nasal cavity/nasopharynx and 14 from upper aerodigestive tract. Twenty-six patients were treated with chemotherapy alone, while adjuvant radiotherapy was given to 17 patients. Overall response rate to front-line CEOP-B chemotherapy was 67.4% (29/43) and the complete remission (CR) rate was 44.2% (19/43). Median overall and disease-free survival was 26.87 months [95% confidence interval (CI) = 8.71 - 45.03] and 15.27 months (95% CI = 2.92 - 27.62). The responders (CR or partial response) to initial CEOP-B chemotherapy showed longer overall survival than non-responders (P = 0.0026). Local relapse was observed to be higher in the chemotherapy alone group compared to the chemoradiotherapy group. Adjuvant radiotherapy failed to improve survival; thus, the median disease-free survival of the chemotherapy and chemoradiotherapy groups was not different (P = 0.9101). There may be a tendency for better overall survival in group of upper aerodigestive tract lymphoma than the nasal cavity/nasopharynx group (P = 0.0643). However, front-line CEOP-B chemotherapy has a limited role and adjuvant radiotherapy failed to improve survival in localized extranodal NK/T cell lymphoma.     

Macrophage inflammatory protein 1 alpha (MIP‐1α) may be associated with poor outcome in patients with extranodal NK/T‐cell lymphoma

The macrophage inflammatory protein 1α (MIP‐1α) is anticipated to have a role in extranodal natural killer (NK)/T‐cell lymphoma (ENKTL) because the expression of MIP‐1α is related to Epstein–Barr virus (EBV) latency in EBV‐related non‐Hodgkin lymphoma cells. Thus, we measured the serum level of MIP‐1α in 69 patients with ENKTL using frozen serum samples that were archived at diagnosis. As serum level of MIP‐1α was not detectable in 19 patients (range: 0–24.37 pg/mL), patients were dichotomized into positive (n = 50) and negative (n = 19) MIP‐1α groups according to the presence of detectable level of MIP‐1α in serum. MIP‐1α‐positive group showed a significantly poor overall survival (OS) in comparison with the MIP‐1α‐negative group (p = 0.004). In the subgroup analysis, the positivity of MIP‐1α was significantly associated with OS in patients with stage IIIE/IV and a detectable level of EBV DNA (p = 0.002 and 0.032, respectively). Multivariate analysis also showed that the positivity of MIP‐1α was independently associated with worse OS together with bone marrow involvement (p = 0.002). An in vitro study with patient‐derived ENKTL tumour cells showed the expression of CCR1 and CCR5 on the surface of tumour cells (28% and 14%, respectively) , and the addition of MIP‐1α to the culture media of tumour cells increased cell growth supporting the negative impact of MIP‐1α on the prognosis of ENKTL patients. In conclusion, serum levels of MIP‐1α could predict survival outcomes in patients with ENKTL. Therefore, MIP‐1α should be considered for prognostication and a potential therapeutic target. Copyright © 2016 John Wiley & Sons, Ltd.     

Clinical development of anti‐CD19 chimeric antigen receptor T‐cell therapy for B‐cell non‐Hodgkin lymphoma

B‐cell non‐Hodgkin lymphoma (B‐NHL) is the most frequent hematological malignancy. Although refined chemotherapy regimens and several new therapeutics including rituximab, a chimeric anti‐CD20 monoclonal antibody, have improved its prognosis in recent decades, there are still a substantial number of patients with chemorefractory B‐NHL. Anti‐CD19 chimeric antigen receptor (CAR) T‐cell therapy is expected to be an effective adoptive cell treatment and has the potential to overcome the chemorefractoriness of B‐cell leukemia and lymphoma. Recently, several clinical trials have shown remarkable efficacy of anti‐CD19 CAR T‐cell therapy, not only in B‐acute lymphoblastic leukemia but also in B‐NHL. Nonetheless, there are several challenges to overcome before introduction into clinical practice, such as: (i) further refinement of the manufacturing process, (ii) further improvement of efficacy, (iii) finding the optimal infusion cell dose, (iv) optimization of lymphocyte‐depleting chemotherapy, (v) identification of the best CAR structure, and (vi) optimization of toxicity management including cytokine release syndrome, neurologic toxicity, and on‐target off‐tumor toxicity. Several ways to solve these problems are currently under study. In this review, we describe the updated clinical data regarding anti‐CD19 CAR T‐cell therapy, with a focus on B‐NHL, and discuss the clinical implications and perspectives of CAR T‐cell therapy.     

Primary nasal lymphoma,NK/T‐cell type: Report of two cases with similar presentation but different outcome

Two cases of natural killer (NK)/T‐cell primary nasal lymphoma with similar clinical presentations are reported, for comparison and contrast, to highlight the clinical issues and challenges posed by this unusual disease, its aggressiveness being matched only by its rarity. Presenting as a lesion in the nasal cavity with histological features of malignant lymphoma, primary nasal lymphoma is an uncommon extranodal lymphoma, which poses problems in both diagnosis and management. In people of oriental descent, the common cell subtype is NK/T‐cell. Although it is generally thought that combination treatment with chemotherapy and radiation is the best management for early stage non‐Hodgkin's lymphoma (NHL), there is still debate as to whether combined therapy is optimal treatment for this particular subtype of NHL, given that it responds less well to conventional chemotherapy. Herein we report two patients to illustrate these controversies.     

培门冬酶治疗结外NK/T细胞淋巴瘤的临床疗效

目的：分析结外 NK/ T 细胞淋巴瘤行培门冬酶与左旋门冬酰胺酶治疗的临床疗效。方法：以2009年6月至2014年12月病理诊断为结外 NK/ T 细胞淋巴瘤的患者为研究对象。将患者随机分为2组,培门冬酶组和左旋门冬酰胺酶组,两组均采用放疗以及以铂类为基础的两联方案化疗。在治疗结束后评价并对比患者的近期、远期疗效以及毒副反应。结果：共搜集结外 NK/ T 细胞淋巴瘤患者171例,其中培门冬酶组86例,左旋门冬酰胺酶组85例,近期疗效比较,P <0.05,差异有统计学意义,3个月、6个月、1年生存率比较,经Logrank 检验,P =0.000<0.05,培门冬酶组的3个月、6个月、1年生存率显著高于左旋门冬酰胺酶组;两组患者均出现了不同程度的血液学毒性及恶心、呕吐等反应,恶心、呕吐、过敏、高血糖等发生率比较,P <0.05,差异有统计学意义;白细胞、血小板下降、肝功能损害的发生率比较,P >0.05。结论：培门冬酶较左旋门冬酰胺酶可明显提高结外 NK/ T 细胞淋巴瘤的近期疗效。不良反应少,安全性高。     

Epstein‐Barr virus‐negative extranodal “true” natural killer‐cell lymphoma harbouring a KDM6A mutation

Extranodal natural killer (NK)/T‐cell lymphoma, nasal type (ENKTL) is an extranodal aggressive T or NK‐cell lymphoma that is characteristically associated with Epstein‐Barr virus (EBV) infection and cytotoxic tissue‐destructive features. Although ENKTL is described as a distinct entity according to the 2008 WHO classification, a considerable complexity is associated with the differential diagnosis of other T‐cell lymphomas with respect to tumour cell origins, locations, and the presence of EBV infection, as well as molecular and cytogenetic abnormalities. Here, we report a rare case of EBV‐negative ENKTL, where the absence of EBV in the true NK‐lineage cells was confirmed by extensive phenotypic and genotypic analyses. Furthermore, using the next‐generation sequencing approach, we identified mutations in the tumour suppressor genes KDM6A and TP53. The clinicopathological characteristics were almost similar to those of EBV‐positive ENKTL, except for the absence of EBV and histologically apparent angioinvasiveness. This is the first reported ENKTL case with mutations in the KDM6A gene. KDM6A is one of the histone‐modifying genes that are mutated in many human diseases including haematological cancers. Epigenetic regulation of gene expression has recently been demonstrated in ENKTL, and a similar pathway is thought to play an oncogenic role in EBV‐negative ENKTL. Our report shows the extent of comprehensive examination required before making a definitive diagnosis for NK‐ and T‐cell neoplasms and broadens the therapeutic options for potential targets.     

结外鼻型NK/T细胞淋巴瘤的内科治疗进展

 结外鼻型自然杀伤（NK）/T细胞淋巴瘤是一种高度侵袭性非霍奇金淋巴瘤,因发病率低,缺乏大样本前瞻性随机对照研究,仍未确定标准一线化疗方案。因多药耐药基因1高表达,对蒽环类药物为主的常规化疗抗拒。近年来出现一些有效率较高的方案,含左旋门冬酰胺酶或吉西他滨的方案近期有效率高,可作为首选方案,而靶向治疗、免疫调节治疗和造血干细胞移植仍在探索之中。     

结外鼻型NK/T细胞淋巴瘤27例临床分析

 目的　分析结外鼻型NK／T细胞淋巴瘤（ENKL）的临床特征、不同治疗方法的疗效及影响预后的因素。方法　回顾性分析27例ENKL患者,予单纯放疗3例,单纯化疗9例,其余15例采用放化疗联合治疗,其中3例患者行自体造血干细胞移植（auto-HSCT）。对B组症状、乳酸脱氢酶（LDH）、一般状况评分、国际预后指数评分、Ann Arbor分期、治疗模式和近期疗效进行单因素分析。单因素分析采用 Kaplan-Meier法。结果　全组平均生存时间为32（2～42）个月。1、2、3年的总体生存（OS）率分别为79.5 %、71.6 %、53.7 %。早期（Ⅰ期+Ⅱ期）及晚期（Ⅲ期+Ⅳ期）患者2年生存率分别为83.3 %、62.3 %（P＝0.368）,早期患者单纯放疗或化疗4例均获总有效（OR）（CR+PR）,化疗联合放疗10例,OR 9例。晚期患者单纯化疗8例,OR 2例;化疗联合放疗5例,OR 2例。单因素分析显示,年龄及近期疗效是影响生存率的主要因素（P＜0.05）。结论　对于ENKL早期患者,放疗或化疗可取得较好的疗效,放化疗联合治疗及auto-HSCT是治疗晚期ENKL的重要方法。年龄及近期疗效可作为判断ENKL预后的参考因素。     

结外鼻型NK/T细胞淋巴瘤放化疗研究进展

结外鼻型NK/T细胞淋巴瘤（extranodal NK/T-cell lymphoma,nasal type ENKTL）是侵袭性非霍奇金淋巴瘤的一种特殊类型,由于发病率低,目前临床治疗证据大部分来自回顾性分析或小样本的Ⅱ期临床试验,缺乏大型的随机对照研究,暂无统一的治疗标准。放疗、化疗是ENKTL的主要治疗方法,但目前放、化疗策略选择仍存在争议性,是否联合放化疗、放化疗联合方式、化疗方案选择等方面均未形成统一的认识。对于早期（Ⅰ/Ⅱ期）患者目前多以放疗联合化疗的治疗方法,Ⅲ/Ⅳ期患者多采用以全身化疗为主。以L-门冬酰胺酶（L-ASP）为基础的化疗药物在各期及复发难治性ENKTL中疗效结果显示均较CHOP或CHOP样化疗方案好。最佳的化疗方案以及化疗与放疗结合方式仍需通过更多的、更大型的Ⅲ期随机对照试验来证实。寻找准确的预后因素进行风险分层,根据风险分层结果进行治疗是未来的研究热点和方向。本文将有关放、化疗的研究进展进行综述,以期对该病的治疗提供参考性指导。     

结外鼻型NK/T细胞淋巴瘤117例临床病理分析

 目的　分析结外鼻型NK／T细胞淋巴瘤（NKTL）的临床病理特点,探讨其与疾病预后的关系,为临床个体化治疗提供依据。方法　回顾117例确诊为NKTL患者的病理临床资料。病理诊断均采用常规组织学、免疫组织化学、T细胞重排和原位杂交方法检测EB病毒（EBV）编码小RNA（EBER）。EBV DNA拷贝数检测采用PCR方法。所有患者经过国际预后指数（IPI）和Ki-67等指标临床评估后均采用化疗和放疗联合治疗。随访患者并对临床指标与2年总生存（OS）率和无进展生存（PFS）率的关系进行单因素分析。结果　免疫组织化学结果提示各指标阳性率：CD3为90.6 %,CD56为94.0 %,CD45RO为92.9 %,TIA为97.9 %,Granzyme B为97.7 %,EBER为100.0 %。117例患者中位年龄为43.2岁（14～77岁）,原发鼻者95例（81.2 %）,Ki-67平均值为（48.3±2.6）%;原发非鼻者22例（18.8 %）,包括原发咽喉、舌、扁桃体、淋巴结、皮肤、肝、肠、中枢神经系统和睾丸。与原发鼻的NKTL患者相比,原发肝和肠者Ki-67值较高,且Ki-67值大于80 %的NKTL患者均在1年内死亡。Ki-67值为60 %～80 %的45例2年OS率为60.0 %,与Ki-67值为30 %～60 %的33例OS率（86.3 %）和Ki-67值小于30 %的8例OS率（100.0 %）分别相比,差异均有统计学意义（P＝0.047、0.011）。Ki-67值为60 %～80 %者2年PFS率为36.0 %,与Ki-67值为30 %～60 %者PFS率（57.5 %）相比,差异无统计学意义（P＝0.07）,与Ki-67值小于30 %者PFS率（78.0 %）相比,差异有统计学意义（P＝0.02）。CD56阴性的8例CD3表达阳性,TCR重排均为阳性,提示为T细胞来源,原发部位均为鼻部,无全身症状,2年OS率和PFS率分别为100.0 %和70.0 %,高于CD56阳性患者,差异均有统计学意义（P＝0.03、0.02）。13例检测了EBV DNA拷贝数,5例高于正常值者OS率为60.0 %,余8例OS率为100.0 %。结论　除了IPI外,NKTL原发部位和病理特征,尤其是Ki-67、CD56、EBER和EBV DNA拷贝数与其临床预后的关系不容忽视,也是NKTL预后判断的独立因素。     

鼻型结外NK/T细胞淋巴瘤的诊治策略

 【摘要】　鼻型结外NK／T细胞淋巴瘤是中国常见的非霍奇金淋巴瘤,其诊断和治疗仍存在不少难题。根据典型的组织学表现和CD+3ε、CD+56、EBER+可确定诊断,同时需检查与预后相关的指标。建议在正电子发射计算机断层显像-电子计算机X射线断层扫描技术（PET-CT）检查基础上仍采用Ann Arbor分期系统。早期患者宜化疗+放疗,晚期患者建议联合化疗,化疗可采用含有左旋门冬酰胺酶的方案。对造血干细胞移植应持积极态度,新治疗方案应在大样本随机对照研究中评价。     

Post-treatment plasma EBV-DNA positivity predicts early relapse and poor prognosis for patients with extranodal NK/T cell lymphoma in the era of asparaginase

Circulating Epstein-Barr virus (EBV) DNA is a biomarker of EBV-associated malignancies. Its prognostic value in early stage NK/T-cell lymphoma (NKTCL) in the era of asparaginase was investigated. 68 patients were treated with a median of 4 cycles of asparaginase-based chemotherapy followed by a median of 54.6Gy (range 50–60Gy) radiation. The amount of EBV-DNA was prospectively measured in both pretreatment and post-treatment plasma samples by real-time quantitative PCR. At the end of treatment, complete response (CR) rate was 79.4%, and overall response rate (ORR) was 88.2%. Patients with negative pretreatment EBV-DNA had a higher CR rate (96.0% vs. 69.8%, p = 0.023). The 3-year progression-free survival (PFS) rate and overall survival (OS) rate was 71% and 83%, respectively. In multivariate survival analysis, post-treatment EBV-DNA positivity and treatment response (non-CR) were prognostic factors for both worse PFS and OS (p < 0.05). Local tumor invasion was also a prognostic factor for worse OS (p = 0.010). In patients with CR, post-treatment EBV-DNA positivity correlated with inferior PFS and OS (both p < 0.0001). In patients with positive pretreatment EBV-DNA, negative post-treatment EBV-DNA correlated with better PFS and OS (both p < 0.0001). These findings indicate that post-treatment EBV-DNA positivity can predict early relapse and poor prognosis for patients with early stage NKTCL in the era of asparaginase, and may be used as an indicator of minimal residual disease.     

A phase 2 study of methotrexate,etoposide, dexamethasone,and pegaspargase chemotherapy for newly diagnosed,relapsed, or refractory extranodal natural killer/T‐cell lymphoma,nasal type: a multicenter trial in Northwest China

The nasal type of extranodal natural killer/T‐cell lymphoma is a rare aggressive lymphoma with poor prognosis. To discover a successful treatment, we investigated the efficacy and safety of chemotherapy with methotrexate, etoposide, dexamethasone, and polyethylene glycol‐asparaginase (MESA). Three cycles of MESA were administered to 46 patients with new or relapsed/refractory natural killer/T‐cell lymphoma. Complete response after 3 treatment cycles was 43.5%, the overall response rate was 87%, and 2‐year overall survival was 83.4%. Complete response was significantly better for newly diagnosed patients than for patients with relapsed/refractory disease. Patients with newly diagnosed disease had a significantly better overall response rate after 1, but not after 2 or 3 treatment cycles. Overall survival and progression‐free survival did not differ over 2 years. Grade 1/2 toxicities were frequent, but MESA was associated with fewer grade 3/4 events or treatment‐related deaths. These results will require confirmation in larger prospective trials.     

贫血在结外鼻型NK/T细胞淋巴瘤预后价值——中国淋巴瘤放疗协作组多中心研究

目的 分析贫血在结外鼻型NK/T细胞淋巴瘤中的预后价值。方法 回顾分析 1225例在中国10个医疗中心确诊并且接受首程治疗的结外鼻型NK/T细胞淋巴瘤。中国贫血标准:海平面地区成年男性Hb<120 g/L,成年女性<110 g/L。贫血程度分级:极重度Hb≤30 g/L,重度 31~60 g/L,中度 61~90 g/L,轻度>90 g/L。结果 全组患者就诊时有 199例伴有贫血,占16.2%。贫血患者具有更多预后不良因素,贫血患者中分期为Ⅱ—Ⅳ期、中位年龄>60岁、ECOG评分2—4分、NK/T细胞淋巴瘤预后指数≥2分的比例较高。贫血、非贫血患者 5年OS分别为49.4%、63.3%(P<0.01),5年PFS分别为35.4%、56.0%(P<0.01)。单因素分析显示贫血、年龄、ECOG评分、B组症状、乳酸脱氢酶、原发部位、原发肿瘤侵犯以及分期是OS和PFS影响因素,多因素分析显示贫血仍然是预后影响因素。结论 贫血在结外鼻型NK/T细胞淋巴瘤中少见,这部分患者预后较差,贫血是预后影响因素。     

自体外周血干细胞移植治疗鼻型结外NK/T细胞淋巴瘤

目的:探讨自体外周血干细胞移植(autologous peripheral blood stem cell transplantation,APBSCT)治疗鼻型结外NK/T细胞淋巴瘤的疗效。方法:在我院自2000年1月-2009年12月结束治疗的31例经病理形态学及免疫组织化学检查确诊的鼻型结外NK/T细胞淋巴瘤患者,经交替应用CHOP方案、VDLP方案和MEOP方案各2疗程化疗后,均应用直线加速器进行原发部位的三维适形放射治疗。随后经化疗联合重组人粒细胞集落刺激因子(rh G-CSF)方法动员自体外周血干细胞,经TBI联合VEMAC方案实施预处理后,进行自体外周血干细胞移植。随访观察时间为3-5年。结果:放化疗的主要不良反应为骨髓抑制及轻度局部黏膜损伤。全部患者均获得造血重建,无特殊并发症出现。随访1年时,治疗总有效率96.8%;随访3年时,无病存活率占全组患者的87.1%;随访5年时,无病存活患者占该组随访5年患者的81.5%。结论:自体外周血干细胞移植治疗鼻型结外NK/T细胞淋巴瘤可取得满意疗效。