vdac-1基因调控多发性骨髓瘤细胞表达髓系分化抗原CD33实验研究

目的:探讨线粒体外膜电压依赖阴离子通道-1(vdac-1)基因对多发性骨髓瘤(MM)细胞表达髓系分化抗原CD33的影响,为MM的治疗提供新的思路。方法:应用细胞转染技术将携带vdac-1基因的质粒转染不表达Pax5和CD45的骨髓瘤细胞系U266细胞中,蛋白印迹检测其vdac-1蛋白及髓系细胞重要转录因子C/EBPα的表达;显微镜观察细胞形态的改变;流式细胞术检测细胞表面CD45及CD33分子的表达。结果:CD45-的U266细胞转染vdac-1基因后,高水平表达vdac-1基因和蛋白,并且髓系细胞重要转录因子C/EBPα表达增加;显微镜下观察发现细胞发生类似于髓系细胞形态学改变,如胞浆突起、胞核折叠;流式细胞术检测发现转染vdac-1基因的U266细胞不表达CD45,而CD33表达增加,其中CD33阳性细胞占细胞总数的30%以上。结论:vdac-1基因能够调控MM细胞表达髓系分化抗原CD33,使细胞发生髓系细胞形态学改变,CD33有望成为MM治疗新靶点。     

四色流式分析多发性骨髓瘤细胞免疫表型及稳定性

目的:探讨多发性骨髓瘤(MM)细胞的免疫表型特征及稳定性。方法:用四色流式细胞术检测28例MM患者恶性浆细胞表面免疫表型,并对其中12例MM初诊患者在2～4m非单抗治疗后重复检测免疫表型。结果:与正常浆细胞不同,多数MM恶性浆细胞异常表达CD56(89.3%)、CD117(50.0%)、CD28(25%)、CD20(14.3%),不表达CD19(96.4%),近一半的患者不表达CD27(42.8%);恶性浆细胞表面免疫表型不稳定,12例初诊患者中有5例(41.67%)患者化疗后抗原表达有改变;CD38++CD45dim～-CD19-CD56+表型的恶性浆细胞所占圈定浆细胞百分率(NP)与骨髓形态学检测瘤细胞负荷有很强的正相关性(γ=0.675,P<0.01),并且可以区分稳定期和进展期患者(P<0.05)。结论:恶性浆细胞表面异常表达CD56,不表达CD19,部分表达CD117、CD28、CD20,近一半患者CD27表达缺失。部分初诊患者非单抗治疗后,恶性浆细胞表面抗原表达发生变化。NP可以稳定区分稳定期和进展期患者,是衡量病情的新指标。     

流式细胞技术在多发性骨髓瘤诊断中的应用研究

目的:探讨CD38/138/45/SSC及CD38/56/19/45/SSC四色组合流式细胞术在多发性骨髓瘤(MM)患者诊断中的临床应用。方法:采用多参数流式细胞技术,通过CD38/CD45双抗体设门,以CD38/138/45/SSC及CD38/56/19/45/SSC四色组合检测33例确诊MM患者骨髓中的浆细胞,并进行浆细胞胞浆κ/λ的检测。结果:33例MM患者中均可检测到CD38~(++)/CD138~+/CD45~(+/-)细胞群,在该细胞群中并未找到56~-/19~+的正常浆细胞,免疫表型为45~-/56~+/19~-标本26例,占79%,45~-/56~-/19~-标本5例,占15%,45~+/56~+/19~-标本2例,占6%。对上述CD38~(++)/138~+标本进行胞浆κ/λ检测,均为单克隆瘤细胞。结论:CD38/138/45/SSC及CD38/56/19/45/SSC四色流式细胞检测能精确发现恶性变的浆细胞,在MM诊断中有重要的临床意义和诊断价值。     

自体CIK细胞输注在老年多发性骨髓瘤维持治疗中的临床观察

目的:评估自体CIK细胞输注在老年多发性骨髓瘤(MM)维持治疗中的有效性和安全性。方法:选取5例已获得完全缓解或部分缓解的老年MM患者,采集外周血单个核细胞,在体外经干扰素-γ、白细胞介素-2、抗CD3单克隆抗体诱导成CIK细胞,分次回输至患者体内,总量为(1~3)×1010。1个月为1个疗程,连用3个疗程,再3个月为1个疗程,观察疗效和生活质量。结果:随访9~23个月,5例患者共接受了32个疗程的CIK细胞治疗,不良反应少而轻微,输注后CD3+、CD3+CD8+、CD3+CD56+细胞比例显著增高(P0.05),能有效杀伤残留肿瘤细胞,维持疾病缓解状态,提高生活质量(P0.05)。结论:自体CIK细胞输注用于本组5例老年MM患者的维持治疗安全有效。     

初诊多发性骨髓瘤患者CD56表达与以硼替佐米联合来那度胺为基础方案诱导治疗疗效的临床研究

分析初诊多发性骨髓瘤(MM)患者CD56表达差异与其预后的相关性,以硼替佐米联合来那度胺(VRd)为基础方案诱导治疗疗效的临床研究。回顾性分析山西医科大学第三医院血液科2015年1月—2021年8月初诊MM患者160例,分为CD56表达阳性组(CD56⁺)和CD56表达阴性组(CD56^-),比较2组临床特征及总生存期(OS);其中以VRd方案诱导治疗4个疗程的初诊MM患者共64例患者,在4个疗程后进行疗效评估及OS的分析,比较2组疗效评估的差别。CD56⁺组109例(68.1%),CD56^-组51例(31.9%);CD56^-组β2微球蛋白(β2-MG)及乳酸脱氢酶水平(LDH)较CD56⁺组更高(P=0.04,P=0.03);CD56^-组更易发生髓外病变(P<0.01);CD56⁺组患者OS更长(P=0.04);CD56^-、β2-MG≥8 mg/L是MM患者OS的独立预后不良因素(P&...     

多发性骨髓瘤免疫表型分析及预后

目的 :探讨多发性骨髓瘤 (MM )骨髓免疫表型的特点及外周血CD 38+ 细胞的表达。方法 :采用间接免疫荧光法对 4 1例MM患者进行免疫表型分析。结果 :①MM患者外周血及骨髓CD38+ 表达率均为 90 .2(37/ 4 1) %。②按三个不同病期分为初发期组、稳定期组及进展期组患者的骨髓细胞CD38及HLA DR阳性表达率分别为 88.9%、5 5 .6 % ;88.9%、6 6 .7% ;10 0 .0 %、4 0 .0 %。其外周血三个病期CD38+ 细胞表达率分别为88.9%、88.9%及 10 0 .0 % ,不管是外周血还是骨髓三组相比均差异无统计学意义 (P >0 .0 5 )。③按形态学分类将MM初诊患者分为成熟型和幼稚型 ,两个组CD38+ 及HLA DR阳性表达率分别为 83.3%、6 6 .7% ;10 0 .0 %、4 6 .7% ,两组相比差异无统计学意义 (P >0 .0 5 ) ;④B淋巴系统抗原在初发期、进展期中CD19、CD2 0呈部分表达 ,而在稳定期则全部消失 ;μ、κ、λ在三个组中均呈低表达 ,7例 μ、κ、λ阳性表达的病例中有 6例为骨髓瘤细胞 >6 0 %以上。结论 :CD38+ 对于诊断MM有独特意义 ,尤其结合外周血意义更大 ;B淋巴系统抗原CD19、CD2 0可能是一个预后不良因素 ,提示CD19+ 、CD2 0 + 患者预后差 ;认为骨髓中瘤细胞≥ 6 0 %对于轻链、重链的检出有价值。     

多发性骨髓瘤黏附分子CD11 a、CD49d 的表达特点及临床意义

目的研究多发性骨髓瘤(MM)患者骨髓单个核细胞黏附分子CD11a、CD49d的表达,并探讨其发病机制.方法用流式细胞仪检测CD11a、CD49d在MM组及对照组2组骨髓标本单个核细胞表达的平均荧光强度和荧光阳性细胞百分率.结果MM患者骨髓单个核细胞CD49d的表达荧光强度较对照组增强(P＜0.05);CD11a的表达阳性细胞百分率及荧光强度均较对照组增强(P＜0.05,P＜0.01);进展期表达较平台期明显增强(P＜0.05,P＜0.01).结论MM单个核细胞表面CD49d的表达增强,导致患者骨髓细胞与细胞及细胞与骨髓基质间的黏附异常,可能参与了MM的病理过程,而CD11a在MM的平稳期表达较低,进展期表达明显增强,表明CD11a在MM增殖中起作用.     

CD4~+ CD25~(high) CD127~(low/-)调节性T细胞在多发性骨髓瘤患者外周血的表达

目的探讨多发性骨髓瘤(MM)患者外周血CD4+CD25highCD127low/-调节性T(Tregs)细胞表达水平及临床意义。方法采用流式细胞术(FCM)检测30例MM患者和20例健康对照者外周静脉血CD4+CD25highCD127low/-Tregs细胞水平。结果 1 MM组CD4+CD25high CD127low/-Tregs细胞表达明显高于正常对照组(P<0.05)。2MM组不同国际分期系统(ISS)分期CD4+CD25highCD127low/-Treg细胞水平均高于正常对照组,随着疾病进展,Ⅲ期明显高于Ⅱ期(P<0.05)。3MM组化疗后外周血CD4+CD25highCD127low/-Tregs细胞水平较化疗前显著下降(P<0.05)。结论 MM患者外周血CD4+CD25highCD127low/-Treg细胞比例明显升高,可能是MM免疫逃逸的一个重要机制;进展期Tregs的变化为MM患者及时进行临床干预提供实验依据。     

《中国多发性骨髓瘤诊治指南》2022年修订新诊断患者治疗部分解读

《中国多发性骨髓瘤诊治指南》2022年修订对于新诊断多发性骨髓瘤（multiple myeloma,MM）患者的治疗选择有了一定的更新,本文就新诊断MM患者的治疗部分做如下解读。     

真实世界伊沙佐米治疗多发性骨髓瘤疗效与安全性研究

目的 评估真实世界伊沙佐米治疗多发性骨髓瘤（MM）的疗效和安全性。方法 回顾性分析2019年1月至2021年1月来自中国贫血东部协作组单位（无锡市人民医院等）伊沙佐米治疗72例MM患者的血液学、细胞遗传学及疗效、安全性等数据。结果 难治/复发MM(RRMM)患者接受伊沙佐米的中位治疗周期为6.0(3.0,7.0)个，总体有效率（ORR）为56.5%；新诊断MM(NDMM)患者中位治疗周期为4.5(4.0,9.5)个，总体有效率为85.8%；转换维持治疗组患者中位治疗周期为5.0(3.0,8.0)个，其中25.8%的患者缓解程度加深；不良事件（AEs）的总体发生率为26.5%。结论在真实世界中，伊沙佐米对RRMM、NDMM或是维持转换MM患者，具有良好的疗效和安全性。     

血清乳酸脱氢酶水平与初诊老年多发性骨髓瘤病人预后的相关性分析

目的探讨血清乳酸脱氢酶(LDH)与初诊老年多发性骨髓瘤(MM)临床指标的相关性及其预后意义。方法回顾性分析我院2009年7月至2016年1月年龄≥65岁的78例初诊老年MM病人的临床资料,并绘制病人的Kaplan-Meier生存曲线,分析初诊时血清LDH水平与病人预后的关系。根据病人的LDH水平分为LDH正常组和LDH升高组,并根据荧光原位杂交技术检测结果对60例资料完整的MM病人进行修正的国际分期系统(R-ISS)分期,比较其与国际分期系统(ISS)分期对预后判断的准确性。结果所有病人中位随访时间为16.5个月,初诊时LDH水平升高者占11.5%(9/78)。LDH正常组中位生存期(OS)和中位无进展生存期(PFS)分别为44.0个月、23.0个月,LDH升高组分别为14.0个月、12.0个月,2组间比较,差异有统计学意义(P<0.01)。COX多因素回归分析显示,LDH水平升高是老年MM病人OS的独立不良预后因素(HR=5.998,95%CI2.454~14.664,P<0.001)。另外,ISS分期Ⅱ期和Ⅲ期病人的中位OS差异无统计学意义(44.0个月比39.0个月,P=0.713),中位PFS差异也无统计学意义(26.0个月比20.0个月,P=0.569);而R-ISS分期Ⅱ期和Ⅲ期病人的中位OS差异有统计学意义(44.0个月比15.5个月,P<0.001),中位PFS差异无统计学意义(21.0个月比14.0个月,P=0.097)。结论LDH水平是判断老年MM病人预后的重要指标,基于其基础上的R-ISS分期在预后判断中要优于ISS分期。     

Outcomes of young adults (aged ≤ 40 years) with newly diagnosed multiple myeloma after up-front autologous stem cell transplant

Multiple myeloma (MM) primarily affects older patients. There are scarce data on the outcomes of young adults undergoing autologous transplantation (auto-HCT). In this single-centre analysis, we included 117 younger patients, with a median age of 37 years (range 22–40) at transplant. Seventeen (15%) patients had high-risk cytogenetics. Before transplant, 10% of patients achieved ≥CR and 44% achieved ≥VGPR. At best post-transplant response, 56% and 77% of patients achieved ≥CR and ≥VGPR respectively. With a median follow-up for survivors of 72.6 months (range 0.9–238.0), median PFS and OS were 43.1 months (95% CI 31.2–65.0) and 146.6 months (95% CI 100.0–208.1) respectively. Patients who underwent auto-HCT after 2010 had better median PFS (84.9 months vs. 28.2 months, p < 0.001) and OS (NR vs. 91.8 months, p < 0.001) compared with those transplanted earlier. In multi-variate analysis, achieving ≥CR as best post-transplant response was associated with improved PFS (HR [95% CI] 0.55 [0.32–0.95], p = 0.032), while achieving ≥VGPR was predictive of superior OS (0.32 [0.16–0.62], p < 0.001). Three patients (3%) developed a second primary malignancy. Younger MM patients had durable survival after auto-HCT, which further improved after the availability of novel anti-myeloma drugs in recent years. Depth of response following transplant remains a key predictor of survival.     

Multiparameter flow cytometric remission is the most relevant prognostic factor for multiple myeloma patients who undergo autologous stem cell transplantation

Minimal residual disease (MRD) assessment is standard in many hematologic malignancies but is considered investigational in multiple myeloma (MM). We report a prospective analysis of the prognostic importance of MRD detection by multiparameter flow cytometry (MFC) in 295 newly diagnosed MM patients uniformly treated in the GEM2000 protocol VBMCP/VBAD induction plus autologous stem cell transplantation [ASCT]). MRD status by MFC was determined at day 100 after ASCT. Progression-free survival (PFS; median 71 vs 37 months, P < .001) and overall survival (OS; median not reached vs 89 months, P = .002) were longer in patients who were MRD negative versus MRD positive at day 100 after ASCT. Similar prognostic differentiation was seen in 147 patients who achieved immunofixation-negative complete response after ASCT. Moreover, MRD(-) immunofixation-negative (IFx(-)) patients and MRD(-) IFx(+) patients had significantly longer PFS than MRD(+) IFx(-) patients. Multivariate analysis identified MRD status by MFC at day 100 after ASCT as the most important independent prognostic factor for PFS (HR = 3.64, P = .002) and OS (HR = 2.02, P = .02). Our findings demonstrate the clinical importance of MRD evaluation by MFC, and illustrate the need for further refinement of MM re-sponse criteria. This trial is registered at http://clinicaltrials.gov under identifier NCT00560053.     

Prognostic factors for survival after autologous transplantation: a single centre experience in 133 multiple myeloma patients

Autologous stem cell transplantation (ASCT) has an established role in the treatment of symptomatic multiple myeloma (MM). Our aim was to analyse the impact of selected prognostic parameters on the survival of patients with MM after ASCT. The new International Staging System (ISS) was also evaluated. A total of 133 MM patients were transplanted in our centre between 1995 and 2002. Following ASCT, 35% of patients were in complete remission (CR) and 60% were in partial remission (PR). The median progression-free (PFS) and overall (OS) survival from transplantation were 29.5 and 68.8 months, respectively. Transplant-related mortality (TRM) was 3%. On multivariate analysis, factors associated with significantly shorter OS were lack of CR after transplant (P = 0.002, hazard ratio (HR): 3.1), stage 3 according to ISS (P = 0.001, HR: 3.0) and age at transplant over 60 years (P = 0.035, HR: 2.0). The status of disease before ASCT did not significantly affect PFS and OS after transplantation. We conclude that ASCT is a safe and effective procedure in MM patients, associated with low TRM. The survival after ASCT was dependent on response after ASCT, stage according to ISS and age.     

Outcome after autologous and allogeneic stem cell transplantation for patients with multiple myeloma: impact of graft-versus-myeloma effect

A total of 228 patients with multiple myeloma (MM), 166 patients receiving autologous transplantation (124 PBSC and 38 BM) and 66 patients receiving T-cell-depleted allogeneic transplantation were analyzed to compare overall survival (OS), progression-free survival (PFS) and risk of relapse. Patients receiving autologous transplantation had a significantly improved OS (P=0.006) and PFS (P=0.002) at 2 years with OS and PFS for autologous transplant 74% and 48%, respectively, compared with 51% and 28% for allogeneic transplantation. By 4 years after transplantation, outcome was similar with OS and PFS for autologous transplantation 41% and 23%, respectively, compared with 39% and 18% for allogeneic transplantation. The 4-year cumulative incidence of nonrelapse mortality was significantly higher in patients receiving allogeneic transplantation (24% vs 13%) (P=0.004). Relapse was the principle cause of treatment failure for both groups; however, there was a significantly reduced risk of relapse associated with allogeneic transplantation at 4 years: 46% for allograft vs 56% for autograft (P=0.02). Despite a lower risk of relapse after allogeneic transplantation, autologous transplantation is associated with improved OS and PFS compared with allogeneic transplantation in patients with MM. Strategies focused on reducing nonrelapse mortality in allogeneic transplantation may translate into an improved outcome for patients receiving allogeneic transplantation.     

免疫治疗相关性甲状腺功能异常与不可切除/晚期肝细胞癌患者预后改善相关

摘要 目的：探讨免疫治疗相关性甲状腺功能异常与不可切除/晚期肝细胞癌（HCC）患者预后改善的相关性。方法：回顾性分析45例接受免疫检查点抑制剂（ICIs）治疗的不可切除/晚期HCC患者。根据ICIs治疗过程中是否出现免疫相关性甲状腺功能异常分为甲状腺功能正常组（28例）和异常组（17例）,比较2组患者的预后和免疫应答情况,主要终点指标为中位总生存期（OS）、无进展生存期（PFS）,次要终点指标为疾病控制率（DCR）。结果：所有患者的中位OS、PFS分别为10.8个月（95% CI ：3.0~18.6）和5.0个月（95% CI ：3.0~12.2）。正常组中位OS为5.8个月（95% CI ：3.7~7.9）,异常组中位OS尚未达到（ P =0.026）。异常组中位PFS长于正常组（8.2个月 vs. 3.1个月, P =0.011）,DCR高于正常组（52.9% vs. 21.5%,P =0.030）。多因素Cox回归分析显示,甲状腺功能异常是达到6个月OS（ HR=0.213,95%CI ：0.048~0.944, P =0.042）和PFS（ HR=0.383,95%CI：0.151~0.967,P =0.042）的独立影响因素;甲状腺功能异常（ HR=0.403 ,95%CI：0.185~0.877,P =0.022）、基线无大血管侵犯（MVI）（ HR=2.848,95%CI：1.406~5.768,P =0.004）、Child-Pugh A级（ HR=2.404,95%：1.099~5.255,P =0.028）与12个月PFS相关。结论：免疫治疗相关性甲状腺功能异常的不可切除/晚期HCC患者预期生存和免疫应答效果更佳。治疗期间出现甲状腺功能异常、基线无MVI、Child-Pugh A级与患者预后改善相关。     

Cetuximab plus FOLFOX6 or FOLFIRI in metastatic colorectal cancer: CECOG trial

AIM: To investigate efficacy and safety of cetuximab combined with two chemotherapy regimens in patients with unresectable metastatic colorectal cancer (mCRC). METHODS: Randomized patients received cetuximab with 5-fluorouracil (5-FU), folinic acid (FA) and oxaliplatin (FOLFOX) 6 (arm A, n = 74) or 5-FU, FA and irinotecan (FOLFIRI) (arm B, n = 77). KRAS mutation status was determined retrospectively in a subset of tumors (n = 117). RESULTS: No significant difference was found between treatment arms A and B ...     

Predictors of prolonged survival after allogeneic hematopoietic stem cell transplantation for multiple myeloma

A total of 149 patients with multiple myeloma (MM) who received allogeneic hematopoietic stem cell transplantation (allo-HCT) with myeloablative (MAC; n = 38) or reduced-intensity conditioning (RIC; n = 110) regimens at MD Anderson Cancer Center were evaluated. Of the total, 120 (81%) patients had relapsed or had refractory disease. Median age of MM patients was 50 (28-70) years with a followup time of 28.5 (3-164) months. The 100-day and 5-year treatment related mortality (TRM) rates were 17% and 47%, respectively. TRM was significantly lower with RIC regimens (13%) vs. 29% for MAC at 100 days (P = 0.012). The cumulative incidence of Grade II-IV acute graft-versus-host disease (GVHD) was 35% and chronic GVHD was 46%. PFS and OS at 5 years were 15% and 21%, respectively. In multivariate analysis, allo-HCT for primary remission consolidation was associated with longer PFS (HR 0.35; 95% CI, 0.18-0.67) and OS (HR 0.29; 95% CI 0.15-0.55), while absence of high-risk cytogenetics was associated with longer PFS only (HR 0.59; 95% CI 0.37-0.95). We observe that TRM has decreased with the use of RIC regimens, and long-term disease control can be expected in a subset of MM patients undergoing allo-HCT. Further studies should be conducted in carefully designed clinical trials in this patient population.     

Clinicopathological and prognostic characteristics of CD56-negative multiple myeloma

We analysed CD56 expression in 70 patients with multiple myeloma (MM) to determine its clinicopathological and prognostic significance. Fifty-five (79%) patients were CD56+. CD56- patients (n = 15) had higher beta2 microglobulin levels and a higher incidence of extramedullary disease, Bence Jones protein, renal insufficiency and thrombocytopenia than CD56+ patients. Their myelomas more frequently had a plasmablastic morphology. Overall survival was significantly lower in CD56- than CD56+ patients (22 vs 63 months, P = 0.0002). We conclude that CD56- MM is a discrete entity associated with more aggressive disease. The higher incidence of plasmablastic cases suggested that CD56- MM may develop from a less mature plasma cell than CD56+ MM.     

Phosphatase and tensin homolog expression related to cetuximab effects in colorectal cancer patients: a meta-analysis

AIM: To investigate the correlation between expression of phosphatase and tensin homolog (PTEN) and cetuximab effects in colorectal cancer.METHODS: We searched PubMed, EMBASE and ASCO to identify eligible studies. Finally, 8 randomized control studies were included in the meta-analysis. STATA 10.0 Software was used to investigate heterogeneity among individual studies and to summarize all the studies. Risk ratios (RRs) and hazard ratios (HRs) with 95% confidence intervals (CIs) were used to assess the strength of the association.RESULTS: Compared with 20 of 266 patients with loss of PTEN, 206 of 496 patients with intact PTEN protein expression had a better objective response rate to cetuximab-based therapy (RR, 4.75; 95% CI, 2.59-8.72; P < 0.001). PTEN positivity was associated with better progression-free survival (PFS) (HR, 0.675; 95% CI, 0.473-0.964; P = 0.031) but not with better overall survival (OS) (HR, 0.608; 95% CI, 0.411-0.899; P = 0.013). In patients with KRAS wild-type status, PTEN positivity did not predict a longer PFS or OS (PFS: HR, 0.707; 95% CI, 0.440-1.138; P = 0.154; OS: HR, 0.943; 95% CI, 0.646-1.377; P = 0.761).CONCLUSION: Expression of PTEN is related to the effect of cetuximab in colorectal cancer patients and should be considered in treatment with cetuximab.