难治复发急性髓系白血病患者十种常见突变基因的突变组分析

目的 探讨难治复发急性髓系白血病(AML)患者中常见的10种突变基因发生的规律.方法 选取难治复发AML患者148例.基因测序检测并分析患者骨髓样本中10种常见突变基因,包括激酶类基因FLT3和KIT,转录因子基因CEBPA、NPM1和PHF6,以及表观遗传类基因ASXL1、DNMT3A、IDH1、IDH2和TET2.结果 在62.16％(92/148)的患者中检测到上述基因突变阳性,其中10.14％(15/148)的患者同时携带多个基因的突变.FLT3-ITD突变率最高(19.59％,29/148),其次为KIT(12.84％,19/148)和CEBPA(11.49％,17/148)突变.KIT突变常单独出现,而IDH1/2突变常伴随其他基因突变.激酶类基因FLT3和KIT突变互斥,转录因子基因和表观遗传基因也存在同类互斥现象.以35岁为界限分组,≤35岁的患者多携带单个基因的突变[61.77％(63/102)比31.11％(14/45),P＜ 0.05],而＞35岁的患者多个基因突变(≥2个)的携带率高[20.00％(9/45)比4.90％(5/102),P＜ 0.05].＞35岁患者组中NPM1突变率高于≤35岁组[20.00％(9/45)比2.94％(3/102),P＜ 0.05].结论 研究发现难治复发AML患者中常见基因突变的组合具有一定的规律,与基因的功能分类和患者的年龄有关.     

DNMT3A、FLT3-ITD合并NPM1基因突变正常核型急性髓系白血病患者的临床特征及预后分析

目的 分析DNMT3A、FLT3-ITD合并NPM1基因突变的正常核型急性髓系白血病(AML)患者的临床特征,探讨该类患者的预后.方法 回顾性分析2005年12月至2014年6月就诊的109例正常核型AML患者的临床特征及预后.其中DNMT3A/FLT3-ITD/NPM1+AML患者共25例,单独FLT3-ITD+ AML患者共32例,单独NPM1+AML患者24例,单独DNMT3A+ AML患者28例.结果 25例DNMT3A/FLT3-ITD/NPM1+ AML患者平均年龄46岁,伴有高白细胞(81.7×109/L)及较高的骨髓原始细胞(66.3％).其中,17例选择大剂量化疗,8例行异基因造血干细胞移植,与DNMT3A+、FLT3-ITD+、NPM1+AML组比较,差异均无统计学意义.25例DNMT3A/FLT3-ITD/NPM1+患者的3年总生存率为17.65％,3年无病生存率为13.88％,与DNMT3A+、FLT3-ITD+、NPM1+AML组差异均有统计学意义(P值分别为0.049 9、0.036 1、0.013 4),3年无病生存率差异无统计学意义(均P＞ 0.05).结论 DNMT3A、FLT3-ITD合并NPM1基因突变的正常AML患者往往合并高的外周血白细胞及骨髓原始细胞,预后较差.     

TET2合并DNMT3A突变及其他共存基因突变对急性髓系白血病患者预后的影响

  目的  探究TET2合并DNMT3A突变及其他共存基因突变对成人非M3型急性髓系白血病（acute myeloid leukemia,AML）患者预后的影响。  方法  回顾性分析2018年1月至2021年9月于南昌大学第一附属医院确诊的初治且行血液肿瘤相关突变基因外显子二代测序检测的512例成人 AML（非 M3 型）患者的临床资料,分析患者的临床特征、疗效及生存情况。  结果  本研究共纳入110例AML患者,TET2突变组64例,DNMT3A单突变组46例。男性50例（45.5%）,中位年龄54（15～79）岁。TET2基因突变频率为12.5%（64/512）,98.4%（63/64）患者突变基因数≥2个,每例患者平均合并5.2个突变基因。NPM1（43.8%）、DNMT3A（42.2%）、FLT3-ITD/TKD（40.6%）、CEBPA（26.6%）、TTN（20.3%）为TET2突变常见的共存突变基因。TET2合并DNMT3A突变患者初次诱导完全缓解（complete response,CR）率为46.2%,略低于TET2单突变患者的76.9%（P=0.077）,与DNMT3A单突变患者无显著性差异（P=0.952）。TET2合并DNMT3A突变患者的中位总生存（median overall survival,mOS）时间为9.5个月,明显低于TET2单突变患者（P=0.002）,而与DNMT3A单突变患者无显著性差异（P=0.414）。三者的中位无复发生存（median relapse- free survival,mRFS）时间无显著性差异（P>0.05）。在TET2突变背景下,合并K/NRAS突变患者的CR率为28.6%,明显低于无K/NRAS突变患者的75.0%（P=0.030）,合并FLT3-ITD突变患者的mOS明显短于无FLT3-ITD突变患者（P=0.030）。多因素分析显示年龄≥60岁、合并FLT3-ITD突变、初次诱导未达CR是影响TET2突变AML患者总生存时间（overall survival,OS）的独立危险因素,DNMT3A突变不影响TET2突变患者OS。  结论  TET2突变是AML患者常见突变,且常合并共存基因突变。共存基因突变与TET2突变共同影响AML患者预后。基于二代测序的基因突变检测对指导AML精确分层及精准治疗具有重要意义。      

伴TET2基因突变急性髓系白血病成年患者临床特征及突变对疗效和预后的影响

目的:探讨伴TET2基因突变急性髓系白血病（AML）成年患者的临床特征以及突变对疗效和预后的影响。方法:选择2017年3月至2021年4月于济宁市第一人民医院就诊的123例初诊AML成年患者（除外急性早幼粒细胞白血病）,采用二代测序方法检测包括TET2突变在内的24种AML相关基因突变情况。根据有无TET2基因突变将患者分为TET2基因突变型组及TET2基因野生型组,比较两组患者临床病理特征及近期疗效和生存差异。结果:123例患者中,28例（22.8%）检测到TET2突变。与TET2基因野生型组患者相比,TET2基因突变型组患者的年龄更高[（59±15）岁比（49±16）岁, t=2.984, P=0.003],更易出现法、美、英（FAB）协作组分型的M 4、M 5型[75.0%（21/28）比51.6%（49/95）, χ2=4.838, P=0.028],AML患者CD34阳性率更低[46.4%（13/28）比72.6%（69/95）, χ2=6.685, P=0.010];TET2基因突变更易伴发ZRSR2突变[10.7%（3/28）比1.1%（1/95）, P=0.037]和NPM1突变[35.7%（10/28）比17.9%（17/95）, χ2=4.008, P=0.045],而较少伴发IDH1/2基因突变[0比17.9%（17/95）, P=0.012]。两组间性别、初诊时外周血白细胞计数、血红蛋白水平、血小板计数、骨髓原始细胞比例、细胞遗传学及欧洲白血病网络（ELN）危险度分层方面差异均无统计学意义（均 P>0.05）。两组患者1个疗程化疗和去甲基化治疗的总有效率（ORR）差异均无统计学意义[75.0%（12/16）比66.7%（42/63）, χ2=0.410, P=0.522;66.7%（4/6）比44.4%（8/18）, P=0.640]。TET2基因突变型组和野生型组总生存（OS）差异无统计学意义[中位OS时间：23个月（95% CI 5~41个月）比35个月（95% CI 18~52个月）, P=0.498]。 结论:在AML成年患者中,TET2基因突变与高龄、M 4和M 5亚型、AML细胞低表达CD34相关。TET2基因突变易伴随ZRSR2、NPM1基因突变,而不易伴随IDH1或IDH2基因突变。TET2基因突变对未进行危险度分层AML患者的总体疗效和生存可能无显著影响。     

急性髓系白血病相关基因差异表达分析并文献复习

目的 探讨急性髓系白血病（AML）相关基因表达水平在白血病发生过程中的作用,进一步阐明AML可能的发病机制.方法 采用高通量测序技术对1例急性髓系白血病部分分化型（AML-M2）患者初诊时及缓解后外周血样本进行转录组测序,统计表达基因及差异表达基因,并对差异表达基因进行基因本体论（GO）和基因所在的代谢通路（KEGG pathway）富集分析,对白血病相关基因JAK1、JAK2、TET2、ASXL1、IDH2、WT1、BCR、ABL1、JAK3、GATA2、HOXA9、FLT3、IDH1表达情况进行筛选分析.结果 该AML-M2患者外周血中WT1、BCR、ABL1、JAK3、GATA2、HOXA9、FLT3、IDH1为差异表达基因.结论 高通量测序技术可检测疾病某一时期大部分表达基因.通过初诊时及缓解期基因表达水平分析,推断个别基因除了可以通过获得性突变致病外,还可能通过差异表达在白血病发病中发挥作用.     

骨髓增生异常综合征患者基因突变情况分析

目的 分析骨髓增生异常综合征(MDS)患者的基因突变情况.方法 选择2016年1月至2017年7月中国中医科学院西苑医院初诊的47例MDS患者,采用NGS 127-gene panel检测基因突变,分析基因突变与临床特征的关系.结果47例MDS患者中,31例(66.0％)检测到基因突变,涉及有临床意义的突变基因23个,检出率>5％的基因共7个,由高到低依次为U2AF1(23.4％)、SF3B1(12.8％)、ASXL1(10.6％)、TET2(8.5％)、BCOR(8.5％)、TP53(8.5％)、DNMT3A(6.4％).31例基因突变患者中,16例(51.6％)存在2个以上基因协同突变,其中12例患者的协同基因突变存在于不同基因功能组内,高于单一基因功能组内的基因协同突变(4例).IDH2-KRAS、IDH2-SRSF2、IDH2-STAG2、KRAS-SRSF2、KRAS-STAG2、RUNX1-PHF6、EZH2-ASXL1、EZH2-ZRSR2、NPM1-NRAS基因之间有共存关系(均P<0.05).JAK2、KRAS、NRAS、SH2B3四个信号通路相关的变异等位基因频率(VAF)较低,处于亚克隆地位.1例JAK2突变见于MDS-U患者;1例SH2B3基因突变见于核型预后极高危患者;2例SETBP1以及2例EZH2突变均见于修订的国际预后评分系统(IPSS-R)预后高危患者.结论 MDS患者常见突变基因为U2AF1、SF3B1、ASXL1、TET2.不同基因功能组内的基因倾向于协同突变.基因突变可用于判断MDS患者预后,作为其治疗的靶点.     

急性髓细胞白血病NPM1和FLT3-ITD突变检测临床意义分析

目的:研究核仁磷酸蛋白1(NPM1)FMS样酪氨酸激酶3(FLT3)基因内部串联重复(ITD)突变在急性髓细胞白血病(AML)中发生的情况,并了解其临床特征及预后意义.方法:分别应用高分辨熔解曲线(HRM)和变性高效液相色谱技术(DHPLC)检测103例初诊AML患者NPM1和FLT3-ITD突变情况,并结合临床资料进行分析.结果:103例初诊AML患者中,31例发现NPM1基因突变,20例发现FLT3-ITD突变,阳性率分别为30.1％和19.4％,而在核型正常组中所占的比例分别为47.6％(20/42)和26.2％(11/42).FLT3-ITD突变型外周血白细胞计数高,t=2.21,P=0.037;骨髓原始细胞比例高,t=2.44,P=0.023;NPM1突变型亦表现为高外周血白细胞计数,t=2.24,P=0.034.在非M3患者中,NPM1突变型的CR与野生型差异无统计学意义,但第1次化疗的CR(76.9％)明显高于野生型患者(35.0％),x2=12.78,P=0.000 35;FLT3-ITD突变型患者的CR为58.8％,野生型患者为82.6％,两者比较,x2=4.48,P=0.034;FLT3-ITD突变型患者第1次化疗的CR为17.6％,而野生型患者为55.1％,两者比较,x2=6.23,P=0.012.31例NPM1基因突变中有6例合并FLT3-ITD突变,NPM1+/FLT3-ITD-组的CR最高(85.0％),1年内RR率最低(17.6％);NPM1-/FLT3-ITD+组的CR最低(54.5％),1年内RR率最高(50.0％).结论:NPM1和FLT3-ITD突变是AML患者常见的分子遗传学异常,与预后密切相关,可成为目前细胞遗传学预后分组的重要补充,对于指导AML患者的个体化治疗具有重要的临床价值.     

正常核型急性髓系白血病中具有预后意义的基因改变

 急性髓系白血病（AML）是一类具有遗传学异质性的疾病,大约40 %～49 ％的成年人AML及25 %的儿童AML表现为正常核型,并被归入中危组,但因获得性基因改变,如FLT3突变、NPM突变、CEBPA 突变、MLL突变、KIT突变以及BAALC、MN1、ERG1 及EVI1基因过表达等,导致了该组患者预后的显著异质性,现就几种常见的分子遗传学异常作简要的综述,讨论其在正常核型AML中预后意义及临床价值。     

急性髓系白血病全外显子组测序基因突变特点及其与疗效的关系

目的探讨急性髓系白血病(AML)患者全外显子组测序(WES)基因突变特点及其与疗效的关系。方法回顾性分析2014年12月至2019年9月安徽医科大学第二附属医院收治的30例AML患者资料, 总结WES结果及疾病类型、疾病分型、遗传学预后分层、疗效等情况, 比较不同临床特征和遗传学预后分层患者突变类型及突变频率。结果 30例AML患者中26例(86.7%)至少发生1个基因突变, 其中突变频率>10.0%的基因依次为NRAS、RUNX1、TET2、CEBPA、IDH2、ASXL1, 这些突变基因的功能涉及信号通路、转录因子、表观遗传、RNA剪接及其他生物学功能, 19例(63.3%)在突变方式上呈现为多种组合的联合突变。在年龄、性别、疾病类型、疾病分型及遗传学预后分层各组中突变率差异均无统计学意义(均P>0.05)。融合基因阳性8例中突变7例, 平均突变频率175.0%(14/8);融合基因阴性22例中突变19例, 平均突变频率213.6%(47/22);两组间突变频率差异有统计学意义(P=0.001)。持续缓解14例中突变11例, 平均突变频率157.1%(22/14);复发...     

急性髓系白血病患者骨髓涂片FMS样酪氨酸激酶3、NPM1和c-kit基因检测及临床研究

目的：探讨提取贮存骨髓涂片DNA的方法,通过对急性髓系白血病（AML）患者FMS样酪氨酸激酶3（FLT3）、NPM1及c-kit基因突变进行检测,分析三种基因突变与AML临床特征之间的关系。方法收集55例AML患者骨髓涂片,采用聚合酶链反应（PCR）、DNA测序和分子克隆方法对FLT3-内部串联重复（ITD）、NPM1和c-kit基因突变进行检测及分析,记录患者疾病缓解、进展及生存时间。结果实验证实对于低温冻存、未经瑞特染色、未用化学方法固定的骨髓涂片标本及室温贮存、经瑞特染色脱色后的标本均能用苯酚∶氯仿∶异戊醇法成功提取DNA。从骨髓涂片中提取的DNA可用于PCR、直接测序和分子克隆测序分析。在55例AML患者中,FLT3-ITD阳性10例（18.2%）,其中9例为杂合型突变,1例为纯合型突变。FLT3-ITD阳性组较阴性组完全缓解（CR）率低,无事件生存（EFS）和总生存（OS）时间短（P＜0.05）。NPM1基因杂合型突变9例（16.4%）,全部为A型突变。10个月以内NPM1突变组患者EFS率比野生组高（P＜0.05）,19个月以内NPM1突变组OS率比野生组高（P＜0.05）。9例NPM1突变患者中FLT3-ITD阳性3例,CR率由高到低依次为NPM1+ FLT3-ITD-、NPM1- FLT3-ITD-、NPM1- FLT3-ITD+、NPM1+ FLT3-ITD+（P＜0.05）,且NPM1- FLT3-ITD+是影响OS的危险因素（RR＝1.250,P＝0.005）。55例患者中,c-kit基因突变2例（3.6%）,分别为D816H突变型和D816V突变型;c-kit基因突变患者与FLT3-ITD阳性及NPM1突变患者无重叠。结论 FLT3-ITD突变为AML患者预后不良的分子标志,NPM1基因突变可能提示预后较好,NPM1- FLT3-ITD+是影响OS的危险因素。AML中c-kit基因突变率低,未见其与FLT3、NPM1基因突变重叠。     

Adverse impact of IDH1 and IDH2 mutations in primary AML: experience of the Spanish CETLAM group

The study of genetic lesions in AML cells is helpful to define the prognosis of patients with this disease. This study analyzed the frequency and clinical impact of recently described gene alterations, isocitrate dehydrogenase 1 (IDH1) and isocitrate dehydrogenase 2 (IDH2) mutations, in a series of homogeneously treated patients with primary (de novo) AML. Two-hundred and seventy-five patients enrolled in the CETLAM 2003 protocol were analyzed. IDH1 and IDH2 mutations were investigated by well-established melting curve-analysis and direct sequencing (R140 IDH2 mutations). To establish the percentage of the mutated allele a pyrosequencing method was used. Patients were also studied for NPM, FLT3, MLL, CEBPA, TET2 and WT1 mutations. IDH1 or IDH2 mutations were identified in 23.3% AML cases and in 22.5% of those with a normal karyotype. In this latter group, mutations were associated with short overall survival. This adverse effect was even more evident in patients with the NPM or CEBPA mutated/FLT3 wt genotype. In all the cases analyzed, the normal allele was detected, suggesting that both mutations act as dominant oncogenes. No adverse clinical impact was observed in cases with TET2 mutations. IDH1 and IDH2 mutations are common genetic alterations in normal karyotype AML. Favourable genotype NPM or CEBPA mutated/FLT3 wt can be further categorized according to the IDH1 and IDH2 mutational status.     

Clinical implications of novel mutations in epigenetic modifiers in AML

The studies highlighted in this article suggest that mutations in TET2 mutations may impart adverse outcome in patients with CN-AML, whereas mutations in DNMT3a may have adverse implications in a broader set of patients with AML. The data with IDH enzyme mutations are less clear, in that individual IDH1 and IDH2 mutations may have different clinical effects and the data so far have not suggested a uniform effect on outcome. Despite the exciting data indicating that mutational testing for these alterations may be clinically useful, several challenges to understanding their clinical relevance remain. First, patients may simultaneously have mutations in multiple genes described in this article (FLT3, NPM1, CEBPa, DNMT3a, IDH1/2, or TET2), and in additional genes not mentioned earlier (Ras,47 PTEN,48 PHF6,49 ASXL1,15 and RUNX145). Furthermore, comprehensive sequencing studies of well-annotated, homogeneously treated patient cohorts are needed to understand the clinical implications of integrated mutational profiling in AML. An additional challenge to using mutational analysis for TET2 and DNMT3a in clinical use is identifying a means for rapid molecular testing of these mutations. This challenge may be met by the use of non–polymerase chain reaction–based methods of target enrichment, such as hybrid capture, followed by next-generation sequencing technologies. Moreover, clinical studies evaluating the biochemical consequences of mutations in some of these genes (eg, production of 2-HG in bodily fluids from patients with IDH-mutant AML or increased hydroxymethylcytosine levels in pretreatment blast DNA in patients with TET2/IDH mutant AML) may also prove to be useful in identifying biomarkers. Alternatively, protein-based technologies such as immunohistochemistry or mass spectrometry may be used in the clinical setting to detect the mutant proteins or loss of expression of specific proteins in patients with mutations. An additional area of importance highlighted by these discoveries is the increasing realization that several of these genes encode enzymes or result in alterations in enzymatic activities, which may represent novel, tractable therapeutic targets for patients with AML. This finding may hopefully lead to the development of novel targeted therapeutics for patients with specific genetic alterations in AML. This development may be occurring now with the advent of DOT1L-targeted therapy for leukemic cells with translocations involving MLL1.50,51 Studies to identify whether the neomorphic enzymatic activity of IDH1/2 mutations may be targetable or if the downstream effects of TET2 mutations can be targeted are ongoing and may lead to the development of rational epigenetic therapies that improve outcomes for patients with AML.     

Molecular Evaluation of DNMT3A and IDH1/2 Gene Mutation:Frequency,Distribution Pattern and Associations with Additional Molecular Markers in Normal Karyotype Indian Acute Myeloid Leukemia Patients

Mutations in the DNMT3A and IDH genes represent the most common genetic alteration after FLT3/NPM1in acute myeloid leukemia (AML). We here analyzed the frequency and distribution pattern of DNMT3A andIDH mutations and their associations with other molecular markers in normal karyotype AML patients. Fortyfivepatients were screened for mutations in DNMT3A (R882), IDH1 (R132) and IDH2 (R140 and R172) genesby direct sequencing. Of the 45 patients screened, DNMT3A and IDH mutations were observed in 6 (13.3%)and 7 (15.4%), respectively. Patients with isolated DNMT3A mutations were seen in 4 cases (9%), isolatedIDH mutations in 5 (11.1%), while interestingly, two cases showed both DNMT3A and IDH mutations (4.3%).Nucleotide sequencing of DNMT3A revealed missense mutations (R882H and R882C), while that of IDH revealedR172K, R140Q, R132H and R132S. Both DNMT3A and IDH mutations were observed only in adults, with ahigher frequency in males. DNMT3A and IDH mutations were significantly associated with NPM1, while trendstowards higher coexistence with FLT3 mutations were observed. This is the first study to evaluate DNMT3A/IDH mutations in Indian patients. Significant associations among the various molecular markers was observed,that highlights cooperation between them and possible roles in improved risk stratification.     

Landscape of TET2 mutations in acute myeloid leukemia

We investigated ten-eleven translocation 2 (TET2) mutations in acute myeloid leukemia (AML), their correlation with other gene mutations and prognostic value. By deep-sequencing, 131 somatic TET2 mutations were identified in 87/318 (27.4%) patients. Of 87 mutated cases, 44 (50.6%) carried two mutations. TET2 mutations were concomitantly observed with mutations in NPM1, FLT3-ITD, FLT3-TKD, JAK2, RUNX1, CEBPA, CBL and KRAS. However, TET2 mutations rarely concomitantly occurred with IDH1mut or IDH2mut (2/251 or 0/184; P=0.046 and P=0.003, respectively). TET2 mutations were associated with normal karyotype AML (CN-AML) (62/206 (30.1%) CN-AML vs 20/107 (18.7%) aberrant karyotype; P=0.031), higher white blood cell count (mean 65.3 vs 40.3 × 10(9)/l, P=0.023), lower platelet count (mean 68.6 vs 92.4 × 10(9)/l, P=0.03) and higher age (67.5 vs 65.2 years, P<0.001). Survival analyses were restricted to de novo CN-AML patients (n=165) and showed inferior event-free survival (EFS) of TET2 mutations compared with TET2wt (median: 6.7 vs 18.7 months, P=0.009). This negative effect of TET2 mutation on EFS was particularly observed in patients 65 years (median: 8.9 months vs not reached (n.r.), P=0.027) as well as in patients of the European LeukemiaNet favorable-risk subgroup, that is, patients harboring mutated CEBPA and/or mutated NPM1 without FLT3-ITD (median: 10.3 vs 41.3 months, P=0.048). These data support a role for TET2 as an important prognostic biomarker in AML.     

Recent Discoveries in Molecular Characterization of Acute Myeloid Leukemia

Acute myeloid leukemia (AML) is a clinically heterogeneous disease, yet it is one of the most molecularly well-characterized cancers. Risk stratification of patients currently involves determination of the presence of cytogenetic abnormalities in combination with molecular genetic testing in a few genes. Several new recurrent genetic molecular abnormalities have recently been identified, including TET2, ASXL1, IDH1, IDH2, DNMT3A, and PHF6. Mutational analyses have identified that patients with DNMT3A or NPM1 mutations or MLL translocation have improved overall survival with high-dose chemotherapy. Mutational profiling can refine prognostication, particularly for patients in the intermediate-risk group or with a normal karyotype. CD25 expression status improves prognostic risk classification in AML independent of established biomarkers. Biomarkers such as 2- hydroxyglutarate in IDH1/2-mutant AML patients predict patient responses and minimal residual disease. These recent discoveries are being incorporated into our existing molecular risk stratification as well as the exploration of new therapeutics directed to these molecular targets.     

Gene Mutation Patterns in Patients with Minimally Differentiated Acute Myeloid Leukemia

Minimally differentiated acute myeloid leukemia (AML-M0) is a rare subtype of AML with poor prognosis. Although genetic alterations are increasingly reported in AML, the gene mutations have not been comprehensively studied in AML-M0. We aimed to examine a wide spectrum of gene mutations in patients with AML-M0 to determine their clinical relevance. Twenty gene mutations including class I, class II, class III of epigenetic regulators (IDH1, IDH2, TET2, DNMT3A, MLL-PTD, ASXL1, and EZH2), and class IV (tumor suppressor genes) were analyzed in 67 patients with AML-M0. Mutational analysis was performed with polymerase chain reaction–based assays followed by direct sequencing. The most frequent gene mutations from our data were FLT3-ITD/FLT3-TKD (28.4%), followed by mutations in IDH1/IDH2 (28.8%), RUNX1 (23.9%), N-RAS/K-RAS (12.3%), TET2 (8.2%), DNMT3A (8.1%), MLL-PTD (7.8%), and ASXL1 (6.3%). Seventy-nine percent (53/67) of patients had at least one gene mutation. Class I genes (49.3%) were the most common mutated genes, which were mutually exclusive. Class III genes of epigenetic regulators were also frequent (43.9%). In multivariate analysis, old age [hazard ratio (HR) 1.029, 95% confidence interval (CI) 1.013-1.044, P = .001) was the independent adverse factor for overall survival, and RUNX1 mutation (HR 2.326, 95% CI 0.978-5.533, P = .056) had a trend toward inferior survival. In conclusion, our study showed a high frequency of FLT3, RUNX1, and IDH mutations in AML-M0, suggesting that these mutations played a role in the pathogenesis and served as potential therapeutic targets in this rare and unfavorable subtype of AML.     

Clinical impact of gene mutations and lesions detected by SNP-array karyotyping in acute myeloid leukemia patients in the context of gemtuzumab ozogamicin treatment: Results of the ALFA-0701 trial

We recently showed that the addition of fractionated doses of gemtuzumab ozogamicin (GO) to standard chemotherapy improves clinical outcome of acute myeloid leukemia (AML) patients. In the present study, we performed mutational analysis of 11 genes (FLT3, NPM1, CEBPA, MLL, WT1, IDH1/2, RUNX1, ASXL1, TET2, DNMT3A), EVI1 overexpression screening, and 6.0 single-nucleotide polymorphism array (SNP-A) analysis in diagnostic samples of the 278 AML patients enrolled in the ALFA-0701 trial. In cytogenetically normal (CN) AML (n = 146), 38% of the patients had at least 1 SNP-A lesion and 89% of the patients had at least 1 molecular alteration. In multivariate analysis, the independent predictors of higher cumulative incidence of relapse were unfavorable karyotype (P = 0.013) and randomization in the control arm (P = 0.007) in the whole cohort, and MLL partial tandem duplications (P = 0.014) and DNMT3A mutations (P = 0.010) in CN-AML. The independent predictors of shorter overall survival (OS) were unfavorable karyotype (P < 0.001) and SNP-A lesion(s) (P = 0.001) in the whole cohort, and SNP-A lesion(s) (P = 0.006), DNMT3A mutations (P = 0.042) and randomization in the control arm (P = 0.043) in CN-AML. Interestingly, CN-AML patients benefited preferentially more from GO treatment as compared to AML patients with abnormal cytogenetics (hazard ratio for death, 0.52 versus 1.14; test for interaction, P = 0.04). Although the interaction test was not statistically significant, the OS benefit associated with GO treatment appeared also more pronounced in FLT3 internal tandem duplication positive than in negative patients.