正常核型急性髓细胞白血病中CEBPA基因伴随突变模式分析

doi:10.16150/j.1671-2870.2017.05.009

摘要/Abstract

摘要： 目的：探究CEBPA基因双端突变与单端突变2种突变模式的伴随附加基因突变分布差异,并讨论其与患者预后间的潜在相关性。方法：对152例正常核型初发急性髓细胞白血病(acute myeloid leukemia,AML)患者中,25个AML驱动基因的热点区域进行二代深度测序,并结合一代测序验证检测突变位点,利用卡方检验或Fisher 检验对CEBPA基因双端突变和单端突变所伴随的附加基因突变情况进行统计分析。结果：152例正常核型的初发AML患者中,CEBPA基因双端突变43例(28.29%),CEBPA基因单端突变13例(8.55%)。携带CEBPA基因单端突变患者伴3个以上附加基因突变百分比高于CEBPA基因双端突变（P=0.043）。在携带CEBPA基因双端突变患者中,未发现伴NPM1基因突变,而在13例CEBPA基因单端突变患者中,有6例伴NPM1突变(P<0.001)。附加基因突变分布分析中,CEBPA基因单端突变患者伴更多的DNA甲基化修饰基因突变（DNMT3A、TET2、IDH1、IDH2）（P=0.046）;CEBPA基因单端突变伴随更多的叠加高危组基因突变（DNMT3A、FLT3-ITD、TP53）。结论：CEBPA单端突变比双端突变患者伴随更多的预示不良预后的附加突变,可能与CEBPA基因双端突变及单端突变2种突变模式的预后差异相关。

关键词: 急性髓细胞白血病, CEBPA伴随突变, 机制

Abstract: Objective: To investigate the different patterns of additional gene mutation accompanied with CEBPA biallelic or monoallelic mutations in patients with acute myeloid leukemia, which might be associated with different prognosis of these two kinds of mutation subtypes. Methods: A total of 152 newly diagnosed acute myeloid leukemia patients with normal karyotype were enrolled. The mutations in hot spot regions of 25 AML driver genes were screened using next-generation sequencing strategy and the discovered mutation sites were confirmed by Sanger sequencing. The difference in additional gene mutation patterns accompanied with CEBPA biallelic and monoallelic mutations were analyzed by Chi-square or Fisher's exact test. Results: In the 152 patients, CEBPA biallelic mutations were identified in 43(28.29%) cases and monoallelic mutations in 13(8.55%) cases. The ratio of more than 3 additional gene mutations in patients carried CEBPA monoallelic mutations (4 cases, 30.77%) was higher than that of the patients with CEBPA biallelic mutations (3 cases, 6.98%) (P=0.043). No NPM1 gene mutation was found in patients with CEBPA biallelic mutated, while near half of the patients (6 cases, 46.15%) had CEBPA monoallelic mutations accompanied with NPM1 mutation. CEBPA monoallelic mutated patients were more involved in genes of DNA methylation group (DNMT3A, TET2, IDH1, IDH2) than those with CEBPA biallelic mutation (7 cases, 53.85% vs 10 cases, 23.26%, P=0.046). Meanwhile, more high-risk genes (DNMT3A, FLT3-ITD, TP53) co-mutated were found in patients with CEBPA monoallelic mutations than those with CEBPA biallelic mutations. Conclusions: Compared with patients with CEBPA biallelic mutation, the CEBPA monoallelic mutation cohort have more additional gene mutations associated with adverse prognosis, and the different gene mutation patterns may lead to different prognosis. However, more large-scale investigations are needed to verify their clinical significance.

Key words: Acute myeloid leukemia, CEBPA, Additional gene mutations, Mechanism

中图分类号:

R733.7

王舒, 张赟翔, 眭竫旎, 鲁静, 范惠咏, 王超, 陈冰. 正常核型急性髓细胞白血病中CEBPA基因伴随突变模式分析[J]. 诊断学理论与实践, 2017, 16(05): 498-503.

WANG Shu, ZHANG Yunxiang, SUI Jingni, LU Jing, FAN Huiyong, WANG Chao, CHEN Bing.. Analysis of additional mutation pattern accompanied with CEBPA mutations in patients with the cytogenetically normal acute myeloid leukemia[J]. Journal of Diagnostics Concepts & Practice, 2017, 16(05): 498-503.

参考文献

[1] Szczepański T, Harrison CJ, van Dongen JJ. Genetic aberrations in paediatric acute leukaemias and implications for management of patients[J]. Lancet Oncol,2010, 11(9):880-889.
[2] Ley TJ, Miller C, Ding L, et al.Genomic and epigenomic landscapes of adult de novo acute myeloid leukemia[J]. N Engl J Med,2013,368(22):2059-2074.
[3] Papaemmanuil E, Gerstung M, Bullinger L, et al.Genomic Classification and Prognosis in Acute Myeloid Leukemia[J]. N Engl J Med,2016,374(23):2209-2221.
[4] NCCN Cliinical Practice Guidelines in Oncology-----Acute Myeloid Leukemia[R/OL]. https://www.nccn.org/National Comprehensive Cancer Network,NCCN Cliinical Practice Guidelines in Oncology-----Acute Myeloid Leukemia[R/OL]. https://www.nccn.org/National Comprehensive Cancer Network,Inc,2017,2:24.
[5] Döhner H, Estey E, Grimwade D, et al. Diagnosis and management of AML in adults:2017 ELN recommendations from an international expert panel[J]. Blood,2017 Jan 26,129(4):424-447.
[6] Ley TJ, Ding L, Walter MJ, et al.DNMT3A mutations in acute myeloid leukemia[J]. N Engl J Med,2010,363(25):2424-2433.
[7] Yan XJ, Xu J, Gu ZH, et al.Exome sequencing identifies somatic mutations of DNA methyltransferase gene DNMT3A in acute monocytic leukemia[J]. Nat Genet,2011, 43(4):309-315.
[8] Döhner H, Weisdorf DJ, Bloomfield CD.Acute myeloid Leukemia[J]. N Engl J Med,2015,373(12):1136-1152.
[9] Fasan A, Haferlach C, Alpermann T, et al.The role of different genetic subtypes of CEBPA mutated AML[J]. Leukemia,2014,28(4):794-803.
[10] Pabst T, Mueller BU, Zhang P, et al.Dominant-negative mutations of CEBPA, encoding CCAAT/enhancer binding protein-alpha (C/EBPalpha), in acute myeloid leukemia[J]. Nat Genet,2001,27(3):263-270.
[11] Timchenko NA, Wilde M, Nakanishi M, et al.CCAAT/enhancer-binding protein alpha (C/EBP alpha) inhibits cell proliferation through the p21 (WAF-1/CIP-1/SDI-1) protein[J]. Genes Dev,1996,10(7):804-815.
[12] Pabst T, Eyholzer M, Fos J, et al.Heterogeneity within AML with CEBPA mutations; only CEBPA double mutations, but not single CEBPA mutations are associated with favourable prognosis[J]. Br J Cancer,2009,100(8):1343-1346.
[13] Arber DA, Orazi A, Hasserjian R, et al.The 2016 revision to the World Health Organization classification of myeloid neoplasms and acute leukemia[J]. Blood,2016, 127(20):2391-2405.
[14] NCCN Cliinical Practice Guidelines in Oncology-----Acute Myeloid Leukemia[R/OL]. https://www.nccn.org/National Comprehensive Cancer Network,NCCN Cliinical Practice Guidelines in Oncology-----Acute Myeloid Leukemia[R/OL]. https://www.nccn.org/National Comprehensive Cancer Network,Inc,2017:25.
[15] Döhner H, Estey E, Grimwade D, et al.Diagnosis and management of AML in adults: 2017 ELN recommendations from an international expert panel[J]. Blood, 2017 ,129(4):424-447.
[16] Ghanem H, Tank N, Tabbara IA.Prognostic implications of genetic aberrations in acute myelogenous leukemia with normal cytogenetics[J]. Am J Hematol,2012,87(1):69-77.
[17] Shin SY, Lee ST, Kim HJ, et al.Mutation profiling of 19 candidate genes in acute myeloid leukemia suggests significance of DNMT3A mutations[J]. Oncotarget,2016,7(34):54825-54837.
[18] Wakita S, Yamaguchi H, Ueki T, et al.Complex molecular genetic abnormalities involving three or more genetic mutations are important prognostic factors for acute myeloid leukemia[J]. Leukemia,2016,30(3):545-554.
[19] Peterlin P, Renneville A, Ben Abdelali R, et al.Impact of additional genetic alterations on the outcome of patients with NPM1-mutated cytogenetically normal acute myeloid leukemia[J]. Haematologica,2015,100(5):e196-e199.
[20] Shen Y, Zhu YM, Fan X, et al.Gene mutation patterns and their prognostic impact in a cohort of 1185 patients with acute myeloid leukemia[J]. Blood,2011,118(20):5593-5603.
[21] Pastore F, Kling D, Hoster E, et al.Long-term follow-up of cytogenetically normal CEBPA-mutated AML[J]. J Hematol Oncol,2014,7:55.
[22] Daniel A, Arber AO, Robert H, et al.The 2016 revision to the World Health Organization classification of myeloid neoplasms and acute leukemia[M]. Lyon: IARC,2016.
[23] Wouters BJ, Löwenberg B, Erpelinck-Verschueren CA, et al.Double CEBPA mutations, but not single CEBPA mutations, define a subgroup of acute myeloid leukemia with a distinctive gene expression profile that is uniquely associated with a favorable outcome[J]. Blood,2009,113(13):3088-3091.
[24] Borthakur G, Kantarjian H, Patel KP, et al.Impact of numerical variation in FMS-like tyrosine kinase receptor 3 internal tandem duplications on clinical outcome in normal karyotype acute myelogenous leukemia[J]. Cancer,2012,118(23):5819-5822.
[25] Renneville A, Boissel N, Nibourel O, et al.Prognostic significance of DNA methyltransferase 3A mutations in cytogenetically normal acute myeloid leukemia: a study by the Acute Leukemia French Association[J]. Leukemia,2012,26(6):1247-1254.
[26] Wang JH, Chen WL, Li JM, et al.Prognostic significance of 2-hydroxyglutarate levels in acute myeloid leukemia in China[J]. Proc Natl Acad Sci U S A,2013,110(42):17017-17022.
[27] Weissmann S, Alpermann T, Grossmann V, et al.Landscape of TET2 mutations in acute myeloid leukemia[J]. Leukemia,2012,26(5):934-942.