CEBPA-double-mutated acute myeloid leukemia displays a unique phenotypic profile: a reliable screening method and insight into biological features.
CEBPA-double-mutated acute myeloid leukemia displays a unique phenotypic profile: a reliable screening method and insight into biological features.
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DOI：
10.3324/haematol.2016.151910
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发表时间：
2017-03
期刊：
影响因子：
10.1
通讯作者：
Bosi A
中科院分区：
文献类型：
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作者：
Mannelli F;Ponziani V;Bencini S;Bonetti MI;Benelli M;Cutini I;Gianfaldoni G;Scappini B;Pancani F;Piccini M;Rondelli T;Caporale R;Gelli AM;Peruzzi B;Chiarini M;Borlenghi E;Spinelli O;Giupponi D;Zanghì P;Bassan R;Rambaldi A;Rossi G;Bosi A
5-10% 的急性髓系白血病病例中会出现 CCAAT/增强子结合蛋白 α (CEBPA) 突变。 CEBPA 双突变病例通常携带双等位基因 N 端和 C 端突变，并与良好的临床结果相关。由于突变的多样性、基因的内在特征和技术问题，CEBPA 突变体的鉴定具有挑战性。已经提出了几种专门针对大规模临床使用的筛选方法（片段长度分析、基因表达阵列）；尽管有效，但它们受到具体问题的限制。我们研究了 251 例急性髓系白血病诊断时原始细胞和成熟骨髓细胞区室的表型特征。在该队列中，16 名患者 (6.4%) 具有两种 CEBPA 突变，而 10 名患者 (4.0%) 具有单一突变。首先，我们强调 CEBPA 双突变子集在所有细胞区室中显示出反复出现的表型异常。通过细胞分选后的突变分析，我们证明了这种常见的表型特征取决于 CEBPA 双突变的多谱系参与。通过对表型数据的多维研究，我们开发了一个包括十个核心且广泛可用的参数的分类器。原始细胞（CD34、CD117、CD7、CD15、CD65）、中性粒细胞（SSC、CD64）、单核细胞（CD14、CD64）和红细胞（CD117）区室上选定的标记能够对 CEBPA 双突变病例进行聚类。在来自三个独立中心的 259 例 AML 病例验证集中，我们的分类器表现出优异的性能，具有 100% 的特异性和 100% 的敏感性。因此，我们基于对广泛可用的表型参数的多维分析，建立了一种可靠的筛选方法。该方法提供了早期结果，适合大规模检测CEBPA双突变状态，使基因测序能够集中于选定的病例。
Mutations in CCAAT/enhancer binding protein α (CEBPA) occur in 5–10% of cases of acute myeloid leukemia. CEBPA-double-mutated cases usually bear biallelic N- and C-terminal mutations and are associated with a favorable clinical outcome. Identification of CEBPA mutants is challenging because of the variety of mutations, intrinsic characteristics of the gene and technical issues. Several screening methods (fragment-length analysis, gene expression array) have been proposed especially for large-scale clinical use; although efficient, they are limited by specific concerns. We investigated the phenotypic profile of blast and maturing bone marrow cell compartments at diagnosis in 251 cases of acute myeloid leukemia. In this cohort, 16 (6.4%) patients had two CEBPA mutations, whereas ten (4.0%) had a single mutation. First, we highlighted that the CEBPA-double-mutated subset displays recurrent phenotypic abnormalities in all cell compartments. By mutational analysis after cell sorting, we demonstrated that this common phenotypic signature depends on CEBPA-double-mutated multi-lineage involvement. From a multidimensional study of phenotypic data, we developed a classifier including ten core and widely available parameters. The selected markers on blasts (CD34, CD117, CD7, CD15, CD65), neutrophil (SSC, CD64), monocytic (CD14, CD64) and erythroid (CD117) compartments were able to cluster CEBPA-double-mutated cases. In a validation set of 259 AML cases from three independent centers, our classifier showed excellent performance with 100% specificity and 100% sensitivity. We have, therefore, established a reliable screening method, based upon multidimensional analysis of widely available phenotypic parameters. This method provides early results and is suitable for large-scale detection of CEBPA-double-mutated status, allowing gene sequencing to be focused in selected cases.