Gyurina Katalin, Kárai Bettina, Ujfalusi Anikó, Hevessy Zsuzsanna, Barna Gábor, Jáksó Pál, Pálfi-Mészáros Gyöngyi, Póliska Szilárd, Scholtz Beáta, Kappelmayer János, Zahuczky Gábor, Kiss Csongor
Department of Pediatrics, University of Debrecen, Debrecen, Hungary.
Department of Laboratory of Medicine, University of Debrecen, Debrecen, Hungary.
Front Oncol. 2019 Oct 25;9:1063. doi: 10.3389/fonc.2019.01063. eCollection 2019.
Leukemic B-cell precursor (BCP) lymphoblasts were identified as a novel expression site for coagulation factor XIII subunit A (FXIII-A). Flow cytometry (FC) revealed three distinct expression patterns, i.e., FXIII-A negative, FXIII-A dim, and FXIII-A bright subgroups. The FXIII-A negative subgroup was significantly associated with the "B-other" genetic category and had an unfavorable disease outcome. RNA was extracted from bone marrow lymphoblasts of 42 pediatric patients with BCP-acute lymphoblastic leukemia (ALL). FXIII-A expression was determined by multiparameter FC. Genetic diagnosis was based on conventional cytogenetic method and fluorescence hybridization. Affymetrix GeneChip Human Primeview array was used to analyze global expression pattern of 28,869 well-annotated genes. Microarray data were analyzed by Genespring GX14.9.1 software. Gene Ontology analysis was performed using Cytoscape 3.4.0 software with ClueGO application. Selected differentially expressed genes were validated by RT-Q-PCR. We demonstrated, for the first time, the general expression of gene in pediatric BCP-ALL samples. The intensity of expression corresponded to the FXIII-A protein expression subgroups which defined three characteristic and distinct gene expression signatures detected by Affymetrix oligonucleotide microarrays. Relative gene expression intensity of , and followed the pattern of change in the intensity of the expression of the gene. Common enhancer elements of these genes revealed by analysis suggest that common transcription factors may regulate the expression of these genes in a similar fashion. was downregulated in the FXIII-A bright subgroup. Gene expression signature of the FXIII-A negative subgroup showed an overlap with the signature of "B-other" samples. , and were upregulated and was downregulated in the "B-other" subgroup. Validated genes proved biologically and clinically relevant. We described differential expression of genes not shown previously to be associated with pediatric BCP-ALL. Gene expression signature according to FXIII-A protein expression status defined three novel subgroups of pediatric BCP-ALL. Multiparameter FC appears to be an easy-to-use and affordable method to help in selecting FXIII-A negative patients who require a more elaborate and expensive molecular genetic investigation to design precision treatment.
白血病B细胞前体（BCP）淋巴母细胞被确定为凝血因子XIII亚基A（FXIII-A）的一个新表达位点。流式细胞术（FC）揭示了三种不同的表达模式，即FXIII-A阴性、FXIII-A弱阳性和FXIII-A强阳性亚组。FXIII-A阴性亚组与“B-其他”基因类别显著相关，且疾病预后不良。从42例BCP急性淋巴细胞白血病（ALL）儿科患者的骨髓淋巴母细胞中提取RNA。通过多参数FC测定FXIII-A表达。基因诊断基于传统细胞遗传学方法和荧光杂交。使用Affymetrix GeneChip Human Primeview阵列分析28869个注释良好的基因的全局表达模式。微阵列数据通过Genespring GX14.9.1软件进行分析。使用带有ClueGO应用程序的Cytoscape 3.4.0软件进行基因本体分析。通过RT-Q-PCR验证选定的差异表达基因。我们首次证明了该基因在儿科BCP-ALL样本中的普遍表达。其表达强度与FXIII-A蛋白表达亚组相对应，后者定义了通过Affymetrix寡核苷酸微阵列检测到的三种特征性且不同的基因表达特征。、和的相对基因表达强度遵循基因表达强度的变化模式。分析揭示的这些基因的共同增强子元件表明，共同的转录因子可能以类似方式调节这些基因的表达。在FXIII-A强阳性亚组中下调。FXIII-A阴性亚组的基因表达特征与“B-其他”样本的特征有重叠。在“B-其他”亚组中、和上调而下调。经验证的基因在生物学和临床上均具有相关性。我们描述了先前未显示与儿科BCP-ALL相关的基因的差异表达。根据FXIII-A蛋白表达状态的基因表达特征定义了儿科BCP-ALL的三个新亚组。多参数FC似乎是一种易于使用且经济实惠的方法，有助于选择需要更精细且昂贵的分子遗传学研究以设计精准治疗的FXIII-A阴性患者。
