研究尚未系统比较急性髓细胞白血病患者配对的单核细胞与具有活力的CD34表达的白血病飞蚊症的预后转录本的性能确认能力。我们猜测检查同质性的白血病飞蚊症将产生不同的生物信息，并可能提高表达生物标志物的预后性能，为了评估细胞异质性对急性髓细胞白血病中的表达生物标志物的影响，我们系统地检查了SWOG诊断标本中的配对单核细胞和具有活力的CD34表达的白血病飞蚊症。患者经过富集后，根据提取的RNA的可用性，分配到发现和验证群。RNA测序数据的分析研究了如何富集对与预分析变量、患者特征和临床结局相关的差异表达基因产生影响。飞蚊症富集产生了显著不同的表达谱和与临床特征（例如细胞遗传学）相关的生物通路。虽然许多差异表达基因与临床结局相关，但经过年龄和ELN危险调整后，在单核细胞和飞蚊症中，大多数基因在预后上失去了它们的预测意义，只有11个基因在两种细胞群体中的总生存中仍然显著（CEP70，COMMD7，DNMT3B，ECE1，LNX2，NEGR1，PIK3C2B，SEMA4D，SMAD2，TAF8，ZNF444）。为了检查富集对生物标记验证的影响，这11个候选生物标志物在验证群中通过定量RT / PCR进行了研究。经过年龄和ELN危险调整后，4个基因（CEP70，DNMT3B，ECE1和PIK3CB）的表达在飞蚊症中仍与总生存显着相关，而在单核细胞中没有一个基因达到统计显着。本研究为研究同一患者的单核细胞和飞蚊症可能获得/失去的生物信息提供了见解，并显示了飞蚊症中表达生物标志物的改进验证率。 ©2023年 作者（们）。

Studies have not systematically compared the ability to verify performance of prognostic transcripts in paired bulk mononuclear cells versus viable CD34-expressing leukemic blasts from patients with acute myeloid leukemia. We hypothesized that examining the homogenous leukemic blasts will yield different biological information and may improve prognostic performance of expression biomarkers.To assess the impact of cellular heterogeneity on expression biomarkers in acute myeloid leukemia, we systematically examined paired mononuclear cells and viable CD34-expressing leukemic blasts from SWOG diagnostic specimens. After enrichment, patients were assigned into discovery and validation cohorts based on availability of extracted RNA. Analyses of RNA sequencing data examined how enrichment impacted differentially expressed genes associated with pre-analytic variables, patient characteristics, and clinical outcomes.Blast enrichment yielded significantly different expression profiles and biological pathways associated with clinical characteristics (e.g., cytogenetics). Although numerous differentially expressed genes were associated with clinical outcomes, most lost their prognostic significance in the mononuclear cells and blasts after adjusting for age and ELN risk, with only 11 genes remaining significant for overall survival in both cell populations (CEP70, COMMD7, DNMT3B, ECE1, LNX2, NEGR1, PIK3C2B, SEMA4D, SMAD2, TAF8, ZNF444). To examine the impact of enrichment on biomarker verification, these 11 candidate biomarkers were examined by quantitative RT/PCR in the validation cohort. After adjusting for ELN risk and age, expression of 4 genes (CEP70, DNMT3B, ECE1, and PIK3CB) remained significantly associated with overall survival in the blasts, while none met statistical significance in mononuclear cells.This study provides insights into biological information gained/lost by examining viable CD34-expressing leukemic blasts versus mononuclear cells from the same patient and shows an improved verification rate for expression biomarkers in blasts.© 2023. The Author(s).