DataSheet_1_Up-Regulation of TRIM32 Associated With the Poor Prognosis of Acute Myeloid Leukemia by Integrated Bioinformatics Analysis With External Validation.docx

BackgroundAcute myeloid leukemia (AML) is a malignant and molecularly heterogeneous disease. It is essential to clarify the molecular mechanisms of AML and develop targeted treatment strategies to improve patient prognosis.
MethodsAML mRNA expression data and survival status were extracted from TCGA and GEO databases (GSE37642, GSE76009, GSE16432, GSE12417, GSE71014). Weighted gene co-expression network analysis (WGCNA) and differential gene expression analysis were performed. Functional enrichment analysis and protein-protein interaction (PPI) network were used to screen out hub genes. In addition, we validated the expression levels of hub genes as well as the prognostic value and externally validated TRIM32 with clinical data from our center. AML cell lines transfected with TRIM32 shRNA were also established to detect the proliferation in vitro.
ResultsA total of 2192 AML patients from TCGA and GEO datasets were included in this study and 20 differentially co-expressed genes were screened by WGCNA and differential gene expression analysis methods. These genes were mainly enriched in phospholipid metabolic processes (biological processes, BP), secretory granule membranes (cellular components, CC), and protein serine/threonine kinase activity (molecular functions, MF). In addition, the protein-protein interaction (PPI) network contains 15 nodes and 15 edges and 10 hub genes (TLE1, GLI2, HDAC9, MICALL2, DOCK1, PDPN, RAB27B, SIX3, TRIM32 and TBX1) were identified. The expression of 10 central genes, except TLE1, was associated with survival status in AML patients (p<0.05). High expression of TRIM32 was tightly associated with poor relapse-free survival (RFS) and overall survival (OS) in AML patients, which was verified in the bone marrow samples from our center. In vitro, knockdown of TRIM32 can inhibit the proliferation of AML cell lines.
ConclusionTRIM32 was associated with the progression and prognosis of AML patients and could be a potential therapeutic target and biomarker for AML in the future.

研究背景：急性髓系白血病（Acute myeloid leukemia, AML）是一种恶性且具有分子异质性的疾病。阐明AML的分子机制并开发靶向治疗策略以改善患者预后，具有重要的临床意义。
研究方法：从癌症基因组图谱（The Cancer Genome Atlas, TCGA）及基因表达综合数据库（Gene Expression Omnibus, GEO，包含GSE37642、GSE76009、GSE16432、GSE12417、GSE71014数据集）中提取AML的mRNA表达数据与生存状态信息。开展加权基因共表达网络分析（Weighted gene co-expression network analysis, WGCNA）与差异基因表达分析，通过功能富集分析及蛋白质相互作用（protein-protein interaction, PPI）网络筛选枢纽基因（hub genes）。此外，本研究验证了枢纽基因的表达水平与预后价值，并利用本中心的临床数据对TRIM32进行外部验证；同时构建转染TRIM32短发夹RNA（short hairpin RNA, shRNA）的AML细胞系，开展体外增殖检测。
研究结果：本研究共纳入TCGA与GEO数据集的2192例AML患者，通过WGCNA与差异基因表达分析筛选得到20个差异共表达基因。上述基因主要富集于磷脂代谢过程（生物过程，BP）、分泌颗粒膜（细胞组分，CC）以及蛋白质丝氨酸/苏氨酸激酶活性（分子功能，MF）。蛋白质相互作用（PPI）网络包含15个节点与15条边，最终鉴定出10个枢纽基因（hub genes）：TLE1、GLI2、HDAC9、MICALL2、DOCK1、PDPN、RAB27B、SIX3、TRIM32及TBX1。除TLE1外，其余9个枢纽基因的表达水平均与AML患者的生存状态显著相关（p<0.05）。TRIM32高表达与AML患者较差的无复发生存期（relapse-free survival, RFS）及总生存期（overall survival, OS）密切相关，该结论在本中心的骨髓样本中得到验证。体外实验证实，敲低TRIM32可抑制AML细胞系的增殖。
研究结论：TRIM32与AML患者的疾病进展及预后密切相关，有望成为未来AML治疗的潜在靶点与生物标志物。