基于CML伊马替尼耐药相关lncRNA的筛选及风险预测模型研究

Imatinib mesylate, a selective tyrosine kinase inhibitor of Bcr/Abl, is regarded as the primary specific chemotherapeutic agent against chronic myeloid leukemia (CML). However, resistance to imatinib poses a major problem for the treatment of CML nowadays. It is estimated that the mechanisms of imatinib resistance remain unknown in about 30-50% of patients. Coding sequence has been the focus of the mechanism studies so far, while relationships between long noncoding RNA (lncRNA) and imatinib resistance have been rarely reported. In our previous work, lncRNAs were discovered to be involved in imatinib resistance by functional sequence mutations of parental genes. This study is designed to screen and identify lncRNAs involved in imatinib resistance, by bioinformatics and molecular cell biology technology (targeted high throughput sequencing, co-expression network analysis, etc). A mathematical model, based on lncRNAs involved in imatinib resistance and BCR/ABL cDNA, would be further derived to predict imatinib resistance risk for CML. This study may help to elucidate new molecular mechanisms for imatinib resistance and develop mathematical models to predict imatinib resistance risk, thus to provide scientific bases for clinical treatments.

近年来, 慢性髓性白血病（CML）患者对伊马替尼(CML的一线靶向治疗药物)耐药的问题愈加严重，其中约30-50%患者的伊马替尼耐药机制不明。目前，CML耐药机制的研究主要集中于基因组蛋白编码区，而长链非编码RNA（lncRNA）与CML耐药的相关研究还鲜有报道。本课题组前期通过对20例BCR/ABL激酶区突变检测结果为阴性的CML耐药患者样本进行测序分析，发现lncRNA可以通过亲本基因的功能基序突变参与调控伊马替尼耐药。本项目拟通过生物信息学技术以及高通量靶向测序、lncRNA-mRNA共表达等分子生物学技术，筛选并鉴定伊马替尼耐药相关的lncRNA，建立基于BCR/ABL cDNA联合lncRNA序列变异预测伊马替尼耐药的模型，从而实现对CML患者的伊马替尼耐药风险的早期评估，并有望发现伊马替尼耐药lncRNA分子标志物，为完善CML的临床治疗策略提供理论依据。

慢性粒细胞白血病（CML）是一种克隆性造血干细胞疾病，约20-30％的CML患者在接受甲磺酸伊马替尼（IM）治疗时会产生耐药性。这项研究旨在筛选IM耐药相关候选生物标志物，可基于此进一步针对CML患者提供有效治疗策略。本研究从Gene Expression Omnibus（GEO）数据库获取IM耐药或IM敏感的CML患者的基因表达谱，数据预处理后，确定两组之间差异表达的mRNA（DEmRNA），lncRNA（DElncRNA）和microRNA（DEmiRNA），然后进行Gene Ontology （GO）分析和Kyoto Encyclopedia of Genes and Genomes (KEGG) 分析。然后，我们进一步构建了ceRNA网络和蛋白质-蛋白质相互作用网络（PPI），累计发现了49个DElncRNA（12个上调和37个下调DElncRNA），59个DEmiRNA（34个上调和25个下调DEmiRNA）和837个DEmRNA（510个上调和327个下调DEmRNA），建立了IM 耐药相关lncRNA-miRNA-mRNA共调控网络，提示PI3K-Akt信号通路和Ras信号通路参与IM耐药。此外，鉴定出6个hub 基因，包括VEGFA，NRAS，CREB1，STAT1，NPM1和PIK3R1。MIAT-VEGFA可能在CML的预后中起关键作用。总而言之，当前研究为IM耐药共调控网络提供了一个全面的视角，并且确定了CML的早期风险评估和预后评估的潜在分子生物标志物。