骨髓增生异常综合征与急性髓系白血病之间免疫浸润相关基因的筛选及验证. (Chinese)

Screening and verification of genes related to immune infiltration between myelodysplastic syndrome and acute myeloid leukemia. (English)

BACKGROUND: Myelodysplastic syndrome has worse hazards of acute myeloid leukemia transformation, and some studies have revealed that immune infiltration plays a vital part in the two. Nevertheless, more studies are required to confirm the relationship between immune infiltration and related differentially expressed gene regulation. OBJECTIVE: To screen the differentially expressed genes with prognostic significance between myelodysplastic syndrome and acute myeloid leukemia by bioinformatics analysis and explore the possible roles and mechanisms among these differentially expressed genes and immune infiltration mechanisms in the occurrence and progression of diseases. METHODS: The differentially expressed genes were screened for bioinformatics analysis using the GEO datasets, and analyzed by DO, GO, KEGG and GSEA. The TCGA prognostic database was used to plot the K-M curves of differentially expressed genes and receiver operating characteristic curve analysis was applied to evaluate the clinical diagnostic performance. Finally, CIBERSORT analysis was used to intuitively demonstrate the correlation between critical prognostic genes and the distribution of immuno-infiltrated cells. RT-qPCR was employed to detect peripheral blood samples from healthy controls, myelodysplastic syndrome and acute myeloid leukemia patients so as to verify the crucial genes preliminarily. RESULTS AND CONCLUSION: (1) A total of 150 differentially expressed genes were obtained between myelodysplastic syndrome and acute myeloid leukemia, among which 16 genes were up-regulated and 134 were down-regulated. (2) The results of DO, GO, KEGG and GSEA analysis suggested that differentially expressed genes might promote the development of myelodysplastic syndrome to acute myeloid leukemia by regulating the immune response. CIBERSORT revealed the differences in immune infiltration between myelodysplastic syndrome and acute myeloid leukemia. The distribution of CD4⁺ T cells, monocytes, neutrophils and M1 macrophages decreased in acute myeloid leukemia patients. In contrast, the distribution of inflammatory suppressor cells M2 macrophages increased, suggesting that it may be related to the immunosuppression of acute myeloid leukemia. (3) K-M curve and receiver operating characteristic curve analysis of 150 differentially expressed genes screened out four genes relevant to immunity and prognosis with good diagnostic performance: MANSC1, FLT3, BMX and CXCR2. (4) The results of RT-qPCR exhibited that MANSC1, BMX and CXCR2 were low expressed, while FLT3 was highly expressed in acute myeloid leukemia patients. These findings verify that the differential expression of MANSC1, FLT3, BMX and CXCR2 in patients with myelodysplastic syndrome and acute myeloid leukemia is not only significantly correlated with the prognosis of patients but may also affect the occurrence and development of myelodysplastic syndrome and acute myeloid leukemia by regulating the immune infiltration of patients. They can be used as potential biomarkers and therapeutic targets of the transformation from myelodysplastic syndrome to acute myeloid leukemia, providing a new direction for clinical diagnosis and treatment of the transformation of myelodysplastic syndrome. [ABSTRACT FROM AUTHOR]

背景：骨髓增生异常综合征具有较高的急性髓系白血病转化风险, 有研究表明免疫浸润在二者之间发挥重要作用, 但免疫细胞浸润与相关 差异表达基因的调控关系仍需更多研究证实。 目的：通过生物信息学分析方法, 寻找与骨髓增生异常综合征和急性髓系白血病具有预后意义的差异表达基因, 探讨差异基因与免疫浸润 细胞调控在疾病发生发展中的可能作用及机制。 方法：利用GEO数据集筛选差异表达基因用于生物信息学分析, 对差异表达基因进行DO、GO、KEGG和GSEA分析。利用TCGA预后数据库绘 制差异表达基因的K-M曲线, 并对预后相关的差异基因进行受试者工作特征曲线分析判断基因的临床诊断性能。最后, 采用CIBERSORT分 析预后相关关键基因与免疫浸润细胞分布之间的相关性, 利用RT-qPCR检测健康对照、骨髓增生异常综合征及急性髓系白血病患者外周血 样本对筛选出的关键基因进行初步验证。 结果与结论：①在骨髓增生异常综合征和急性髓系白血病之间共获得150个差异表达基因, 其中16个基因表达上调, 134个基因表达下调； ②DO、GO、KEGG及GSEA分析结果提示差异表达基因可能通过调节免疫应答促进骨髓增生异常综合征向急性髓系白血病发展, CIBERSORT 分析显示骨髓增生异常综合征与急性髓系白血病之间存在免疫浸润差异, 其中CD4⁺ T细胞、单核细胞、中性粒细胞、M1巨噬细胞在急性 髓系白血病患者中分布降低, 而炎症抑制细胞M2巨噬细胞在急性髓系白血病患者中分布增加, 可能与急性髓系白血病免疫抑制有关；③ 利用K-M曲线及受试者工作特征曲线从150个差异基因中筛选出4个与免疫和预后相关且具有较好诊断性能的基因：MANSC1、FLT3、BMX及 CXCR2；④RT-qPCR检测结果显示MANSC1、BMX及CXCR2在急性髓系白血病患者中低表达, 而FLT3在急性髓系白血病患者中高表达。结果表 明, MANSC1、FLT3、BMX及CXCR2在骨髓增生异常综合征与急性髓系白血病患者间差异表达, 不仅与患者预后显著相关, 还可能通过调节 患者的免疫浸润影响骨髓增生异常综合征及急性髓系白血病的发生和进展, 它们可作为骨髓增生异常综合征向急性髓系白血病转化的潜在 生物标志物和治疗靶点, 为骨髓增生异常综合征“转白”的临床诊疗研究提供新的方向。 [ABSTRACT FROM AUTHOR]

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