单倍体造血干细胞移植后肠道急性移植物抗宿主病的发病机制

Pathogenesis of intestinal acute graft-versus-host disease after haploid hematopoietic stem cell transplantation

背景:肠道急性移植物抗宿主病是异基因造血干细胞移植后最常见的危及生命的并发症之一,由供体免疫活性T细胞攻击健康受体组织导致,目前治疗方案效果不佳。长链非编码RNA参与免疫反应等多种生物学过程,但在肠道急性移植物抗宿主病中的作用尚不清楚,有待进一步研究。目的:构建肠道急性移植物抗宿主病差异长链非编码RNA和mRNA表达谱,预测其可能的作用及机制。方法:采集4例单倍体造血干细胞移植后肠道急性移植物抗宿主病患者和4名健康志愿者的外周血单个核细胞进行高通量测序,并对差异mRNA进行GO和KEGG富集分析。根据基因表达信号值的动态变化,构建差异长链非编码RNA-mRNA共表达网络,基于长链非编码RNA对下游靶基因不同的调节机制,分别预测其通过顺式或反式调控及经由竞争性内源RNA机制调控的靶基因,使用Cytoscape构建基因互作网络,通过对靶基因的功能分析预测差异长链非编码RNA可能的作用及相关机制。结果与结论:肠道急性移植物抗宿主病患者的外周血单个核细胞中有1311个长链非编码RNA和3283个mRNA差异表达,GO和KEGG结果提示这些差异mRNA主要富集在白细胞介素18、白细胞介素1介导的信号通路、转录因子AP-1复合物、巨核细胞分化和肿瘤坏死因子信号通路,这与肠道炎性反应及与骨髓造血能力关系密切。差异长链非编码RNA顺式及反式调控的靶基因包括一些炎症相关分子,如ZEB2、CDK6、CEACAM1、CXCL1等,经由竞争性内源RNA机制调控的通路主要有PI3K/Akt通路和Notch通路,这与T细胞的活化以及肠道干细胞的稳态相关。结果表明,差异长链非编码RNA可能通过调控炎症相关因子的表达,影响T细胞介导的免疫反应及肠道干细胞的稳态,在肠道急性移植物抗宿主病的发病及进展中发挥重要作用,可能成为诊断及治疗的新靶点。

BACKGROUND:Acute graft-versus-host disease is one of the most common life-threatening complications of allogeneic hematopoietic stem cell,caused by the attack of donor immunoreactive T cells on healthy recipient tissue.The current treatment plan is not effective.Long non-coding RNAs(lncRNAs)are involved in many biological processes such as immune response,but their roles in acute graft-versus-host disease remain unclear and need to be further studied.OBJECTIVE:To construct profiles of differentially expressed lncRNAs and differentially expressed mRNAs of intestinal acute graft-versus-host disease and predict their possible roles and mechanisms.METHODS:Peripheral blood mononuclear cells from four patients with intestinal acute graft-versus-host disease after haploidentical hematopoietic stem cell transplantation and four healthy volunteers were collected for high-throughput sequencing.GO and KEGG enrichment analyses were performed on the differentially expressed mRNAs.According to the dynamic change of gene expression signal value,the differential lncRNA-mRNA co-expression network was constructed.Based on the different regulatory mechanisms of lncRNA on downstream target genes,the target genes regulated by cis or trans regulation and competitive endogenous RNA mechanisms were predicted,respectively.Cytoscape was used to construct the gene interaction networks.The possible effects and related mechanisms of differentially expressed lncRNAs were predicted through functional analysis of their target genes.RESULTS AND CONCLUSION:There are 1311 lncRNAs and 3283 mRNAs differentially expressed in the peripheral blood mononuclear cells of patients with intestinal acute graft-versus-host disease.GO and KEGG analyses indicated that these differentially expressed mRNAs were involved in interleukin-18 or interleukin-1 mediated signaling pathway,AP-1 complex,megakaryocyte differentiation,and tumor necrosis factor signaling pathway,which were closely related to intestinal inflammatory response and hematopoietic ability of bone marrow.Cis-and trans-regulated genes of differentially expressed lncRNAs include some inflammation-related molecules,such as ZEB2,CDK6,CEACAM1,and CXCL1.The pathways regulated by differentially expressed lncRNAs via competitive endogenous RNA mechanism mainly include PI3K/Akt pathway and Notch pathway,which are related to the activation of T cells and the homeostasis of intestinal stem cells.It is concluded that differentially expressed lncRNAs are essential in the pathogenesis and progression of intestinal acute graft-versus-host disease via regulating the expression of inflammation-related molecules,T cell-mediated immune response,and the homeostasis of intestinal stem cells,and may become novel targets for the diagnosis and treatment of intestinal acute graft-versus-host disease.