Table_1_Pre-transplant Transcriptional Signature in Peripheral Blood Mononuclear Cells of Acute Renal Allograft Rejection.XLSX

Acute rejection (AR) is closely associated with renal allograft dysfunction. Here, we utilised RNA sequencing (RNA-Seq) and bioinformatic methods to characterise the peripheral blood mononuclear cells (PBMCs) of patients with acute renal allograft rejection. Pretransplant blood samples were collected from 32 kidney allograft donors and 42 corresponding recipients with biopsies classified as T cell-mediated rejection (TCMR, n = 18), antibody-mediated rejection (ABMR, n = 5), and normal/non-specific changes (non-AR, n = 19). The patients with TCMR and ABMR were assigned to the AR group, and the patients with normal/non-specific changes (n = 19) were assigned to the non-AR group. We analysed RNA-Seq data for identifying differentially expressed genes (DEGs), and then gene ontology (GO) analysis, Reactome, and ingenuity pathway analysis (IPA), protein—protein interaction (PPI) network, and cell-type enrichment analysis were utilised for bioinformatics analysis. We identified DEGs in the PBMCs of the non-AR group when compared with the AR, ABMR, and TCMR groups. Pathway and GO analysis showed significant inflammatory responses, complement activation, interleukin-10 (IL-10) signalling pathways, classical antibody-mediated complement activation pathways, etc., which were significantly enriched in the DEGs. PPI analysis showed that IL-10, VEGFA, CXCL8, MMP9, and several histone-related genes were the hub genes with the highest degree scores. Moreover, IPA analysis showed that several proinflammatory pathways were upregulated, whereas antiinflammatory pathways were downregulated. The combination of NFSF14+TANK+ANKRD 33 B +HSPA1B was able to discriminate between AR and non-AR with an AUC of 92.3% (95% CI 82.8–100). Characterisation of PBMCs by RNA-Seq and bioinformatics analysis demonstrated gene signatures and biological pathways associated with AR. Our study may provide the foundation for the discovery of biomarkers and an in-depth understanding of acute renal allograft rejection.

急性排斥反应（AR）与肾脏同种异体移植功能障碍密切相关。本研究中，我们利用RNA测序（RNA-Seq）和生物信息学方法对急性肾脏同种异体排斥反应患者的周围血单个核细胞（PBMCs）进行了特征分析。收集了32名肾脏同种异体移植供体的移植前血液样本和42名相应受体的活检样本，其中T细胞介导的排斥反应（TCMR，n=18）、抗体介导的排斥反应（ABMR，n=5）和正常/非特异性变化（非AR，n=19）被分类。TCMR和ABMR的患者被分配到AR组，而正常/非特异性变化的患者（n=19）被分配到非AR组。我们分析了RNA-Seq数据以识别差异表达基因（DEGs），随后利用基因本体（GO）分析、Reactome、创新路径分析（IPA）、蛋白质-蛋白质相互作用（PPI）网络和细胞类型富集分析进行了生物信息学分析。与非AR组相比，我们在AR、ABMR和TCMR组的PBMCs中识别出DEGs。通路和GO分析显示出显著的炎症反应、补体激活、白细胞介素-10（IL-10）信号通路、经典抗体介导的补体激活通路等，这些通路在DEGs中显著富集。PPI分析显示IL-10、VEGFA、CXCL8、MMP9和若干组蛋白相关基因是具有最高度分数的中心基因。此外，IPA分析显示，多个促炎通路上调，而抗炎通路下调。NFSF14+TANK+ANKRD 33 B +HSPA1B的组合能够以92.3%的AUC（95% CI 82.8–100）区分AR与非AR。通过RNA-Seq和生物信息学分析对PBMCs进行特征分析，揭示了与AR相关的基因特征和生物学通路。本研究可能为生物标志物的发现以及对急性肾脏同种异体排斥反应的深入理解奠定基础。