Liu Chang, Jiang Hao, Zhu Andu, Xu Chen, Wang Zhenfan, Mao Guocai, Jiang Minjun, Chen Jianchun, Ma Zheng, Qi Jiaqian, Cao Zhijun
Department of Urology, Suzhou Ninth People's Hospital, Soochow University, Suzhou, China.
Department of Urology, The First Affiliated Hospital of Soochow University, Suzhou, China.
Front Genet. 2025 Jun 13;16:1588941. doi: 10.3389/fgene.2025.1588941. eCollection 2025.
End-stage renal disease (ESRD) is increasing worldwide, and although kidney transplantation improves survival, long-term graft loss-driven mainly by immune-mediated rejection-remains common. We aimed to delineate immune mechanisms that distinguish recipients with stable versus impaired graft function.
Peripheral blood mononuclear cells from kidney-transplant recipients with normal (n = 10) or impaired (n = 10) renal function were profiled by single-cell RNA sequencing. Fourteen immune populations were identified; CD4 T-cell "stemness" was quantified using mRNAsi and EREG_mRNAsi indices, lineage trajectories were reconstructed with Monocle, and ligand-receptor communication was inferred with iTalk. Findings were validated in an independent bulk RNA-seq cohort (n = 192) using differential expression and weighted gene co-expression network analysis (WGCNA).
Recipients with graft dysfunction exhibited (i) expansion of Th17 cells and contraction of Treg cells, (ii) significant loss of CD4 T-cell stem-like features (lower mRNAsi/EREG_mRNAsi, p < 0.001), and (iii) pseudotime trajectories skewed toward Th17 differentiation. iTalk revealed enhanced S100A8/A9-TLR4 signalling from myeloid cells to neutrophils, consistent with reduced circulating neutrophils and presumptive intragraft accumulation. Bulk validation confirmed the stemness deficit and identified eight hub genes (API5, CAPRIN1, CCT2, DLG1, NMD3, RDX, SENP7, S100A4) that correlated with both low stemness and poor clinical outcome. Pathway enrichment implicated cell-morphogenesis, tight-junction, and metabolic-homeostasis pathways in graft injury.
Integrative single-cell and bulk analyses link diminished CD4 T-cell stemness, Th17-dominant polarization, and S100A4-mediated neutrophil recruitment to graft dysfunction. These signatures nominate stemness indices, Th17/Treg balance, and the S100-TLR4 axis as candidate biomarkers and therapeutic targets to preserve allograft integrity and prolong transplant survival.
终末期肾病（ESRD）在全球范围内呈上升趋势，尽管肾移植可提高生存率，但主要由免疫介导的排斥反应导致的长期移植物丢失仍然很常见。我们旨在阐明区分移植肾功能稳定与受损受者的免疫机制。
通过单细胞RNA测序对肾功能正常（n = 10）或受损（n = 10）的肾移植受者的外周血单个核细胞进行分析。鉴定出14种免疫细胞群；使用mRNAsi和EREG_mRNAsi指数对CD4 T细胞的“干性”进行量化，用Monocle重建细胞谱系轨迹，并用iTalk推断配体-受体通讯。在一个独立的批量RNA测序队列（n = 192）中，使用差异表达和加权基因共表达网络分析（WGCNA）对研究结果进行验证。
移植功能障碍的受者表现为：（i）Th17细胞扩增和调节性T细胞收缩；（ii）CD4 T细胞干性样特征显著丧失（mRNAsi/EREG_mRNAsi降低，p < 0.001）；（iii）拟时间轨迹偏向Th17分化。iTalk显示从髓样细胞到中性粒细胞的S100A8/A9-TLR4信号增强，这与循环中性粒细胞减少和假定的移植物内积聚一致。批量验证证实了干性缺陷，并鉴定出8个中心基因（API5、CAPRIN1、CCT2、DLG1、NMD3、RDX、SENP7、S100A4），这些基因与低干性和不良临床结局均相关。通路富集表明细胞形态发生、紧密连接和代谢稳态通路与移植物损伤有关。
综合单细胞和批量分析表明，CD4 T细胞干性降低、Th17主导的极化以及S100A4介导的中性粒细胞募集与移植物功能障碍有关。这些特征将干性指数、Th17/Treg平衡以及S100-TLR4轴确定为候选生物标志物和治疗靶点，以维持同种异体移植物的完整性并延长移植存活时间。
