单细胞 RNA 和 ATAC 测序揭示了血液透析相关的循环免疫细胞亚群免疫失调。
Single-Cell RNA and ATAC Sequencing Reveal Hemodialysis-Related Immune Dysregulation of Circulating Immune Cell Subpopulations.
机构信息
The Second Clinical Medical College of Jinan University, Shenzhen People's Hospital, Shenzhen, China.
Institute of Nephrology and Blood Purification, The First Affiliated Hospital of Jinan University, Jinan University, Guangzhou, China.
出版信息
Front Immunol. 2022 May 26;13:878226. doi: 10.3389/fimmu.2022.878226. eCollection 2022.
BACKGROUND
An increased risk of infection, malignancy, and cardiovascular diseases in maintenance hemodialysis patients is associated with hemodialysis-related immunity disturbances. Although defects in T-lymphocyte-dependent immune responses and preactivation of antigen-presenting cells have been documented in hemodialysis patients, the effects of long-term hemodialysis on the transcriptional program and chromosomal accessibility of circulating immune cell subpopulations remain poorly defined.
METHODS
We integrated single-cell RNA sequencing (scRNA-seq) and single-cell assay for transposase-accessible chromatin sequencing (scATAC-seq) to characterize the transcriptome profiles of peripheral mononuclear cells (PBMCs) from healthy controls and maintenance hemodialysis patients. Validation of differentially expressed genes in CD4+ T cells and monocytes were performed by magnetic bead separation and quantitative real-time PCR.
RESULTS
We identified 16 and 15 PBMC subgroups in scRNA-seq and scATAC-seq datasets, respectively. Hemodialysis significantly suppressed the expression levels of T cell receptor (TCR) genes in CD4+ T cell subsets (e.g., TRAV4, CD45, CD3G, CD3D, CD3E) and major histocompatibility complex II (MHC-II) pathway-related genes in monocytes (HLA-DRB1, HLA-DQA2, HLA-DQA1, HLA-DPB1). Downstream pathways of TCR signaling, including PI3K-Akt-mTOR, MAPK, TNF, and NF-κB pathways, were also inhibited in CD4+ T cell subpopulations during the hemodialysis procedure. Hemodialysis altered cellular communication patterns between PBMC subgroups, particularly TGF-TGFBR, HVEM-BTLA, and IL16-CD4 signalings between CD4+ T cells and monocytes. Additionally, we found that hemodialysis inhibited the expression of AP-1 family transcription factors (JUN, JUND, FOS, FOSB) by interfering with the chromatin accessibility profile.
CONCLUSIONS
Our study provides a valuable framework for future investigations of hemodialysis-related immune dysregulation and identifies potential therapeutic targets for reconstituting the circulating immune system in maintenance hemodialysis patients.
背景
维持性血液透析患者感染、恶性肿瘤和心血管疾病的风险增加与血液透析相关的免疫紊乱有关。尽管已经在血液透析患者中记录了 T 淋巴细胞依赖性免疫应答缺陷和抗原呈递细胞的预激活，但长期血液透析对循环免疫细胞亚群的转录程序和染色质可及性的影响仍知之甚少。
方法
我们整合了单细胞 RNA 测序（scRNA-seq）和单细胞转座酶可及染色质测序（scATAC-seq），以描述健康对照者和维持性血液透析患者外周血单个核细胞（PBMC）的转录组谱。通过磁珠分离和实时定量 PCR 验证 CD4+T 细胞和单核细胞中差异表达基因。
结果
我们在 scRNA-seq 和 scATAC-seq 数据集中分别鉴定出 16 个和 15 个 PBMC 亚群。血液透析显著抑制了 CD4+T 细胞亚群中 T 细胞受体（TCR）基因（例如 TRAV4、CD45、CD3G、CD3D、CD3E）和单核细胞中主要组织相容性复合体 II（MHC-II）途径相关基因（HLA-DRB1、HLA-DQA2、HLA-DQA1、HLA-DPB1）的表达水平。TCR 信号转导的下游途径，包括 PI3K-Akt-mTOR、MAPK、TNF 和 NF-κB 途径，也在血液透析过程中被抑制。血液透析改变了 PBMC 亚群之间的细胞通讯模式，特别是 CD4+T 细胞和单核细胞之间的 TGF-TGFBR、HVEM-BTLA 和 IL16-CD4 信号通路。此外，我们发现血液透析通过干扰染色质可及性谱抑制了 AP-1 家族转录因子（JUN、JUND、FOS、FOSB）的表达。
结论
本研究为进一步研究血液透析相关免疫失调提供了有价值的框架，并确定了维持性血液透析患者重建循环免疫系统的潜在治疗靶点。
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