Department of Rheumatology and Immunology, Henan Provincial People's Hospital, People's Hospital of Zhengzhou University, People's Hospital of Henan University, Zhengzhou, China.
Department of Emergency, Henan Provincial People's Hospital, People's Hospital of Zhengzhou University, People's Hospital of Henan University, Zhengzhou, China.
Sci Rep. 2022 May 19;12(1):8398. doi: 10.1038/s41598-022-12594-7.
Rheumatoid arthritis (RA) is an autoimmune disease characterized by persistent synovitis, in which T helper 1 (Th1) can promote the development of a pro-inflammatory microenvironment. Poly(rC)-binding protein 1 (PCBP1) has been identified as a promising biomarker of RA, while its molecular mechanisms in RA development are unknown. As a canonical RNA binding protein, we propose that PCBP1 could play roles in RA by affecting both expression and alternative splicing levels in Th1 cells. Here, microarray datasets (GSE15573 and GSE23561), including 102 peripheral blood mononuclear cell samples from 39 RA patients and 63 controls, were used to evaluate the PCBP1 expression changes in RA patients. High throughput sequencing data (GSE84702) of iron driven pathogenesis in Th1 cells were downloaded and reanalyzed, including two Pcbp1 deficiency samples and two control samples in Th1 cells. In addition, CLIP-seq data of PCBP1 in Jurkat T cells was also analyzed to investigate the regulatory mechanisms of PCBP1. We found PCBP1 were down-regulated in RA specimens compared with control. The result of differentially expressed genes (DEGs) showed that Pcbp1 silencing in Th1 cells affected the expression of genes involved in immune response pathway. Alternative splicing analysis also revealed that PCBP1-regulated alternative splicing genes (RASGs) were enriched in TNF-a/NF-κB signaling pathway, T cell activation, T cell differentiation and T cell differentiation associated immune response pathways, which were highly associated with RA. DEGs and RASGs by Pcbp1 deficiency in mice were validated in PBMCs specimens of RA patients by RT-qPCR. Investigation of the CLIP-seq data revealed PCBP1 preferred to bind to 3'UTR and intron regions. PCBP1-bound genes were also significantly associated with RASGs, identifying 102 overlapped genes of these two gene sets. These genes were significantly enriched in several immune response related pathways, including myeloid cell differentiation and positive regulation of NF-κB transcription factor activity. Two RA-related genes, PML and IRAK1, were screened from the above immune related pathways. These results together support our hypothesis that PCBP1 can regulate the expression of genes involved in immune response pathway, and can bind to and regulate the alternative splicing of immune response related genes in immune T cells, and ultimately participate in the molecular mechanism of RA, providing new research ideas and directions for clinical diagnosis and treatment.
类风湿关节炎(RA)是一种以持续性滑膜炎为特征的自身免疫性疾病,其中辅助性 T 细胞 1(Th1)可以促进促炎微环境的发展。多聚(rC)结合蛋白 1(PCBP1)已被鉴定为 RA 的有前途的生物标志物,但其在 RA 发展中的分子机制尚不清楚。作为一种典型的 RNA 结合蛋白,我们假设 PCBP1 可以通过影响 Th1 细胞的表达和选择性剪接水平来发挥作用。在这里,使用包含 39 名 RA 患者和 63 名对照的 102 个外周血单核细胞样本的微阵列数据集(GSE15573 和 GSE23561)来评估 RA 患者中 PCBP1 的表达变化。下载并重新分析了包括 Th1 细胞中铁驱动发病机制的高通量测序数据(GSE84702),其中包括两个 Pcbp1 缺陷样本和两个对照样本。此外,还分析了 Jurkat T 细胞中 PCBP1 的 CLIP-seq 数据,以研究 PCBP1 的调控机制。我们发现 RA 标本中 PCBP1 的表达低于对照。差异表达基因(DEGs)的结果表明,Th1 细胞中 Pcbp1 的沉默影响了参与免疫反应途径的基因的表达。选择性剪接分析还表明,PCBP1 调节的选择性剪接基因(RASGs)在 TNF-a/NF-κB 信号通路、T 细胞激活、T 细胞分化和 T 细胞分化相关免疫反应途径中富集,这些途径与 RA 高度相关。通过 RT-qPCR 在 RA 患者的 PBMC 标本中验证了 Pcbp1 缺陷小鼠中的 DEGs 和 RASGs。CLIP-seq 数据的研究揭示了 PCBP1 更喜欢结合 3'UTR 和内含子区域。PCBP1 结合的基因也与 RASGs 显著相关,鉴定出这两组基因的 102 个重叠基因。这些基因在几个免疫反应相关途径中显著富集,包括髓样细胞分化和 NF-κB 转录因子活性的正调节。从上述免疫相关途径中筛选出两个 RA 相关基因,PML 和 IRAK1。这些结果共同支持我们的假设,即 PCBP1 可以调节参与免疫反应途径的基因的表达,并可以结合并调节免疫 T 细胞中免疫反应相关基因的选择性剪接,最终参与 RA 的分子机制,为临床诊断和治疗提供新的研究思路和方向。