Single-nucleotide-polymorphism-adjacent super enhancer network mediates enhanced osteogenic differentiation of mesenchymal stem cells in ankylosing spondylitis [RNA-seq]

In order to recapitulated the primary mechanism of the pathologically enhanced osteogenesis of mesenchymal stem cells from ankylosing spondylitis patients (ASMSCs) over MSCs from healthy donor, we performed multiomic high-throughput sequencing. We analysed the H3K27ac ChIP-seq data according to ROSE algorithm, and identified super enhancers (SEs) in ASMSCs and HDMSCs. The ASMSC-unique SEs (ASUSEs) are associated with osteogenic differentiation and AS pathogenesis. By integrated analysis of H3K27ac ChIP-seq, ankylosing spondylitis (AS) SNPs and RNA-seq data, we discovered the transcription network regulated by AS SNP-adjacent super enhancers (SASEs) during the enhanced osteogenic differentiation of MSCs from AS patients (ASMSCs), which helped us gain insight into the crutial mechanism of AS pathological osteogenesis. And based on the inhibition effect of SE inhibitor JQ1 on the enhanced osteogenic differentiation of ASMSCs, we proposed that SEs may be attractive targets to treat AS pathological osteogenesis. Overall design: Examination of the mRNA profiles in ASMSCs and HDMSCs during osteogenic differentiation

为复现强直性脊柱炎（ankylosing spondylitis, AS）患者来源间充质干细胞（mesenchymal stem cells, MSCs，即ASMSCs）相较于健康供体来源间充质干细胞（HDMSCs）的病理性增强成骨核心机制，本研究开展了多组学高通量测序。我们依据ROSE算法分析H3K27ac染色质免疫共沉淀测序（ChIP-seq）数据，在ASMSCs与HDMSCs中鉴定出超级增强子（super enhancers, SEs）。其中，AS特异性超级增强子（ASUSEs）与成骨分化及AS发病机制密切相关。通过整合分析H3K27ac ChIP-seq、AS单核苷酸多态性（single nucleotide polymorphisms, SNPs）及RNA测序（RNA-seq）数据，我们揭示了在ASMSCs增强成骨分化过程中，由AS单核苷酸多态性邻近超级增强子调控的转录网络，进而加深了对AS病理性成骨关键机制的理解。基于超级增强子抑制剂JQ1对ASMSCs增强成骨分化的抑制作用，我们提出超级增强子或可成为治疗AS病理性成骨的潜在靶点。整体实验设计：检测成骨分化过程中ASMSCs与HDMSCs的mRNA表达谱。