Mutant U2AF1-induced differential alternative splicing causes an oxidative stress in bone marrow stromal cells
Mutant U2AF1-induced differential alternative splicing causes an oxidative stress in bone marrow stromal cells
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突变体 U2AF1 诱导的差异选择性剪接导致骨髓基质细胞氧化应激
DOI：
10.1177/15353702211010130
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发表时间：
2021-05-25
影响因子：
3.2
通讯作者：
Wang, Jianxiang
中科院分区：
文献类型：
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作者：
Liu, Zhe;Dong, Xuanjia;Wang, Jianxiang
选择性剪接（AS）是基因表达的一个重要调控过程。在骨髓微环境中，AS通过形成不同的重要调控因子亚型，在间充质干细胞命运决定中起着关键作用。作为剪接体因子，U2AF1对pre-mRNA剪接的催化至关重要，其突变可引起不同的As事件。在本研究中，我们通过在小鼠骨髓基质OP9细胞中强制表达突变体U2AF1 (U2AF1S34F)，测定了U2AF1S34F转导的OP9细胞的AS变化，并探讨了其在基质细胞生物学功能中的作用。我们发现U2AF1S34F在OP9细胞中诱导了大量的差异RNA剪接事件。U2AF1S34F导致过氧化氢的生成增加，促进细胞因子和趋化因子的产生。U2AF1S34F转导的OP9细胞也表现出线粒体功能障碍。RNA-seq数据、基因本体（GO）和基因集富集分析显示，响应U2AF1S34F而下调的差异表达基因在过氧化物酶体成分和功能上富集。U2AF1S34F也能引起OP9细胞释放过氧化氢。此外，我们还研究了u2af1s34f诱导的基质细胞氧化应激对造血细胞的影响。小鼠骨髓单核细胞与OP9细胞共培养时，表达OP9细胞的U2AF1S34F诱导造血细胞组蛋白H2AX磷酸化。总之，我们的研究结果表明，突变型u2af1诱导的差异AS事件会引起骨髓基质细胞的氧化应激，并进一步导致造血细胞的DNA损伤和基因组不稳定。
Alternative splicing (AS) is a critical regulatory process of gene expression. In bone marrow microenvironment, AS plays a critical role in mesenchymal stem cells fate determination by forming distinct isoforms of important regulators. As a spliceosome factor, U2AF1 is essential for the catalysis of pre-mRNA splicing, and its mutation can cause differential AS events. In the present study, by forced expression of mutant U2AF1 (U2AF1S34F) in the mouse bone marrow stroma OP9 cells, we determine AS changes in U2AF1S34F transduced OP9 cells and investigate their role in stroma cell biological functions. We find that abundant differential RNA splicing events are induced by U2AF1S34F in OP9 cells. U2AF1S34F causes increased generation of hydrogen peroxide, promotes production of cytokines and chemokines. U2AF1S34F transduced OP9 cells also exhibit dysfunction of mitochondria. RNA-seq data, gene ontology (GO), and gene set enrichment analysis reveal that differentially expressed genes downregulated in response to U2AF1S34F are enriched in peroxisome component and function. U2AF1S34F can also cause release of hydrogen peroxide from OP9 cells. Furthermore, we investigate the influence of U2AF1S34F-induced oxidative stress in stromal cells on hematopoietic cells. When co-culturing mouse bone marrow mononuclear cells with OP9 cells, the U2AF1S34F expressing OP9 cells induce phosphorylation of histone H2AX in hematopoietic cells. Collectively, our results reveal that mutant U2AF1-induced differential AS events cause oxidative stress in bone marrow stromal cells and can further lead to DNA damage and genomic instability in hematopoietic cells.