Severe hypoxia exerts parallel and cell-specific regulation of gene expression and alternative splicing in human mesenchymal stem cells.
Severe hypoxia exerts parallel and cell-specific regulation of gene expression and alternative splicing in human mesenchymal stem cells.
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严重缺氧对人间充质干细胞中的基因表达和选择性剪接产生平行且细胞特异性的调节
DOI：
10.1186/1471-2164-15-303
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发表时间：
2014-04-23
期刊：
影响因子：
4.4
通讯作者：
Wang J
中科院分区：
文献类型：
--
作者：
Hu X;Wu R;Shehadeh LA;Zhou Q;Jiang C;Huang X;Zhang L;Gao F;Liu X;Yu H;Webster KA;Wang J
骨髓内皮提供了一个特殊的缺氧生态位，可能有助于保持间充质干细胞（MSCs）的完整性、多能性、寿命和干性。为了探索这种生态位的分子遗传学后果，我们将人(h)间充质干细胞置于po2为4 mmHg的环境中，并通过基因组-外显子微阵列和RT-qPCR分析了整体基因表达和选择性剪接（AS），并通过western blot和免疫染色分析了表型。结果在446个>2.5倍差异调控基因中，下调基因数量比上调基因数量多243:203。外显子分析显示，53个基因中有60个低氧调节的AS事件，剪接指数（SI）为bbb1.0，高SI与转录调节程度相关。对一项公开的人脐静脉内皮细胞（HUVECs） AS研究的平行分析表明，在hMSCs和HUVECs中只有11个共同调控的基因和17个共同的差异剪接基因具有很强的细胞特异性成分。在两种细胞类型中，只有3个基因在基因（>2.0）和AS水平上对缺氧反应存在差异。功能定位揭示了分化、细胞外基质、中间丝和代谢标记基因复杂调控的独特基因表达谱。抗氧化基因、横纹肌基因和胰岛素/IGF-1信号中间体下调。12个酸性角蛋白中有9个与其他上皮和细胞粘附标志物协同诱导，表明部分间充质向上皮过渡。我们得出结论，严重缺氧赋予hMSCs静默表型，这反映在转录组谱和剪接体作用的基因特异性变化上。结果表明，与中度缺氧相比，重度缺氧对MSCs的基因调控模式有明显不同。这是第一个报道缺氧对AS在干细胞/祖细胞中的调节的研究，也是第一个在缺氧诱导的静止不动状态下MSC的分子遗传学表征。
BackgroundThe endosteum of the bone marrow provides a specialized hypoxic niche that may serve to preserve the integrity, pluripotency, longevity and stemness of resident mesenchymal stem cells (MSCs). To explore the molecular genetic consequences of such a niche we subjected human (h) MSCs to a pO2of 4 mmHg and analyzed global gene expression and alternative splicing (AS) by genome-exon microarray and RT-qPCR, and phenotype by western blot and immunostaining.ResultsOut of 446 genes differentially regulated by >2.5-fold, down-regulated genes outnumbered up-regulated genes by 243:203. Exon analyses revealed 60 hypoxia-regulated AS events with splice indices (SI) >1.0 from 53 genes and a correlation between high SI and degree of transcript regulation. Parallel analyses of a publicly available AS study on human umbilical vein endothelial cells (HUVECs) showed that there was a strong cell-specific component with only 11 genes commonly regulated in hMSCs and HUVECs and 17 common differentially spliced genes. Only 3 genes were differentially responsive to hypoxia at the gene (>2.0) and AS levels in both cell types. Functional assignments revealed unique profiles of gene expression with complex regulation of differentiation, extracellular matrix, intermediate filament and metabolic marker genes. Antioxidant genes, striated muscle genes and insulin/IGF-1 signaling intermediates were down-regulated. There was a coordinate induction of 9 out of 12 acidic keratins that along with other epithelial and cell adhesion markers implies a partial mesenchymal to epithelial transition.ConclusionsWe conclude that severe hypoxia confers a quiescent phenotype in hMSCs that is reflected by both the transcriptome profile and gene-specific changes of splicosome actions. The results reveal that severe hypoxia imposes markedly different patterns of gene regulation of MSCs compared with more moderate hypoxia. This is the first study to report hypoxia-regulation of AS in stem/progenitor cells and the first molecular genetic characterization of MSC in a hypoxia-induced quiescent immobile state.
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影响因子：
37.8
作者：
Hu S;Huang M;Li Z;Jia F;Ghosh Z;Lijkwan MA;Fasanaro P;Sun N;Wang X;Martelli F;Robbins RC;Wu JC
通讯作者：
Wu JC
影响因子：
10.5
作者：
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通讯作者：
Manley, James L.
影响因子：
8
作者：
Anderson, CJ;Hoare, SF;Keith, WN
通讯作者：
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影响因子：
3.7
作者：
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通讯作者：
Prockop DJ
影响因子：
2.8
作者：
Hung, Shun-Pei;Ho, Jennifer H.;Lee, Oscar K.
通讯作者：
Lee, Oscar K.