Knockdown of Splicing Complex Protein PCBP2 Reduces Extravillous Trophoblast Differentiation Through Transcript Switching.
Knockdown of Splicing Complex Protein PCBP2 Reduces Extravillous Trophoblast Differentiation Through Transcript Switching.
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剪接复杂蛋白PCBP2的敲低可通过转录作用切换来减少跨滋养细胞的分化。
DOI：
10.3389/fcell.2021.671806
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发表时间：
2021
影响因子：
5.5
通讯作者：
van Dijk M
中科院分区：
文献类型：
--
作者：
Georgiadou D;Boussata S;Keijser R;Janssen DAM;Afink GB;van Dijk M
LINC-HELLP非编码RNA(HELLPAR)的突变与妊娠特异性HELLP综合征的家族形式有关。这些突变对绒毛外滋养细胞(EVT)从增殖状态到侵袭状态的分化有负面影响，并干扰了RNA剪接复合蛋白PCBP1、PCBP2和YBX1与LINC-HELLP的结合。在这项研究中，我们通过体外和体外实验，研究这些蛋白是否参与胎盘形成过程中EVT侵袭的调节。此外，我们还研究了这种调节是否是由于选择性的mRNA剪接。采用HTR-8/SVneo绒毛外滋养层细胞和人早孕胎盘组织，研究siRNA介导的PCBP1、PCBP2和YBX1基因下调对EVT分化的影响。Transwell侵袭试验和增殖试验表明，在PCBP2和YBX1和PCBP1被敲除后，EVT不能分化为侵袭性表型。在胎盘外植体中也观察到了同样的模式，其中PCBP2基因敲除导致显示任何EVT生长的外植体数量减少了约80%。在那些仍然显示出EVT生长的细胞中，增殖的EVT的百分比明显高于转染非靶向对照siRNA的外植体。为了进一步研究PCBP2沉默对EVT的影响，我们对PCBP2基因敲除后的HTR-8/SVneo细胞进行了全转录组测序(RNA-seq)。研究发现，PCBP2基因敲除对基因表达水平的影响很小。相反，PCBP2沉默导致了大量基因剪接的转变，这些基因在细胞组装和组织、细胞功能和维持、细胞生长和增殖以及细胞周期中具有主导功能。EVTS在分化后改变其功能，通过改变它们的细胞组装和通过退出细胞周期来改变它们的增殖活动，从而能够侵入母亲的蜕膜。PCBP2似乎是这些分化机制的重要调节因子，其与LINC-HELLP的结合障碍可能导致HELLP综合征所见的胎盘发育障碍。
Mutations in the LINC-HELLP non-coding RNA (HELLPAR) have been associated with familial forms of the pregnancy-specific HELLP syndrome. These mutations negatively affect extravillous trophoblast (EVT) differentiation from a proliferative to an invasive state and disturb the binding of RNA splicing complex proteins PCBP1, PCBP2, and YBX1 to LINC-HELLP. In this study, by using both in vitro and ex vivo experiments, we investigate if these proteins are involved in the regulation of EVT invasion during placentation. Additionally, we study if this regulation is due to alternative mRNA splicing. HTR-8/SVneo extravillous trophoblasts and human first trimester placental explants were used to investigate the effect of siRNA-mediated downregulation of PCBP1, PCBP2, and YBX1 genes on the differentiation of EVTs. Transwell invasion assays and proliferation assays indicated that upon knockdown of PCBP2 and, to a lesser extent, YBX1 and PCBP1, EVTs fail to differentiate toward an invasive phenotype. The same pattern was observed in placental explants where PCBP2 knockdown led to approximately 80% reduction in the number of explants showing any EVT outgrowth. Of the ones that still did show EVT outgrowth, the percentage of proliferating EVTs was significantly higher compared to explants transfected with non-targeting control siRNAs. To further investigate this effect of PCBP2 silencing on EVTs, we performed whole transcriptome sequencing (RNA-seq) on HTR-8/SVneo cells after PCBP2 knockdown. PCBP2 knockdown was found to have minimal effect on mRNA expression levels. In contrast, PCBP2 silencing led to a switch in splicing for a large number of genes with predominant functions in cellular assembly and organization, cellular function and maintenance, and cellular growth and proliferation and the cell cycle. EVTs, upon differentiation, alter their function to be able to invade the decidua of the mother by changing their cellular assembly and their proliferative activity by exiting the cell cycle. PCBP2 appears to be a paramount regulator of these differentiation mechanisms, where its disturbed binding to LINC-HELLP could contribute to dysfunctional placental development as seen in the HELLP syndrome.
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影响因子：
14.9
作者：
Ji X;Park JW;Bahrami-Samani E;Lin L;Duncan-Lewis C;Pherribo G;Xing Y;Liebhaber SA
通讯作者：
Liebhaber SA
影响因子：
--
作者：
Love, Michael I;Soneson, Charlotte;Patro, Rob
通讯作者：
Patro, Rob
影响因子：
14.9
作者：
Bailey TL;Johnson J;Grant CE;Noble WS
通讯作者：
Noble WS
影响因子：
3.5
作者：
van Dijk, Marie;Visser, Allerdien;Oudejans, Cees B. M.
通讯作者：
Oudejans, Cees B. M.
影响因子：
5.3
作者：
Ruano CSM;Apicella C;Jacques S;Gascoin G;Gaspar C;Miralles F;Méhats C;Vaiman D
通讯作者：
Vaiman D