Insulin-like growth factor 2 mRNA-binding protein 2-regulated alternative splicing of nuclear factor 1 C-type causes excessive granulosa cell proliferation in polycystic ovary syndrome.
Insulin-like growth factor 2 mRNA-binding protein 2-regulated alternative splicing of nuclear factor 1 C-type causes excessive granulosa cell proliferation in polycystic ovary syndrome.
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IGF2BP2 调节的 NFIC 选择性剪接导致 PCOS 中颗粒细胞过度增殖
DOI：
10.1111/cpr.13216
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发表时间：
2022-04
影响因子：
8.5
通讯作者：
Yang, Xiaokui
中科院分区：
文献类型：
--
作者：
Zhao, Feiyan;Wu, Liang;Wang, Qin;Zhao, Xuehan;Chen, Tong;Yin, Chenghong;Yan, Long;Yang, Xiaokui
多囊卵巢综合征（PCOS）是一种常见的生殖内分泌疾病。胰岛素样生长因子2 mRNA结合蛋白2（IGF 2BP 2）作为HMGA 2靶基因促进颗粒细胞（GC）增殖。然而，IGF 2BP 2是否作为RNA结合蛋白（RBP）参与PCOS的发病机制尚不清楚。在这项研究中，我们的目的是阐明IGF 2BP 2相互作用的转录本，全局转录组以及GC中的选择性剪接，以最终揭示PCOS发病的潜在机制。采用RT-qPCR和western blot检测PCOS患者GCs中IGF 2BP 2的表达。我们通过RNA免疫沉淀测序（RIP‐seq）和RNA测序（RNA‐seq）捕获IGF 2BP 2相互作用转录物、全局转录组以及可变剪接。将用IGF 2BP 2过表达质粒和核因子1 C型（NFIC）siRNA转染的KGN细胞用于CCK-8、EdU和TUNEL试验。IGF 2BP 2在PCOS患者的GC中呈高表达。作为RBP，它优先与具有GGAC基序和新发现的GAAG基序的mRNA的3′和5′ UTR结合。IGF 2BP 2的过表达改变了KGN细胞的转录组谱。IGF 2BP 2通过抑制NFIC的外显子跳跃事件，调节选择性剪接事件，促进细胞增殖。总之，我们证明IGF 2BP 2通过调节PCOS中NFIC的选择性剪接来促进GC增殖。这一发现有助于更好地理解IGF 2BP 2在PCOS发病机制中的作用。多囊卵巢综合征（PCOS）是一种常见的生殖内分泌疾病，影响约10%的育龄妇女。胰岛素样生长因子2 mRNA结合蛋白2（IGF 2BP 2）是一种RNA结合蛋白（RBP），通过结合RNA转录物调节一系列靶基因。然而，IGF 2BP 2是否参与PCOS的发病机制尚不清楚。在这项研究中，我们检测了IGF 2BP 2在PCOS患者颗粒细胞（GCs）中的表达，并在永生化的人GCs（KGN细胞）中捕获IGF 2BP 2相互作用的转录物，全局转录组以及RIP-seq和RNA-seq的选择性剪接。我们的研究结果表明IGF 2BP 2在PCOS患者的GC中过表达。作为RBP，它优先与具有GGAC基序和新发现的GAAG基序的mRNA的3′和5′ UTR结合。此外，IGF 2BP 2的过表达可通过调节KGN细胞中NFIC的选择性剪接来促进细胞增殖。总之，我们提供了一个转录组范围的分析，证明了IGF 2BP 2在PCOS发病机制中的作用。本研究的结果将有助于我们从IGF 2BP 2介导的选择性剪接的角度更好地了解PCOS的病因，并为未来PCOS的个人治疗指导提供机会。
Polycystic ovary syndrome (PCOS) is a common reproductive endocrine disorder. Insulin‐like growth factor 2 mRNA‐binding protein 2 (IGF2BP2) serves as an HMGA2 target gene to promote the proliferation of granulosa cells (GCs). However, it is still unclear whether IGF2BP2 participates in the pathogenesis of PCOS as RNA binding protein (RBP). In this study, we aimed to elucidate IGF2BP2‐interacting transcripts, global transcriptome together with alternative splicing in GCs to eventually uncover potential mechanisms of PCOS pathogenesis. The expression of IGF2BP2 in GCs from PCOS patients was detected using quantitative reverse transcription PCR (RT‐qPCR) and western blot. We captured IGF2BP2‐interacting transcripts, global transcriptome together with alternative splicing by RNA immunoprecipitation sequencing (RIP‐seq) and RNA sequencing (RNA‐seq). KGN cells transfected with IGF2BP2 overexpressing plasmids and nuclear factor 1 C‐type (NFIC) siRNAs, were applied to CCK‐8, EdU and TUNEL assays. IGF2BP2 was highly expressed in GCs from PCOS patients. As an RBP, it preferentially bound to the 3′and 5′UTRs of mRNAs with GGAC motif and a newly found GAAG motif. The overexpression of IGF2BP2 changed the transcriptome profile of KGN cells. IGF2BP2 functioned to regulate alternative splicing events and promote cell proliferation through inhibiting exon skipping events of NFIC. In conclusion, we demonstrated that IGF2BP2 promotes GC proliferation via regulating alternative splicing of NFIC in PCOS. The findings help to better understand the roles of IGF2BP2 in the pathogenesis of PCOS. Polycystic ovary syndrome (PCOS) is a common reproductive endocrine disorder, affecting approximately 10% of women of reproductive age. Insulin‐like growth factor 2 mRNA‐binding protein 2 (IGF2BP2) reportedly functions as an RNA‐binding proteins (RBPs) that regulates a series of target genes by binding RNA transcripts. However, it is still unclear whether IGF2BP2 participates in pathogenesis of PCOS. In this study, we detected expression of IGF2BP2 in granulosa cells (GCs) from PCOS patients and captured IGF2BP2‐interacting transcripts, global transcriptome together with alternative splicing by RIP‐seq and RNA‐seq in immortalized human GCs (KGN cells). Our results show that IGF2BP2 is overexpressed in GCs from PCOS patients. As an RBP, it preferentially binds to the 3′and 5′UTRs of mRNAs with GGAC motif and a newly found GAAG motif. Moreover, overexpression of IGF2BP2 can promote cell proliferation through regulating alternative splicing of NFIC in KGN cells. In conclusion, we provide a transcriptome‐wide analysis that demonstrates the role played by IGF2BP2 in the pathogenesis of PCOS. The findings of this study will help us to better understand the aetiology of PCOS from the perspective of IGF2BP2 mediated alternative splicing as well as suggest opportunities for future personal therapeutic guidance for PCOS.
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DOI：
10.1038/nrm.2017.27
发表时间：
2017-07
期刊：
Nature reviews. Molecular cell biology
影响因子：
--
作者：
Baralle FE;Giudice J
通讯作者：
Giudice J
影响因子：
3.8
作者：
Ruchat, Stephanie-May;Elks, Cathy E.;Perusse, Louis
通讯作者：
Perusse, Louis
影响因子：
3.7
作者：
Liang, Xin;Gao, Jiangang;Wu, Changli
通讯作者：
Wu, Changli
影响因子：
6.1
作者：
Fauser, BCJM;Chang, J;Lobo, R
通讯作者：
Lobo, R
影响因子：
9
作者：
Chen F;Wang Q;Yu X;Yang N;Wang Y;Zeng Y;Zheng Z;Zhou F;Zhou Y
通讯作者：
Zhou Y