Regulation of WNT Signaling by VSX2 During Optic Vesicle Patterning in Human Induced Pluripotent Stem Cells.
Regulation of WNT Signaling by VSX2 During Optic Vesicle Patterning in Human Induced Pluripotent Stem Cells.
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DOI：
10.1002/stem.2414
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发表时间：
2016-11
期刊：
影响因子：
5.2
通讯作者：
Gamm, David M.
中科院分区：
文献类型：
--
作者：
Capowski, Elizabeth E.;Wright, Lynda S.;Liang, Kun;Phillips, M. Joseph;Wallace, Kyle;Petelinsek, Anna;Hagstrom, Anna;Pinilla, Isabel;Borys, Katarzyna;Lien, Jessica;Min, Jee Hong;Keles, Sunduz;Thomson, James A.;Gamm, David M.
尽管视觉系统同源框2(VSX2)在视网膜形成早期对维持神经视网膜(NR)的命运起着广泛而关键的作用，但几乎没有发现它的基因靶点。我们对来自人iPS细胞(HiPSCs)的早期视泡泡样结构(OV)进行了VSX2 CHIP-SEQ和CHIP-PCR检测，突出了WNT通路基因是VSX2的直接调控靶点。对来自VSX2功能性零突变患者的HiPSC-OV的早期NR模式的检查显示，与对照HiPSC-OV相比，WNT途径成分和视网膜色素上皮(RPE)标记错误表达和上调。此外，药物抑制WNT信号挽救了早期突变的表型，而在对照HiPSC-OVS中WNT信号的增强拯救了突变体的表型。这些发现揭示了VSX2作为WNT信号调节因子的重要作用，并提示VSX2可能通过直接抑制WNT信号通路成分而以牺牲RPE为代价来维持NR的同一性。视觉系统同源框2(VSX2)是参与视网膜神经发育的关键转录因子。然而，令人惊讶的是，除了小眼炎诱导转录因子(MITF)之外，已知的VSX2基因调控靶点很少，MITF是一种参与视网膜色素上皮(RPE)发育的蛋白质。我们使用患者特有的人类多能干细胞携带VSX2突变，该突变取消了DNA结合活性(R200Q)，结合药物操作和染色质免疫沉淀，展示了VSX2和WNT信号基因之间的一种新的相互作用。在VSX2R200Q培养中，VSX2与WNT信号基因的结合被取消，视泡细胞异常地采用RPE命运代替神经视网膜。这种效应可以通过抑制WNT信号部分克服，或通过促进WNT信号在野生型细胞中诱导。这项研究还提出了第一份无偏见的VSX2靶标清单，从而为识别参与这一生物过程的其他基因和机制提供了资源。
Few gene targets of Visual System Homeobox 2 (VSX2) have been identified despite its broad and critical role in the maintenance of neural retina (NR) fate during early retinogenesis. We performed VSX2 ChIP-seq and ChIP-PCR assays on early stage optic vesicle-like structures (OVs) derived from human iPS cells (hiPSCs), which highlighted WNT pathway genes as direct regulatory targets of VSX2. Examination of early NR patterning in hiPSC-OVs from a patient with a functional null mutation in VSX2 revealed mis-expression and upregulation of WNT pathway components and retinal pigmented epithelium (RPE) markers in comparison to control hiPSC-OVs. Furthermore, pharmacological inhibition of WNT signaling rescued the early mutant phenotype, whereas augmentation of WNT signaling in control hiPSC-OVs phenocopied the mutant. These findings reveal an important role for VSX2 as a regulator of WNT signaling and suggest that VSX2 may act to maintain NR identity at the expense of RPE in part by direct repression of WNT pathway constituents. Visual systems homeobox 2 (VSX2) is a key transcription factor involved in neural retinal development. However, surprisingly few gene regulatory targets are known for VSX2 beyond Microphthalmia induced transcription factor (MITF), a protein involved in the development of retinal pigmented epithelium (RPE). We used patient-specific human pluripotent stem cells carrying a mutation in VSX2 that abolishes DNA binding activity (R200Q), combined with pharmacological manipulations and chromatin immunoprecipitation, to demonstrate a novel interaction between VSX2 and WNT signaling genes. In VSX2R200Q cultures, VSX2 binding to WNT signaling genes is abolished and optic vesicle cells aberrantly adopt an RPE fate in lieu of neural retina. This effect that can be partially overcome by inhibition of WNT signaling, or induced in wild type cells through promotion of WNT signaling. This study also presents the first unbiased list of VSX2 targets, thus providing a resource for identification of other genes and mechanisms involved in this biological process.
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影响因子：
2.7
作者：
Fujimura, Naoko;Taketo, Makoto M.;Kozmik, Zbynek
通讯作者：
Kozmik, Zbynek
影响因子：
3.7
作者：
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Heinz S;Benner C;Spann N;Bertolino E;Lin YC;Laslo P;Cheng JX;Murre C;Singh H;Glass CK
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Glass CK
DOI：
10.1073/pnas.0601990103
发表时间：
2006-08-22
影响因子：
11.1
作者：
Lamba, Deepak A.;Karl, Mike O.;Reh, Thomas A.
通讯作者：
Reh, Thomas A.
影响因子：
12.3
作者：
Langmead B;Trapnell C;Pop M;Salzberg SL
通讯作者：
Salzberg SL