Restoration of mesenchymal retinal pigmented epithelial cells by TGFβ pathway inhibitors: implications for age-related macular degeneration.
Restoration of mesenchymal retinal pigmented epithelial cells by TGFβ pathway inhibitors: implications for age-related macular degeneration.
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DOI：
10.1186/s13073-015-0183-x
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发表时间：
2015
期刊：
影响因子：
12.3
通讯作者：
Coffey PJ
中科院分区：
文献类型：
--
作者：
Radeke MJ;Radeke CM;Shih YH;Hu J;Bok D;Johnson LV;Coffey PJ
视网膜相关性黄斑变性（AMD）是导致失明的主要原因。大多数视力丧失发生在从存款形成和炎症的疾病转变为新生血管纤维化和/或细胞死亡的疾病之后。在这里，我们研究如何重复的伤口刺激导致基因表达的重大变化和刺激独立的伤口反应在视网膜色素上皮（RPE）细胞，中央AMD的病因细胞类型的永久状态的开始。使用Agilent 44 K全基因组微阵列和RNA-Seq确定人胎儿RPE细胞培养物的转录组宽表达谱作为传代和铺板后时间的函数。使用系统水平的分析，差异表达的基因和途径的利益进行了鉴定，并在建立一个持久的间充质状态的作用进行了评估，使用药理学为基础的实验。使用考虑单层破坏和亚融合培养作为伤口刺激的代表的人胎儿RPE细胞培养模型，我们表明延长的伤口刺激导致融合后间充质表型的终末获得和超过40%的转录组的表达改变。相比之下，在亚汇合处，只有不到5%的表达转录物在重复传代后具有两倍或更大的表达差异。对亚融合培养物中具有传代依赖性表达水平的基因进行蛋白质-蛋白质和途径相互作用分析，揭示了一个158个节点的相互作用组，由两个相互关联的模块组成，其功能与伤口反应和细胞分裂有关。创伤反应基因包括TGFβ通路激活剂：TGFB 1、TGFB 2、INHBA、INHBB、GDF 6、CTGF和THBS 1。值得注意的是，使用受体激酶抑制剂抑制TGFBR 1/ACVR 1B介导的信号传导既能预防又能在很大程度上逆转上皮细胞潜能的E2依赖性丧失;因此将有效寿命延长至少4代。此外，不成比例数量的RPE伤口反应基因改变了新生血管性和地图状AMD中的表达，包括TGFβ途径的关键成员。在RPE细胞中，在长时间的伤口刺激后，TGFβ通路的持续激活驱动了向持久间充质状态的转变。靶向抑制TGFβ信号传导可能是延缓AMD进展和大量产生RPE细胞用于研究和基于细胞的治疗的有效方法。本文的在线版本（doi：10.1186/s13073-015-0183-x）包含补充材料，可供授权用户使用。
Age-related macular degeneration (AMD) is a leading cause of blindness. Most vision loss occurs following the transition from a disease of deposit formation and inflammation to a disease of neovascular fibrosis and/or cell death. Here, we investigate how repeated wound stimulus leads to seminal changes in gene expression and the onset of a perpetual state of stimulus-independent wound response in retinal pigmented epithelial (RPE) cells, a cell-type central to the etiology of AMD. Transcriptome wide expression profiles of human fetal RPE cell cultures as a function of passage and time post-plating were determined using Agilent 44 K whole genome microarrays and RNA-Seq. Using a systems level analysis, differentially expressed genes and pathways of interest were identified and their role in the establishment of a persistent mesenchymal state was assessed using pharmacological-based experiments. Using a human fetal RPE cell culture model that considers monolayer disruption and subconfluent culture as a proxy for wound stimulus, we show that prolonged wound stimulus leads to terminal acquisition of a mesenchymal phenotype post-confluence and altered expression of more than 40 % of the transcriptome. In contrast, at subconfluence fewer than 5 % of expressed transcripts have two-fold or greater expression differences after repeated passage. Protein-protein and pathway interaction analysis of the genes with passage-dependent expression levels in subconfluent cultures reveals a 158-node interactome comprised of two interconnected modules with functions pertaining to wound response and cell division. Among the wound response genes are the TGFβ pathway activators: TGFB1, TGFB2, INHBA, INHBB, GDF6, CTGF, and THBS1. Significantly, inhibition of TGFBR1/ACVR1B mediated signaling using receptor kinase inhibitors both forestalls and largely reverses the passage-dependent loss of epithelial potential; thus extending the effective lifespan by at least four passages. Moreover, a disproportionate number of RPE wound response genes have altered expression in neovascular and geographic AMD, including key members of the TGFβ pathway. In RPE cells the switch to a persistent mesenchymal state following prolonged wound stimulus is driven by lasting activation of the TGFβ pathway. Targeted inhibition of TGFβ signaling may be an effective approach towards retarding AMD progression and producing RPE cells in quantity for research and cell-based therapies. The online version of this article (doi:10.1186/s13073-015-0183-x) contains supplementary material, which is available to authorized users.
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影响因子：
6
作者：
Croze, Roxanne H.;Buchholz, David E.;Clegg, Dennis O.
通讯作者：
Clegg, Dennis O.
影响因子：
4.9
作者：
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3.5
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影响因子：
56.9
作者：
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通讯作者：
Leppert, M