Transcriptome profiling of NIH3T3 cell lines expressing opsin and the P23H opsin mutant identifies candidate drugs for the treatment of retinitis pigmentosa

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• 作者：Yuanyuan Chen^a ; Matthew J. Brooks^b ; Linn Gieser^b ; Anand Swaroop^b ; Krzysztof Palczewski^a ; ^{kxp65@case.edu}
• 关键词：ad ; autosomal dominant ; BP ; biological process ; CHOP ; C/EBP homologous protein ; CRE ; the cAMP-responsive element ; CREB ; the cAMP-responsive element-binding protein ; CREM ; the cAMP-responsive element modulator ; ATF-1 ; activating transcription factor-1 ; DAVID ; the database for annotation visualization and integrated discovery ; DE ; differential expression ; DMEM ; Dulbecco&rsquo ; s modified Eagle&rsquo ; s medium ; ER ; endoplasmic reticulum ; ERAD ; ER-associated protein degradation ; FDA ; Food and Drug Adm
• 刊名：Pharmacological Research
• 出版年：2017
• 出版时间：January 2017
• 年：2017
• 卷：115
• 期：Complete
• 页码：1-13
• 全文大小：4290 K
• 卷排序：115

文摘

Mammalian cells are commonly employed in screening assays to identify active compounds that could potentially affect the progression of different human diseases including retinitis pigmentosa (RP), a class of inherited diseases causing retinal degeneration with compromised vision. Using transcriptome analysis, we compared NIH3T3 cells expressing wildtype (WT) rod opsin with a retinal disease-causing single P23H mutation. Surprisingly, heterologous expression of WT opsin in NIH3T3 cells caused more than a 2-fold change in 783 out of 16,888 protein coding transcripts. The perturbed genes encoded extracellular matrix proteins, growth factors, cytoskeleton proteins, glycoproteins and metalloproteases involved in cell adhesion, morphology and migration. A different set of 347 transcripts was either up- or down-regulated when the P23H mutant opsin was expressed suggesting an altered molecular perturbation compared to WT opsin. Transcriptome perturbations elicited by drug candidates aimed at mitigating the effects of the mutant protein revealed that different drugs targeted distinct molecular pathways that resulted in a similar phenotype selected by a cell-based high-throughput screen. Thus, transcriptome profiling can provide essential information about the therapeutic potential of a candidate drug to restore normal gene expression in pathological conditions.