Li Bingshan, Tsoi Lam C, Swindell William R, Gudjonsson Johann E, Tejasvi Trilokraj, Johnston Andrew, Ding Jun, Stuart Philip E, Xing Xianying, Kochkodan James J, Voorhees John J, Kang Hyun M, Nair Rajan P, Abecasis Goncalo R, Elder James T
Department of Molecular Physiology and Biophysics, Center for Human Genetics Research, Vanderbilt University, Nashville, Tennessee, USA; Department of Biostatistics, University of Michigan, Ann Arbor, Michigan, USA.
Department of Biostatistics, University of Michigan, Ann Arbor, Michigan, USA.
J Invest Dermatol. 2014 Jul;134(7):1828-1838. doi: 10.1038/jid.2014.28. Epub 2014 Jan 17.
To increase our understanding of psoriasis, we used high-throughput complementary DNA sequencing (RNA-seq) to assay the transcriptomes of lesional psoriatic and normal skin. We sequenced polyadenylated RNA-derived complementary DNAs from 92 psoriatic and 82 normal punch biopsies, generating an average of ∼38 million single-end 80-bp reads per sample. Comparison of 42 samples examined by both RNA-seq and microarray revealed marked differences in sensitivity, with transcripts identified only by RNA-seq having much lower expression than those also identified by microarray. RNA-seq identified many more differentially expressed transcripts enriched in immune system processes. Weighted gene coexpression network analysis (WGCNA) revealed multiple modules of coordinately expressed epidermal differentiation genes, overlapping significantly with genes regulated by the long noncoding RNA TINCR, its target gene, staufen-1 (STAU1), the p63 target gene ZNF750, and its target KLF4. Other coordinately expressed modules were enriched for lymphoid and/or myeloid signature transcripts and genes induced by IL-17 in keratinocytes. Dermally expressed genes were significantly downregulated in psoriatic biopsies, most likely because of expansion of the epidermal compartment. These results show the power of WGCNA to elucidate gene regulatory circuits in psoriasis, and emphasize the influence of tissue architecture in both differential expression and coexpression analysis.
为了加深我们对银屑病的理解,我们使用高通量互补DNA测序(RNA测序)来分析银屑病皮损和正常皮肤的转录组。我们对来自92例银屑病和82例正常皮肤打孔活检样本的聚腺苷酸化RNA衍生的互补DNA进行测序,每个样本平均产生约3800万个单端80碱基读取。对42个同时通过RNA测序和微阵列检测的样本进行比较,发现灵敏度存在显著差异,仅通过RNA测序鉴定出的转录本表达水平远低于那些也通过微阵列鉴定出的转录本。RNA测序鉴定出更多在免疫系统过程中富集的差异表达转录本。加权基因共表达网络分析(WGCNA)揭示了多个协调表达的表皮分化基因模块,与由长链非编码RNA TINCR、其靶基因Staufen-1(STAU1)、p63靶基因ZNF750及其靶基因KLF4调控的基因有显著重叠。其他协调表达的模块富含淋巴细胞和/或髓细胞特征转录本以及角质形成细胞中由IL-17诱导的基因。在银屑病活检样本中,真皮表达基因显著下调,最可能的原因是表皮区域的扩张。这些结果显示了WGCNA在阐明银屑病基因调控回路方面的作用,并强调了组织结构在差异表达和共表达分析中的影响。