Institute of Animal Pathology, Vetsuisse Faculty, University of Bern, Länggassstrasse 122, 3001 Bern, Switzerland.
Dermfocus, Vetsuisse Faculty, University of Bern, 3001 Bern, Switzerland.
Genes (Basel). 2021 Jan 25;12(2):160. doi: 10.3390/genes12020160.
Canine cutaneous epitheliotropic T-cell lymphoma (CETL) and immune-mediated T-cell predominant dermatoses (IMD) share several clinical and histopathological features, but differ substantially in prognosis. The discrimination of ambiguous cases may be challenging, as diagnostic tests are limited and may prove equivocal. This study aimed to investigate transcriptional differences between CETL and IMD, as a basis for further research on discriminating diagnostic biomarkers. We performed 100bp single-end sequencing on RNA extracted from formalin-fixed and paraffin-embedded skin biopsies from dogs with CETL and IMD, respectively. DESeq2 was used for principal component analysis (PCA) and differential gene expression analysis. Genes with significantly different expression were analyzed for enriched pathways using two different tools. The expression of selected genes and their proteins was validated by RT-qPCR and immunohistochemistry. PCA demonstrated the distinct gene expression profiles of CETL and IMD. In total, 503 genes were upregulated, while 4986 were downregulated in CETL compared to IMD. RT-qPCR confirmed the sequencing results for 5/6 selected genes tested, while the protein expression detected by immunohistochemistry was not entirely consistent. Our study revealed transcriptional differences between canine CETL and IMD, with similarities to human cutaneous lymphoma. Differentially expressed genes are potential discriminatory markers, but require further validation on larger sample collections.
犬皮肤上皮性 T 细胞淋巴瘤 (CETL) 和免疫介导的 T 细胞为主的皮肤疾病 (IMD) 具有许多相似的临床和组织病理学特征,但预后却有很大差异。由于诊断测试有限且可能存在不确定性,因此区分这些疾病的不明确病例可能具有挑战性。本研究旨在探讨 CETL 和 IMD 之间的转录差异,作为进一步研究鉴别诊断生物标志物的基础。我们对分别来自患有 CETL 和 IMD 的犬的福尔马林固定和石蜡包埋皮肤活检组织中提取的 RNA 进行了 100bp 单端测序。DESeq2 用于主成分分析 (PCA) 和差异基因表达分析。使用两种不同的工具分析具有显著差异表达的基因的富集途径。通过 RT-qPCR 和免疫组织化学验证选定基因及其蛋白的表达。PCA 显示 CETL 和 IMD 的基因表达谱明显不同。与 IMD 相比,CETL 中有 503 个基因上调,4986 个基因下调。RT-qPCR 验证了 5/6 个选定基因的测序结果,而免疫组织化学检测的蛋白表达并不完全一致。本研究揭示了犬 CETL 和 IMD 之间的转录差异,与人类皮肤淋巴瘤具有相似性。差异表达的基因可能是潜在的鉴别标志物,但需要在更大的样本集中进一步验证。