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Identification of differentially expressed microRNAs in the skin of experimentally sensitized naturally affected atopic beagles by next-generation sequencing.
Immunogenetics ( IF 3.2 ) Pub Date : 2020-03-26 , DOI: 10.1007/s00251-020-01162-w
Domenico Santoro 1 , Antonio Di Loria 2 , Teresa Mirante 3 , Duarte Mendes Oliveira 3 , Carmelo Laudanna 3, 4 , Donatella Malanga 3, 5 , Vincenzo Dattilo 6 , Enrico Iaccino 3 , Rosanna Marsella 1 , Paolo Ciaramella 2
Affiliation  

Canine atopic dermatitis (AD) is a very common inflammatory skin disease, but limited data are available on the genetic characterization (somatic mutations, microarrays, and genome-wide association study (GWAS)) of skin lesions in affected dogs. microRNAs are good biomarkers in inflammatory and neoplastic diseases in people. The aim of this study was to evaluate microRNA expression in the skin of atopic beagles, before and after exposure to Dermatophagoides farinae. Four atopic and four unrelated age-matched healthy beagle dogs were enrolled. Total RNA was extracted from flash-frozen skin biopsies of healthy and atopic dogs. For the atopic dogs, skin biopsies were taken from non-lesional (day 0) and lesional skin (day 28 of weekly environmental challenge with Dermatophagoides farinae). Small RNA libraries were constructed and sequenced. The microRNA sequences were aligned to CanFam3.1 genome. Differential expressed microRNAs were selected on the basis of fold-change and statistical significance (fold-change ≥ 1.5 and p ≤ 0.05 as thresholds. A total of 277 microRNAs were sequenced. One hundred and twenty-one differentially regulated microRNAs were identified between non-lesional and healthy skin. Among these, two were increased amount and 119 were decreased amount. A total of 45 differentially regulated microRNAs between lesional and healthy skin were identified, 44 were decreased amount and one was increased amount. Finally, only two increased amount microRNAs were present in lesional skin when compared with that of non-lesional skin. This is the first study in which dysregulation of microRNAs has been associated with lesional and non-lesional canine AD. Larger studies are needed to understand the role of microRNA in canine AD.

中文翻译:

通过下一代测序鉴定在实验致敏的自然受影响的特应性小猎犬的皮肤中差异表达的microRNA。

犬特应性皮炎(AD)是一种非常常见的炎症性皮肤病,但有关患病犬皮病变的遗传特征(体细胞突变,微阵列和全基因组关联研究(GWAS))的数据有限。microRNA是人类炎症和肿瘤疾病的良好生物标志物。这项研究的目的是评估特应性小猎犬皮肤中暴露于粉红色皮肤癣菌之前和之后的microRNA表达。招募了四只特应性和四只不相关的年龄匹配的健康比格犬。从健康和特应性狗的速冻皮肤活检中提取总RNA。对于特应性犬,从非病变部位(第0天)和病变部位皮肤(每周进行法氏皮肤癣菌环境攻击的第28天)进行皮肤活检。小RNA文库被构建并测序。将微小RNA序列与CanFam3.1基因组比对。根据倍数变化和统计学意义(以倍数变化≥1.5和p≤0.05为阈值)选择差异表达的microRNA。共测序了277个microRNA,在非DNA序列之间鉴定了121个差异调控的microRNA。病变和健康皮肤,其中增加了两个,减少了119个,在病变和健康皮肤之间总共鉴定了45个差异调节的microRNA,减少了44个,增加了一个,最后,只有两个增加的microRNA与非病变皮肤相比,它们存在于病变皮肤中,这是第一项microRNA失调与病变和非病变犬类AD相关的研究。
更新日期:2020-04-21
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