Singh Jasdeep, Dhanoa Jasdeep Kaur, Choudhary Ratan K, Singh Amarjit, Sethi Ram Saran, Kaur Simarjeet, Mukhopadhyay Chandra Sekhar
College of Animal Biotechnology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab 141004 India.
Present address: University Institute of Biotechnology (UIBT), Chandigarh University, Mohali, Punjab 140413 India.
ExRNA. 2020;2(1):8. doi: 10.1186/s41544-020-00049-y. Epub 2020 May 22.
MicroRNAs play key roles in host-pathogen-interactions and disease pathogenesis. Our aim was to characterize the differentially expressed miRNAs in the blood cells of diseased (Brucellosis-positive, Johne's disease-positive) and healthy- water buffaloes. The pooled small-RNA samples of each group were sequenced on Ion Torrent Personal Genome Machine (PGM) sequencer and the data were analyzed for differential expression.
Here we identified 274 known miRNAs with bovine homologs and 36 novel mature-star miRNAs from the sequnces of small RNA libraries. Overall 195 miRNAs were common to all the three groups. Certain miRNAs such as bta-miR-21-5p, -26a, -29a/b, -30d - 103, - 140, - 150, - 191, - 374, - 1434-5p,-1260b, - 2484 and let-7 members were abundantly expressed in diseased groups. Bta-miR-1434-5p, - 188, -200c were up-regulated (> 1.5 folds) while bta-miR-27a-5p, -34b and -2285x were down-regulated (> 100 folds) in Brucellosis group. In Johne's Disease group, only 3 miRNAs (bta-miR-1434-5p, - 2340 and - 2484) were up-regulated (> 1.5 folds). The functional classification of miRNA target genes into gene ontology (GO) terms indicated their involvement in innate immunity and cellular process of disease pathogenesis. Expression profile of four differentially expressed miRNAs (bta-miR-9-5p, - 677, - 331-3p and - 2440) and eight predicted target-genes were validated through reverse transcriptase qPCR.
This study provides a valuable frame of reference for elucidation of regulatory roles of miRNAs associated with disease pathogenesis in water buffaloes as well as identification of miRNA biomarkers for disease diagnosis and treatment.
微小RNA在宿主-病原体相互作用和疾病发病机制中发挥关键作用。我们的目的是鉴定患病(布鲁氏菌病阳性、副结核阳性)和健康水牛血细胞中差异表达的微小RNA。对每组混合的小RNA样本在Ion Torrent个人基因组测序仪(PGM)上进行测序,并对数据进行差异表达分析。
我们从小RNA文库序列中鉴定出274个具有牛同源物的已知微小RNA和36个新的成熟-星型微小RNA。三组共有的微小RNA有195个。某些微小RNA,如bta-miR-21-5p、-26a、-29a/b、-30d、-103、-140、-150、-191、-374、-1434-5p、-1260b、-2484以及let-7成员在患病组中大量表达。在布鲁氏菌病组中,bta-miR-1434-5p、-188、-200c上调(>1.5倍),而bta-miR-27a-5p、-34b和-2285x下调(>100倍)。在副结核组中,只有3个微小RNA(bta-miR-1434-5p、-2340和-2484)上调(>1.5倍)。将微小RNA靶基因的功能分类为基因本体(GO)术语表明它们参与天然免疫和疾病发病机制的细胞过程。通过逆转录定量PCR验证了4个差异表达微小RNA(bta-miR-9-5p、-677、-331-3p和-2440)和8个预测靶基因的表达谱。
本研究为阐明微小RNA在水牛疾病发病机制中的调控作用以及鉴定用于疾病诊断和治疗的微小RNA生物标志物提供了有价值的参考框架。