高、低抗蜱牛的差异表达细胞外囊泡、外泌体和非外泌体 miRNA 图谱。

Differentially Expressed Extracellular Vesicle, Exosome and Non-Exosome miRNA Profile in High and Low Tick-Resistant Beef Cattle.

机构信息

Centre for Children's Health Research, School of Biomedical Sciences, Faculty of Health, The Queensland University of Technology, Brisbane, QLD, Australia.

Nindooinbah Pastoral Company, Nindooinbah, QLD, Australia.

出版信息

Front Cell Infect Microbiol. 2021 Dec 17;11:780424. doi: 10.3389/fcimb.2021.780424. eCollection 2021.

DOI:10.3389/fcimb.2021.780424

PMID:34976862

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8718928/

Abstract

Heavy tick burden on beef cattle account for huge economic losses globally, with an estimated value of US$22-30 billion per annum. In Australia, ticks cost the northern beef industry approximately A$170-200 million. Methods to evaluate and predict tick resistance would therefore be of great value to the global cattle trade. Exosomes (EX) are small extracellular vesicles (EVs) of ~30-150nm diameter and have gained popularity for their diagnostic and prognostic potential. EX contain, among other biomolecules, various types of RNA including micro-RNA (miRNA) and long noncoding RNA (lncRNA). MiRNA specifically have been validated as therapeutic biomarkers as they perform regulatory functions at the post-transcriptional level and are differentially expressed between divergent groups. The objective of the present study was to evaluate the miRNA profiles of EV and fractionated exosomal samples of high and low tick-resistant beef cattle to highlight potential miRNA biomarkers of tick resistance. Cows ( = 3/group) were classified into high or low tick resistant groups according to a novel scoring system. EVs and EX were isolated and fractionated from the blood plasma of high and low tick resistant cattle using established isolation and enrichment protocols. The resultant EX and non-EX samples were processed for next generation miRNA sequencing. Offspring of the cows in each high and low tick resistant group underwent the same processing for blood plasma EX, non-EX and miRNA analysis to evaluate the heritability of miRNA associated with tick resistance. A total of 2631 miRNAs were identified in EX and non-EX fractionated samples from high and low tick-resistant beef cattle. MiR-449a was highly expressed in maternal high tick-resistant EX samples. Of these, 174 were novel miRNAs, and 10 were differentially expressed (DE) (FDR < 0.05). These 10 DE miRNAs were also present in EVs, and three miRNAs were highly expressed: miR-2419-3p, miR-7861-3p and miR-2372-5p. Although 196 novel miRNAs were identified in fractionated samples of offspring, no miRNA were differentially expressed in these animals.

摘要

牛身上的蜱虫负担沉重，给全球造成了巨大的经济损失，估计每年的损失价值为 220 亿至 300 亿美元。在澳大利亚，蜱虫给北部养牛业造成的损失约为 1.7 亿至 2 亿澳元。因此，评估和预测蜱虫抗药性的方法将对全球牛贸易具有重要价值。外泌体（EX）是直径约 30-150nm 的小细胞外囊泡（EVs），由于其具有诊断和预后潜力而受到广泛关注。EX 中含有各种生物分子，包括 micro-RNA (miRNA) 和长非编码 RNA (lncRNA) 等。miRNA 已被验证为治疗性生物标志物，因为它们在转录后水平发挥调节功能，并且在不同群体之间存在差异表达。本研究的目的是评估高、低蜱抗性肉牛血液中 EV 和分离的 EX 样本的 miRNA 图谱，以突出蜱抗性的潜在 miRNA 生物标志物。根据一种新的评分系统，将奶牛（每组 3 头）分为高或低蜱抗性组。使用已建立的分离和富集方案从高、低蜱抗性牛的血液血浆中分离和分离 EV 和 EX。从高、低蜱抗性牛的血液血浆中分离和分离 EV 和 EX。分离的 EX 和非 EX 样本用于下一代 miRNA 测序。每组高、低蜱抗性牛的后代也接受了相同的血液 EX、非 EX 和 miRNA 分析处理，以评估与蜱抗性相关的 miRNA 的遗传性。从高、低蜱抗性肉牛的 EX 和非 EX 分离样本中鉴定出 2631 个 miRNA。母系高蜱抗性 EX 样本中 miR-449a 高度表达。其中，174 个是新的 miRNA，10 个是差异表达（FDR < 0.05）。这些 10 个差异表达的 miRNA 也存在于 EV 中，其中 3 个 miRNA 高度表达：miR-2419-3p、miR-7861-3p 和 miR-2372-5p。尽管在分离的后代样本中鉴定出 196 个新的 miRNA，但这些动物中没有 miRNA 表达差异。