Takada Akane, Kamatari Yuji O, Shimizu Kaori, Okada Ayaka, Inoshima Yasuo
Laboratory of Food and Environmental Hygiene, Cooperative Department of Veterinary Medicine, Faculty of Applied Biological Sciences, Gifu University, 1-1 Yanagido, Gifu 501-1193, Japan.
Institute of Glyco-Core Research (iGCORE), Gifu University, 1-1 Yanagido, Gifu 501-1193, Japan.
Microorganisms. 2023 Aug 28;11(9):2173. doi: 10.3390/microorganisms11092173.
Enzootic bovine leukosis (EBL) is a B-cell lymphosarcoma caused by the bovine leukemia virus (BLV). While most infected cattle show no clinical signs, approximately 30% of infected cattle develop persistent lymphocytosis (PL), and a small percentage may develop EBL. Currently, there is no method for predicting the possibility of EBL onset. In this study, we analyzed the microRNAs (miRNAs) encapsulated in small extracellular vesicles (sEVs) in the blood to explore the biomarkers of EBL. To identify candidate biomarkers, blood samples were collected from three BLV-uninfected and three EBL cattle. Total RNA was extracted from filtered serum and used for microarray analysis. Due to their association with cancer in human orthologs, we selected three miRNAs as candidate biomarkers, bta-miR-17-5p, bta-miR-24-3p, and bta-miR-210, which were more than twice as abundant in EBL cattle than in BLV-uninfected cattle. Quantitative real-time polymerase chain reaction (qPCR) using serum RNAs from six cattle used for the microarray analysis was carried out for the detection of the three selected miRNAs. Additionally, bta-miR-92a, whose ortholog has been associated with cancer in humans, was also examined by qPCR. bta-miR-17-5p, bta-miR-24-3p, and bta-miR-92a, were successfully detected, but bta-miR-210 was not. To further evaluate the utility of these three miRNAs as biomarkers, new blood samples were collected from 31 BLV-uninfected and 30 EBL cattle. The levels of bta-miR-17-5p, bta-miR-24-3p, and bta-miR-92a, were significantly higher in EBL cattle than in BLV-uninfected cattle. These results suggest that increased levels of bta-miR-17-5p, bta-miR-24-3p, and bta-miR-92a in the blood could be used as biomarkers for EBL. This study may contribute to the control of BLV infections and develop a prediction method of EBL onset.
地方流行性牛白血病（EBL）是一种由牛白血病病毒（BLV）引起的B细胞淋巴瘤。虽然大多数感染牛没有临床症状，但约30%的感染牛会出现持续性淋巴细胞增多症（PL），还有一小部分可能会发展为EBL。目前，尚无预测EBL发病可能性的方法。在本研究中，我们分析了血液中小细胞外囊泡（sEVs）中包裹的微小RNA（miRNAs），以探索EBL的生物标志物。为了鉴定候选生物标志物，从3头未感染BLV的牛和3头EBL牛采集血样。从过滤后的血清中提取总RNA并用于微阵列分析。由于它们在人类直系同源物中与癌症相关，我们选择了3种miRNAs作为候选生物标志物，即bta-miR-17-5p、bta-miR-
