Sun Chengsong, Luo Fang, You Yanmin, Gu Mengjie, Yang Wenbin, Yi Cun, Zhang Wei, Feng Zheng, Wang Jipeng, Hu Wei
Department of Infectious Diseases, Huashan Hospital, State Key Laboratory of Genetic Engineering, Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering, Ministry of Education Key Laboratory of Contemporary Anthropology, School of Life Sciences, Fudan University, No. 2005 Songhu Road, Yangpu District, Shanghai 200438, China; Anhui Provincial Institute of Parasitic Diseases, No. 12560 Fanhua Avenue, Shushan District, Hefei 230601, Anhui Province, China.
Department of Infectious Diseases, Huashan Hospital, State Key Laboratory of Genetic Engineering, Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering, Ministry of Education Key Laboratory of Contemporary Anthropology, School of Life Sciences, Fudan University, No. 2005 Songhu Road, Yangpu District, Shanghai 200438, China.
Int J Parasitol. 2023 Oct;53(11-12):637-649. doi: 10.1016/j.ijpara.2023.03.007. Epub 2023 Jun 22.
Eggs laid by mature female schistosomes are primarily responsible for the pathogenesis of schistosomiasis and critical for transmission. Consequently, elucidating the mechanism of sexual maturation as well as egg production may lead to new strategies for the control of schistosomiasis. MicroRNAs (miRNAs) are involved in multiple biological processes including reproduction in many organisms, yet their roles have not been well characterized in schistosomes. Here, we investigated microRNA-1 (miR-1), which was downregulated gradually in both male and female Schistosoma japonicum after they reached sexually maturity. The expression of miR-1, as shown with quantitative reverse transcription PCR (qRT-PCR), was lower in the reproductive organs of adult females compared with the somatic tissues. Overexpression of miR-1 in adult worms destroyed the morphological architecture of reproductive organs and reduced the subsequent oviposition, which may be due to the activation of apoptosis pathways. Through in silico analysis, 34 potential target genes of miR-1 were identified, including five ribosomal protein genes, called rp-s13, rp-l7ae, rp-l14, rp-l11 and rp-s24e. In vitro dual-luciferase reporter gene assays and miRNA overexpression experiments further validated that these ribosomal protein genes were directly regulated by miR-1. In contrast to the gene expression of miR-1, qRT-PCR and in situ hybridization experiments demonstrated these ribosomal protein genes were enriched in the sexual organs of adult females. Using RNA interference to silence the ribosomal protein genes in different developmental stages in a mouse model system, we demonstrated that these miR-1 target genes not only participated in the reproductive development of S. japonicum, but also were required for the growth and survival of the parasite in the early developmental stages. Taken together, our data suggested that miR-1 may affect the growth, reproduction and oviposition of S. japonicum by targeting the ribosomal protein genes, which provides insights for exploration of new anti-schistosome strategies.
成熟雌性血吸虫所产的卵是血吸虫病发病机制的主要成因，也是传播的关键因素。因此，阐明性成熟以及产卵机制可能会带来控制血吸虫病的新策略。微小RNA（miRNA）参与包括许多生物体繁殖在内的多种生物学过程，但其在血吸虫中的作用尚未得到充分表征。在此，我们研究了微小RNA - 1（miR - 1），它在日本血吸虫雌雄虫达到性成熟后均逐渐下调。定量逆转录PCR（qRT - PCR）结果显示，与体细胞组织相比，成年雌性血吸虫生殖器官中miR - 1的表达较低。在成年虫体中过表达miR - 1会破坏生殖器官的形态结构并减少随后的产卵量，这可能是由于凋亡途径的激活所致。通过计算机分析，鉴定出miR - 1的34个潜在靶基因，包括五个核糖体蛋白基因，分别称为rp - s13、rp - l7ae、rp - l14、rp - l11和rp - s24e。体外双荧光素酶报告基因测定和miRNA过表达实验进一步验证了这些核糖体蛋白基因受miR - 1直接调控。与miR - 1的基因表达相反，qRT - PCR和原位杂交实验表明这些核糖体蛋白基因在成年雌性的性器官中富集。在小鼠模型系统中利用RNA干扰沉默不同发育阶段的核糖体蛋白基因，我们证明这些miR - 1靶基因不仅参与日本血吸虫的生殖发育，而且在寄生虫早期发育阶段的生长和存活中也是必需的。综上所述，我们的数据表明miR - 1可能通过靶向核糖体蛋白基因影响日本血吸虫的生长、繁殖和产卵，这为探索新的抗血吸虫策略提供了思路。
