Xie Yuxiang, Wang Xiaoling, Cheng Shaoyun, Liu Wanling, Yi Cun, You Yanmin, Zhang Wei, Wang Yuepeng, Tang Enlu, Wang Jipeng, Hu Wei
State Key Laboratory of Genetic Engineering, Ministry of Education Key Laboratory of Contemporary Anthropology, School of Life Sciences, Fudan University, Shanghai, China.
Department of Infectious Diseases, Huashan Hospital, Fudan University, Shanghai, China.
PLoS Pathog. 2025 Mar 28;21(3):e1013014. doi: 10.1371/journal.ppat.1013014. eCollection 2025 Mar.
Schistosomiasis affects more than 250 million people worldwide and is one of the neglected tropical diseases. Currently, the treatment of schistosomiasis relies on a single drug-praziquantel-which has led to increasing pressure from drug resistance. Therefore, there is an urgent need to find new treatments. The development of genome sequencing has provided valuable information for understanding the biology of schistosomes. In the genome of Schistosoma japonicum, approximately 11% of the protein-coding sequences are uncharacterized genes (UGs) annotated as "hypothetical protein" or "protein of unknown function." These poorly understood genes have been unjustifiably neglected, although some may be essential for the survival of the parasites and serve as potential drug targets. In this study, we systematically mined the highly expressed UGs in both genders of this parasite throughout key developmental stages in their mammalian host, using our previously published S. japonicum genome and RNA-seq data. By employing in vitro RNA interference (RNAi), we screened 126 UGs that lack homologs in Homo sapiens and identified 8 that are essential for the parasite vitality. We further investigated two UGs, Sjc_0002003 and Sjc_0009272, which resulted in the most severe phenotypes. Fluorescence in situ hybridization demonstrated that both genes were expressed throughout the body without sex bias. Silencing either Sjc_0002003 or Sjc_0009272 reduced the cell proliferation in the body. Furthermore, in vivo RNAi indicated both genes are required for the growth and survival of the parasites in the mammalian host. For Sjc_0002003, we further characterize the underlying molecular cause of the observed phenotype. Through RNA-seq analysis and functional studies, we revealed that silencing Sjc_0002003 reduces the expression of a series of intestinal genes, including Sjc_0007312 (hypothetical protein), Sjc_0008276 (vha-17), Sjc_0002942 (PLA2G15), and Sjc_0003646 (SJCHGC09134 protein), leading to gut dilation. Our work highlights the importance of UGs in schistosomes as promising targets for drug development in the treatment of the schistosomiasis.
血吸虫病影响着全球超过2.5亿人，是被忽视的热带病之一。目前，血吸虫病的治疗依赖于单一药物——吡喹酮，这导致了来自耐药性的压力不断增加。因此，迫切需要寻找新的治疗方法。基因组测序的发展为理解血吸虫的生物学特性提供了有价值的信息。在日本血吸虫的基因组中，大约11%的蛋白质编码序列是未表征的基因（UGs），被注释为“假定蛋白”或“功能未知的蛋白”。这些了解甚少的基因一直被不合理地忽视，尽管其中一些可能对寄生虫的生存至关重要，并可作为潜在的药物靶点。在本研究中，我们利用我们之前发表的日本血吸虫基因组和RNA测序数据，系统地挖掘了这种寄生虫在其哺乳动物宿主体内关键发育阶段两性中高表达的UGs。通过体外RNA干扰（RNAi），我们筛选了126个在智人中缺乏同源物的UGs，并鉴定出8个对寄生虫活力至关重要的基因。我们进一步研究了两个UGs，Sjc_0002003和Sjc_0009272，它们导致了最严重的表型。荧光原位杂交表明这两个基因在全身表达且无性别偏向。沉默Sjc_0002003或Sjc_0009272都会降低体内的细胞增殖。此外，体内RNAi表明这两个基因是寄生虫在哺乳动物宿主体内生长和存活所必需的。对于Sjc_0002003，我们进一步表征了观察到的表型的潜在分子原因。通过RNA测序分析和功能研究，我们发现沉默Sjc_0002003会降低一系列肠道基因的表达，包括Sjc_0007312（假定蛋白）、Sjc_0008276（vha-17）、Sjc_0002942（PLA2G15）和Sjc_0003646（SJCHGC09134蛋白），从而导致肠道扩张。我们的工作突出了UGs在血吸虫中作为治疗血吸虫病药物开发的有前景靶点的重要性。
