Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais-UFMG, Belo Horizonte, Brazil.
Instituto René Rachou, Fundação Oswaldo Cruz-FIOCRUZ, Belo Horizonte, Brazil.
Front Immunol. 2019 Jan 24;10:21. doi: 10.3389/fimmu.2019.00021. eCollection 2019.
Eukaryotic protein kinases (ePKs) are good medical targets for drug development in different biological systems. ePKs participate in many cellular processes, including the p38 MAPK regulation of homeostasis upon oxidative stress. We propose to assess the role of Smp38 MAPK signaling pathway in development and protection against oxidative stress, parasite survival, and also to elucidate which target genes have their expression regulated by Smp38 MAPK. After a significant reduction of up to 84% in the transcription level by Smp38 MAPK gene knockdown, no visible phenotypic changes were reported in schistosomula in culture. The development of adult worms was tested in mice infected with the Smp38 knocked-down schistosomula. It was observed that Smp38 MAPK has an essential role in the transformation and survival of the parasites as a low number of adult worms was recovered. Smp38 knockdown also resulted in decreased egg production, damaged adult worm tegument, and underdeveloped ovaries in females. Furthermore, only ~13% of the eggs produced developed into mature eggs. Our results suggest that inhibition of the Smp38 MAPK activity interfere in parasites protection against reactive oxygen species. Smp38 knockdown in adult worms resulted in 80% reduction in transcription levels on the 10th day, with consequent reduction of 94.4% in oviposition . In order to search for Smp38 MAPK pathway regulated genes, we used an RNASeq approach and identified 1,154 DEGs in Smp38 knockdown schistosomula. A substantial proportion of DEGs encode proteins with unknown function. The results indicate that Smp38 regulates essential signaling pathways for the establishment of parasite homeostasis, including genes related to antioxidant defense, structural composition of ribosomes, spliceosomes, cytoskeleton, as well as, purine and pyrimidine metabolism pathways. Our data show that the Smp38 MAPK signaling pathway is a critical route for parasite development and may present attractive therapeutic targets for the treatment and control of schistosomiasis.
真核蛋白激酶（ePKs）是不同生物系统药物开发的良好医学靶点。ePKs 参与许多细胞过程，包括 p38 MAPK 对氧化应激下内稳态的调节。我们建议评估 Smp38 MAPK 信号通路在发育和对氧化应激、寄生虫存活的保护中的作用，也阐明哪些靶基因的表达受 Smp38 MAPK 调节。在 Smp38 MAPK 基因敲低导致转录水平显著降低达 84%后，培养中的毛蚴没有报告明显的表型变化。在感染 Smp38 敲低毛蚴的小鼠中测试了成虫的发育。观察到 Smp38 MAPK 在寄生虫的转化和存活中起着至关重要的作用，因为回收的成虫数量很少。Smp38 敲低也导致产卵减少、成虫表皮受损和雌性卵巢发育不良。此外，只有约 13%的产生的卵发育成熟。我们的结果表明，抑制 Smp38 MAPK 活性会干扰寄生虫对活性氧的保护。成虫中的 Smp38 敲低导致第 10 天转录水平降低 80%，随后产卵减少 94.4%。为了寻找 Smp38 MAPK 通路调节的基因，我们使用了 RNAseq 方法，在 Smp38 敲低毛蚴中鉴定到 1154 个差异表达基因。大量差异表达基因编码功能未知的蛋白质。结果表明，Smp38 调节寄生虫内稳态建立的重要信号通路，包括与抗氧化防御、核糖体结构组成、剪接体、细胞骨架以及嘌呤和嘧啶代谢途径相关的基因。我们的数据表明，Smp38 MAPK 信号通路是寄生虫发育的关键途径，可能为治疗和控制血吸虫病提供有吸引力的治疗靶点。
