Integrative Transcriptomics and Proteomics Analyses to Reveal the Developmental Regulation of Metorchis orientalis: A Neglected Trematode With Potential Carcinogenic Implications.

Jun-Feng Gao,Qing-Bo Lv,Chun-Ren Wang,Qiao-Cheng Chang,Ying-Yu Chen,Rui-Feng Mao,Yang-Yuan Qiu,Yun-Yi Sun

doi:10.3389/fcimb.2021.783662

Abstract

Metorchis orientalis is a neglected zoonotic parasite of the gallbladder and bile duct of poultry, mammals, and humans. It has been widely reported in Asian, including China, Japanese, and Korea, where it is a potential threat to public health. Despite its significance as an animal and human pathogen, there are few published transcriptomic and proteomics data available. Transcriptome Illumina RNA sequencing and label-free protein quantification were performed to compare the gene and protein expression of adult and metacercariae-stage M. orientalis, resulting in 100,234 unigenes and 3,530 proteins. Of these, 13,823 differentially expressed genes and 1,445 differentially expressed proteins were identified in adult versus metacercariae. In total, 570 genes were differentially expressed consistent with the mRNA and protein level in the adult versus metacercariae stage. Differential gene transcription analyses revealed 34,228 genes to be expressed in both stages, whereas 66,006 genes showed stage-specific expression. Compared with adults, the metacercariae stage was highly transcriptional. GO and KEGG analyses based on transcriptome and proteome revealed numerous up-regulated genes in adult M. orientalis related to microtubule-based processes, microtubule motor activity, and nucleocytoplasmic transport. The up-regulated genes in metacercariae M. orientalis were mainly related to transmembrane receptor protein serine/threonine kinase activity, transmembrane receptor protein serine/threonine kinase signaling pathway. Transcriptome and proteome comparative analyses showed numerous up-regulated genes in adult stage were mainly enriched in actin filament capping, spectrin, and glucose metabolic process, while up-regulated genes in metacercariae stage were mainly related to cilium assembly, cilium movement, and motile cilium. These results highlight changes in protein and gene functions during the development of metacercariae into adults, and provided evidence for the mechanisms involved in morphological and metabolic changes at both the protein and gene levels. Interestingly, many genes had been proved associated with liver fibrosis and carcinogenic factors were identified highly expressed in adult M. orientalis, which suggests that M. orientalis is a neglected trematode with potential carcinogenic implications. These data provide attractive targets for the development of therapeutic or diagnostic interventions for controlling M. orientalis.

Highlights

The Opisthorchiidae is a large family of trematodes causing diseases with significant socioeconomic impacts in humans and animals in Asia and Europe, with more than 10 million people affected and ~680 million people estimated to be at risk of infection (Saijuntha et al, 2021)
The results showed that Differentially expressed genes (DEGs) of phosphoglycerate mutase, translation initiation factor 3 subunit B, and cathepsin F precursor were up-regulated in the adult stage versus metacercariae stage, whereas cytoplasmic 1, serpin, glutamine synthetase, elongation factor 1-gamma, and lactate dehydrogenase were up-regulated in metacercariae stage versus adult M. orientalis, which showed a similar expression trend to the transcriptome analysis, providing evidence on the reliability of the transcriptome sequencing results
Some down-regulated genes were enriched in gap junctions, glucagon signaling pathway, aldosterone synthesis and secretion, insulin secretion, and other pathways (Figure 5D). These results suggest that gene and protein expression levels were not always consistent, there was a high degree of consistency between the functions of DEGs and differentially expressed proteins (DEPs) in the different stages of M. orientalis

Summary

Introduction

The Opisthorchiidae is a large family of trematodes causing diseases with significant socioeconomic impacts in humans and animals in Asia and Europe, with more than 10 million people affected and ~680 million people estimated to be at risk of infection (Saijuntha et al, 2021). Opisthorchiidae flukes inhabit the biliary tract of the host, causing chronic diseases, including cholangitis, cholecystitis, cholelithiasis, and cholangiocarcinoma. Despite their significance, many of them have been neglected in terms of research and their control. Metorchis orientalis is a freshwater fluke and one such neglected member of the Opisthorchiidae. It mainly inhabits the gallbladder and bile duct of poultry and mammals, including humans. M. orientalis is endemic predominantly in regions of Korea and China (Qiu et al, 2017; Zhan et al, 2017; Sohn et al, 2021), where it has a wide geographical distribution across 19 provinces in China, representing a significant socioeconomic burden (Gao et al, 2018)

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Conclusion