Expression Profiles of mRNA and lncRNA in HCT-8 Cells Infected With Cryptosporidium parvum IId Subtype.

Ting-Li Liu,Yan-Ling Yin,Ya-Jie Yuan,Xian-Chen Fan,Yun-Hui Li,Xue-Ting Wang,Guang-Hui Zhao,Long-Xian Zhang

doi:10.3389/fmicb.2018.01409

Abstract

Cryptosporidium parvum is one of the most important enteric protozoan pathogens, responsible for severe diarrhea in immunocompromised human and livestock. However, few effective agents were available for controlling this parasite. Accumulating evidences suggest that long non-coding RNA (lncRNA) played key roles in many diseases through regulating the gene expression. Here, the expression profiles of lncRNAs and mRNAs were analyzed in HCT-8 cells infected with C. parvum IId subtype using microarray assay. A total of 821 lncRNAs and 1,349 mRNAs were differentially expressed in infected cells at 24 h post infection (pi). Of them, all five types of lncRNAs were identified, including 22 sense, 280 antisense, 312 intergenic, 44 divergent, 33 intronic lncRNAs, and 130 lncRNAs that were not found the relationship with mRNAs’ location. Additionally, real-time polymerase chain reactions of 10 lncRNAs and 10 mRNAs randomly selected were successfully confirmed the microarray results. The co-expression and target prediction analysis indicated that 27 mRNAs were cis-regulated by 29 lncRNAs and 109 were trans-regulated by 114 lncRNAs. These predicted targets were enriched in several pathways involved in the interaction between host and C. parvum, e.g., hedgehog signaling pathway, Wnt signaling pathway, and tight junction, suggesting that these differentially expressed lncRNAs would play important regulating roles during the infection of C. parvum IId subtype.

Highlights

Cryptosporidium, one of the most important enteric parasites to cause diarrhea in human and animals (Holland, 1990; Nguyen et al, 2013; Koinari et al, 2014; Delafosse et al, 2015; Deshpande et al, 2015; Nakamura and Meireles, 2015), has been recognized as the leading cause of chronic diarrhea in HIV patients and was the second contributor of moderate-to-severe diarrhea in children during the first 2 years of life (Checkley et al, 2015)
To deeply understand the interaction between C. parvum and host, we systematically investigated the expression profiles of mRNA and long non-coding RNA (lncRNA) in human cells infected with the C. parvum IIdA19G1 subtype
To explore the impact of Chinese prevalent C. parvum subtype (IIdA19G1 subtype) on human cells, HCT-8 cells were exposed to C. parvum IIdA19G1 subtype for 24 h and collected for LncRNA+mRNA Human Gene Expression Microarray analysis

Summary

Introduction

Cryptosporidium, one of the most important enteric parasites to cause diarrhea in human and animals (Holland, 1990; Nguyen et al, 2013; Koinari et al, 2014; Delafosse et al, 2015; Deshpande et al, 2015; Nakamura and Meireles, 2015), has been recognized as the leading cause of chronic diarrhea in HIV patients and was the second contributor of moderate-to-severe diarrhea in children during the first 2 years of life (Checkley et al, 2015). In China, C. parvum has been detected in humans and animal species from provinces and one region (Qinling Mountain; Huang et al, 2014; Qi et al, 2015; Zhang X.X. et al, 2015; Cai et al, 2017). Of them, both subtypes IIa and IId were identified in China (Mi et al, 2013; Cui et al, 2014; Zhao et al, 2015). The total assembly length of C. parvum IIaA15G2R1 was longer than IIdA19G1, and the gene gains/losses (e.g., SKSRa, MEDLE family of secreted proteins, Insulinase-like proteases) and single nucleotide variants (SNVs) were found between the genomes of two subtypes (Widmer et al, 2012; Feng et al, 2017)

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Discussion

Conclusion