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The Role of Differential Host Glycan Interactions in Rotavirus Cell Entry and Replication

Raque, Molly
http://orcid.org/0000-0003-2410-4798
http://rave.ohiolink.edu/etdc/view?acc_num=osu1669884288662793

Abstract Details

2022, Master of Science, Ohio State University, Veterinary Preventive Medicine.
Rotavirus A (RVA) is the primary cause of acute viral gastroenteritis in children and young animals globally; however, its replication and pathogenesis remain poorly understood. We have previously demonstrated contrasting modes of interactions with the host cell glycans for two prevalent porcine RVA strains: OSU G5P[7] (historically associated with severe disease in piglets) and G9P[13] (globally emerging variant in humans and swine). Specifically, OSU G5P[7] and G9P[13] strain replication was significantly decreased and significantly increased, respectively, following removal of terminal sialic acids (SA) by neuraminidase (NA) treatment. The latter contrasting effects coincided with the presence of distinct mutations found in the VP4 fusion region of these strains. In our first study, to clarify cellular mechanisms associated with these differential mechanisms of cell attachment/entry we conducted transcriptome analysis of porcine small intestinal enteroids (PIEs) infected with the two RVA strains with and without NA treatment. NA treatment of porcine intestinal enteroids alone, before individual RVA G9P[13]/OSU G5P[7] infection resulted in altered expression of genes associated with biological regulation, transporter activity, protein binding, and multicellular organismal processes. This was shown with significant contradicting impacts, with G9P[13] being significantly enhanced, and OSU G5P[7] replication being significantly inhibited. Cholesterol (a key component of the host plasma membranes) has been shown to play a critical role in RVA replication. To further improve our understanding of RVA pathogenesis taking into consideration RVA genotype-specific features, in our second study, we comparatively evaluated the effects of cholesterol and cholesterol-related additives [Methyl-β-cyclodextrin (MβCD), and diethylaminoethyl (DEAE), and bile acids (BAs)] on G9P[13] vs. OSU G5P[7] replication in vitro. Consistent with our previous findings, treatment with cholesterol and DEAE has increased replication of both strains. Further, our data demonstrated that depletion of cellular cholesterol levels by MβCD treatment resulted in decreased replication of RVA G9P[13] and OSU G5P[7]. Finally, in contrast to previous findings, we found that treatment of MA104 cells with bile acids led to enhanced replication of RVA strains G9P[13] and OSU G5P[7]. To further clarify the mechanisms behind the contrasting modes of RVA interaction with terminal SAs, in our third study, we have established a reverse genetics system (RGS) carrying 11 RVA OSU genes and demonstrated that replacement of VP4 in NA-sensitive OSU G5P[7] with the one from RVA G9P[13] led to the generation of a viable recombinant progeny virus. Thus, our study has expanded our understanding of the mechanisms of RVA cell attachment and entry and suggested that the differential entry mechanisms utilized by OSU G5P[7] and G9P[13] strains could have altered mechanisms of the cholesterol-dependent intracellular replication of these strains. Additionally, our study generated a robust RGS platform to study RVA pathogenesis and gene function and confirmed the pivotal role of the VP4 in genotype-specific interactions with the host-cell glycans.
Anastasia Vlasova (Advisor)
Linda Saif (Committee Member)
Qiuhong Wang (Committee Member)
259 p.
Biology; Genetics; Virology
Rotavirus A; Porcine Ileal Enteroids; Transcriptome Analysis; Pathogenesis; Reverse Genetics System; Sialic Acids

Recommended Citations

Citations

  • Raque, M. (2022). The Role of Differential Host Glycan Interactions in Rotavirus Cell Entry and Replication [Master's thesis, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1669884288662793

    APA Style (7th edition)

  • Raque, Molly. The Role of Differential Host Glycan Interactions in Rotavirus Cell Entry and Replication. 2022. Ohio State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1669884288662793.

    MLA Style (8th edition)

  • Raque, Molly. "The Role of Differential Host Glycan Interactions in Rotavirus Cell Entry and Replication." Master's thesis, Ohio State University, 2022. http://rave.ohiolink.edu/etdc/view?acc_num=osu1669884288662793

    Chicago Manual of Style (17th edition)

osu1669884288662793
159
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