Delineating the Role of Host Cellular lncRNAs in Rotavirus Infection

Saha, Souhardya (2025) Delineating the Role of Host Cellular lncRNAs in Rotavirus Infection. Masters thesis, Indian Institute of Science Education and Research Kolkata.

Text (MS Dissertation of Souhardya Saha (20MS1470) 20MS147_Thesis_file.pdf - Submitted Version Restricted to Repository staff only Download (1MB)

Abstract

Long non-coding RNAs are regulatory RNAs of more than 200 nucleotides in length. They do not encode any proteins but are involved in regulation of gene expression and can modulate various cellular signaling pathways and processes. Research in disease biology including infectious diseases has revealed the dysregulation of several lncRNAs to modulate various cellular processes. To facilitate virus replication, pathogenesis and immune evasion, viruses hijack the host machinery to regulate the expression of host genes including the lncRNAs. Therefore, studying the host transcriptome in virus infection is an exciting and important area of research to understand the mechanism of virus pathogenesis. This study is aimed at understanding the regulation of host lncRNAs in rotavirus infection to determine the role of host lncRNAs in rotavirus pathogenesis. Based on a previously reported qPCR-based array of genes involved in inflammation and host immunity, and RNA sequencing, a set of lncRNAs were validated using qRT-PCR to be differentially expressed in rotavirus infection. Upon bioinformatic analysis using LncTarD 2.0 and literature studies, a significantly downregulated lncRNA MEG3 at early hours of infection was selected to study further. Overexpressing the MEG3 lncRNA significantly reduced the virus infection indicating its proviral role. Using the bioinformatics data, the interacting partners of MEG3 were identified and using RNA-immunoprecipitation, PI3K and STAT3 proteins were observed to interact with MEG3 lncRNA. The study was then focused on PI3K/Akt pathway as rotavirus was reported to modulate PI3K/Akt pathway to aid in virus pathogenesis. MEG3 overexpression resulted in lower activation of the PI3K/Akt axis confirmed by measuring protein levels through immunoblotting. Additionally, MEG3 overexpression promoted apoptosis indicated by the increasing levels of pro-apoptotic proteins Bax and cleaved caspase-3, and reduced level of anti-apoptotic Bcl2 protein. Therefore, it can be inferred that rotavirus downregulated MEG3 lncRNA to activate the pro-survival pathway of PI3K/Akt and inhibit apoptosis to allow virus replication inside host cells. This study uncovered the role of host lncRNAs, specifically MEG3 lncRNA in rotavirus infection which can be used to develop antiviral therapeutics against rotavirus in the future.

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