Dataset Information

Impact of hepatitis E virus host- and virus-derived insertions on the antiviral response in primary human hepatocytes

ABSTRACT: Background & Aims Despite the global burden of hepatitis E virus (HEV, Paslahepevirus balayani) infection, host-virus interactions underlying HEV pathogenesis and progression to chronicity remain incompletely understood. This study aimed to characterize the host transcriptomic response to infection with HEV variants harboring different host- and virus-derived insertions in the hypervariable region (HVR), with a focus on innate immune activation. As model system with high physiological relevance, we used primary human hepatocytes (PHHs). Methods We performed bulk RNA sequencing on PHHs from three donors infected with HEV-3 variants harboring different HVR insertions (RPS17, HVR duplication, SERPINA1, TRIM22), which have been previously identified in vivo in a chronic HEV patient. To compare the replication fitness of the different insertion variants, we quantified progeny virus titers and viral RNA reads in the cellular transcriptome. Host transcriptomic responses were analyzed in detail at multiple time points post infection to assess virus-induced differential gene expression. To further characterize the response to HEV infection with different HVR insertion variants, we performed gene ontology term enrichment analysis and identified individual HEV-induced genes. Results Progeny virus titers at 72 hours post infection (h p.i.) were comparable across all PHH donors and HVR insertion variants. Donors 1 and 2 mounted robust host responses to HEV infection, characterized by widespread gene deregulation and interferon stimulated gene (ISG) upregulation at 72 h p.i. In contrast, donor 3 exhibited minimal virus-induced transcriptional changes. Across all variants, the HVR duplication insertion variant consistently triggered a weaker host response compared to RPS17, SERPINA1 and TRIM22 insertion variants, with significantly fewer differentially expressed genes and reduced ISG induction. Gene ontology term analysis revealed enrichment of antiviral pathways upon infection with the different insertion variants, which was less pronounced for the HVR duplication variant. Conclusions These findings reveal that HEV HVR rearrangement variants elicit distinct host response patterns, with the HVR duplication variant showing notably diminished innate immune activation compared to host-derived insertions. Importantly, our results suggest a positive correlation of viral replication and host response induction. This indicates that the HVR may play a crucial role in shaping host-virus interactions and emphasizes the importance of comprehensive HVR characterization in understanding HEV pathogenesis. Impact and implications This study provides novel insights into HEV-host interactions, characterizing donor-specific and variant-dependent differences in antiviral responses. Our findings highlight the importance of understanding the role of the HVR in chronic [URG3.1][SS3.2]HEV infection, a major clinical challenge particularly in immunocompromised patients where persistent viremia can lead to progressive liver damage. The observed variations in innate immune activation patterns across different donors and viral variants suggest a critical role of innate immunity in shaping HEV pathogenesis and potentially determining infection outcomes.

ORGANISM(S): Homo sapiens

PROVIDER: GSE347085 | GEO | 2026/09/18

REPOSITORIES: GEO

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Project description:Background & Aims: Other than hepatitis B or C virus infection, Hepatitis E virus (HEV) infection is generally asymptomatic or leads to acute and self-limiting hepatitis. However, the mechanism of host cell defense against HEV is unclear. Viruses are known to perturb host cellular metabolism to enable their replication and spread. AMP-activated protein kinase (AMPK) activation is crucial for the regulation of cell homeostasis. We thus investigated the role of AMPK in HEV infection. Methods: Huh7, THP1 and HepG2 cells inoculated with infectious HEV viral particle or Huh7 and organoids transfected with in vitro generated subgenomic or full-length GT3 (Kernow-C1 p6 strain) HEV RNA, namely, p6Luc or p6 were used to model HEV infection. Viral replication and genes expression were quantified. Activation of AMPK, innate immune response and autophagy process were assessed. Results: We found HEV infection can trigger AMPK activation by phosphorylation of AMPK at threonine 172 by transfecting HEV viral RNA into host cells or inoculating host cells with infectious HEV viral particle. The activation of AMPK is associated with HEV induced mitochondrial damage and ATP deficiency. Pharmacological activation of AMPK using 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) attenuated HEV replication, which was reversed by an AMPK inhibitor (compound C). Lentivirus-mediated knockdown of AMPK provided further evidence that AMPK has an antiviral effect on HEV replication. These results suggested that AMPK activation is a potent strategy of host cells for HEV clearance. Consistent with its antiviral effect, AMPK activation potentiated the expression of genes with antiviral properties (e.g., IFNs, ISG15, and IRF9) and inhibited inflammatory response (e.g., NF-KB NLRP3 and IL-1β). Meanwhile, HEV and activated AMPK also decreased autophagosome accumulation by decreasing induction of autophagy and autophagic degradation. Consistently, we found inhibition of AMPK efficiently augmented HEV induced autophagosome accumulation, evidenced by a marked increase in LC3II. Our previous study showed that rapamycin, an activator of autophagic induction by inhibiting mTOR, and Bafilomycin A1, an inhibitor of autophagic degradation, has a potent pro-HEV effect under AICAR treatment. Moreover, Wortmannin inhibiting autophagic induction recover AMPK inhibitor induced HEV replication. Together, these results suggested that HEV induced AMPK activation can serve to protect HEV infected cells from HEV infection by attenuating autophagosome and promoting innate immunity. Conclusions: Here we show that HEV infection can activate AMPK phosphorylation, which attenuates autophagosome accumulation and increases innate immune signaling. Thus, the AMPK activation in response to HEV infection is critical in host cells for rapid viral clearance by coordinating autophagic process and establishing persistent antiviral immunity.

Dataset Information

Impact of hepatitis E virus host- and virus-derived insertions on the antiviral response in primary human hepatocytes

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets