bioRxiv Science⌕ Search
Home/Archive/bioRxiv

bioRxiv · 10.1101/2025.09.19.677307

Hepatitis E ORF2 blocks trophoblast autophagy to induce miscarriage via LC3B binding rather than PI3K/Akt/mTOR pathway suppression

Chen, Y.·Yang, Y.·Bai, Q.·Tian, X.·Zhou, C.·Lu, X.·Liu, T.
View original source ↗Export metadata ↓

Abstract

Hepatitis E virus (HEV) is a zoonotic pathogen that can infect pregnant women and cause adverse pregnancy outcomes, including miscarriage and preterm delivery. The previous study demonstrated that HEV genotype 3 (HEV-3) inhibits complete autophagic flux in both mouse placental tissue and human trophoblast cells (JEG-3), evidenced by reduced expression of ATG proteins (including LC3, Beclin1, ATG4B, ATG5, and ATG9A) and accumulation of p62. However, the specific regulatory pathway involved remains unclear. Thus, eukaryotic expression vectors for HEV open reading frames (ORFs) were constructed, and ORF2 and ORF3 proteins were transiently overexpressed in JEG-3 cells via liposome transfection.. While both ORF2 and ORF3 significantly reduced LC3B protein levels (p < 0.01), only ORF2 induced p62 accumulation (p < 0.01), indicative of autophagic inhibition, which indicates that ORF2 was the key viral protein mediating autophagy suppression in JEG-3. The results of WB and RT-qPCR show that ORF2 suppressed the PI3K/Akt/mTOR pathway while enhancing nuclear translocation of TFEB (p < 0.01) and AMPK phosphorylation (p < 0.01), suggesting paradoxical activation of upstream autophagy regulators. Through co-transfection of mCherry-LC3 with ORF2, co-localization studies, and AlphaFold 3-based intermolecular interaction predictions, we proposed that ORF2 directly binds LC3B to block autophagosome formation. Finally, co-immunoprecipitation confirmed physical interaction between HEV ORF2 and LC3B, elucidating the molecular mechanism of HEV-induced autophagy suppression in trophoblasts. These findings reveal the molecular mechanism by which HEV inhibits autophagy leading to miscarriage in mice, providing new insights into HEV-induced reproductive damage.

Source connections

Explore related subjects

pathologyPreprint

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Chen, Y., Yang, Y., Bai, Q., Tian, X., Zhou, C., Lu, X., Liu, T.. 2025-09-21. Hepatitis E ORF2 blocks trophoblast autophagy to induce miscarriage via LC3B binding rather than PI3K/Akt/mTOR pathway suppression. https://doi.org/10.1101/2025.09.19.677307

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

bioRxiv2026

Prevention of Unc13a cryptic splicing is sufficient to preserve memory

TDP-43 dysfunction is thought to underlie frontotemporal dementia and limbic-predominant age-related TDP-43 encephalopathy, neurodegenerative dementias currently without effective therapy. Therapeutic strategies are designed to correct individual cryptic targets of TDP-43, such as UNC13A, whereby its cryptic splicing compromises synaptic function, yet the sufficiency of such an approach to prevent memory deficits is unclear. Using a forebrain neuron-specific TDP-43 knockout mouse model that recapitulates TDP-43 dysfunction occurring during early stages of human disorders, we found here that prevention of cryptic splicing to include that of Unc13a attenuated memory deficits. We show that genetic ablation of Unc13a cryptic exon solely in such TDP-43 knockout mice is sufficient to preserve cognition, supporting the clinical value of targeting UNC13A to mitigate memory deficits. Prevention of cryptic splicing of multiple targets of TDP-43 additionally attenuate neuron loss. For optimal outcomes in TDP-43 related dementias, these findings thus strongly support strategies designed to repress cryptic splicing of multiple targets of TDP-43, including UNC13A.

pathology↗
bioRxiv2026

Social-Cognitive Dysregulation Model of Misophonia: Perspective from a Behavioural Study

Misophonia is increasingly conceptualized as more than a disorder of sound tolerance, with trigger over-reactivity shaped by the social meaning of sounds, inferred intentions, and representations of others actions. We tested a social-cognitive dysregulation model of misophonia in (N = 341) adults using behavioural measures of Theory of Mind and emotion recognition, alongside measures of reflective functioning, empathy, alexithymia, and mimicry. Dimensional associations with misophonia severity and its five different dimensions were examined while accounting for age, sex, sound sensitivity, and anxiety/depressive symptoms. Increased misophonia severity was associated with less accurate and slower mental-state inference and emotion recognition. ToM accuracy effects were evident for more complex, cognitive, and affective mentalizing, but not for simpler mentalizing or physical control judgments, while emotion-recognition accuracy differences emerged for positive but not negative stimuli. Greater severity was also characterized by reduced certainty and greater uncertainty about mental states, greater difficulty identifying one s own feelings, and elevated alexithymia, whereas global self-reported empathy was largely preserved. Misophonia severity further predicted a greater propensity to mimic trigger-producing actions or sounds; 41% of participants exceeding the S-Five clinical cutoff (> 87) endorsed mimicry, which was particularly associated with a subjective restoration of control. Findings remained robust following influential-case sensitivity analyses. These results reveal a selective disturbance in self-other representation spanning mentalizing, emotion decoding, emotional self-representation, and embodied regulatory processes. They position misophonia within a broader social-cognitive framework in which auditory-affective reactivity may intersect with altered inferential and sensorimotor processing, while stopping short of causal inference.

pathology↗
bioRxiv2026

Lamin A/C depletion from myofibers and satellite cells in mice reveals selective muscle pathology

Mutations in the laminA/C gene (LMNA), which encodes the nuclear lamina proteins lamin A and lamin C (lamin A/C), have been linked to different human diseases affecting different tissues. Most LMNA mutations cause cardiomyopathy and muscular dystrophy, such as autosomal dominant Emery-Dreifuss muscular dystrophy. Recent studies to understand striated muscle laminopathies have taken advantage of Lmna conditional knockout mice to examine the effects of lamin A/C depletion in cardiomyocytes and cardiac fibroblasts. However, the role of lamin A/C in skeletal muscle has largely been uncharacterized using conditional knockout mice. We used different mouse lines to deplete lamin A/C from specific cell types in striated muscle. Lamin A/C depletion from fetal myofibers and cardiomyocytes led to no observable phenotype in the skeletal muscles despite leading to dramatic heart dilation and early lethality. Depletion of lamin A/C from both skeletal myofibers and satellite cells was lethal, with the most dramatic myopathic abnormalities observed in the intrinsic muscles of the tongue. The presence of lamin A/C in skeletal muscle satellite cells prevented the development of lethal myopathy when the proteins were deleted only from differentiated myofibers. Overall, our results provide a foundation for understanding the roles of lamin A/C in muscle maintenance and development, including the variable skeletal muscle involvement and much more invariant cardiomyopathy in patients with LMNA mutations.

pathology↗