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August 6, 2026VirusesOpen Access

EV-A71 and EV-D68 infect neuromuscular organoids, driving viral replication, gene downregulation, and caspase-3 cleavage.

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Why the study?

Molecular mechanisms underlying the neurovirulence and neuromuscular effects of Enterovirus A71 and D68 in severe neurological complications remain poorly understood.

Population

Human induced pluripotent stem cell-derived neuromuscular organoids

Comparison

EV-A71 infection vs EV-D68 infection

Design

In vitro experimental study

Key result

Infection of human neuromuscular organoids with EV-A71 and EV-D68 resulted in downregulation of neuronal and muscular gene networks, SNAP25 cleavage, and increased cleaved caspase-3 levels.

Authors

ASAmber J. SchottingIGInés García-RodríguezEFEline Freeze

Discussion

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Key expert perspectives

Captured external expert commentary on this paper, strongest first. Original sources are linked where available.

AAnonymousReviewer

“This manuscript investigates the susceptibility of neuromuscular organoids (NMOs) derived from induced pluripotent stem cells to infection by enterovirus A71 (EV-A71) and enterovirus D68 (EV-D68). The authors conclude that both viruses infect neuronal populations within NMOs. However, the data presented do not convincingly support this claim.”

MDPIAnonymous
DNDr. Nora M. ChapmanReviewer

“The authors have generated neuromuscular organoids (NMOs) from human stem cells as a model for human infection of enteroviruses with known neurologic pathology using a strain of enterovirus A71 (EV-A71) from a genotype associated with neurologic disease and the prototype enterovirus D68 (EV-D68) which was not associated with neurologic disease. Nevertheless, both viruses did infect the NMOs producing changes in transcription of the host cells and detectable production of viral RNA and virus. Earlier work by Hixon et al (Reference 22) had demonstrated that the Fermon strain of EV-D68 would replicate in human motor neurons, demonstrating that there was a likelihood of infection of the NMOs with this strain despite the differences of the prototype from the contemporary EV-D68 strains associated with outbreaks of acute flaccid myelitis (AFM).”

MDPIDr. Nora M. Chapman
DVDr. Venkatraman SiddharthanReviewer

“Schotting et al. clearly demonstrated the impact of non-polio enteroviruses on neuromuscular impairment using neuromuscular organoids.”

MDPIDr. Venkatraman Siddharthan

Overview

NMOs model EV neurovirulence in human tissue; leaves open clinical translation for AFM.

Key Points

  • This study aims to understand the molecular mechanisms of neuromuscular impairment caused by enteroviruses A71 and D68.
  • Utilized human induced pluripotent stem cell-derived neuromuscular organoids for cellular analysis
  • Conducted transcriptomic analysis to assess gene expression changes after viral infection
  • Evaluated protein-level changes related to synaptic function and apoptotic pathways
  • EV-A71 showed higher viral replication in NMOs compared to EV-D68.
  • Both viruses downregulated neuronal and associated muscle gene networks, with distinct impacts: EV-A71 more heavily suppressed neuronal pathways.
  • Increased levels of cleaved caspase-3 were observed, indicating activation of apoptotic signaling.

Structured PICO

P
Population
Human induced pluripotent stem cell-derived neuromuscular organoids (NMOs)
E
Exposure
Infection with Enterovirus A71 (EV-A71) or Enterovirus D68 (EV-D68)
O
Outcome
Cellular tropism, viral replication, transcriptomic changes, and pathogenic effects (cleavage of SNAP25, increased cleaved caspase-3)surrogate

Human neuromuscular organoids serve as a robust platform to model enterovirus-induced acute flaccid myelitis, revealing distinct virus-specific transcriptomic and protein-level pathogenic effects.

Cite This Study

Schotting et al. (2026) studied Enterovirus A71 and D68 infection. Enterovirus A71 (EV-A71) and enterovirus D68 (EV-D68) infection was evaluated on Cellular tropism and pathogenic effects. Infection of human neuromuscular organoids with EV-A71 and EV-D68 resulted in downregulation of neuronal and muscular gene networks, SNAP25 cleavage, and increased cleaved caspase-3 levels.

synapsesocial.com/papers/6a74379d764cddc9499d508chttps://doi.org/10.3390/v18080853
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