Abstract
Poliomyelitis is caused by poliovirus (PV), a positive strand non-enveloped virus. Since its discovery in the 1950s, several cell culture and molecular methods have been developed to detect and characterize the various strains of PV. Here, we provide an accurate and standardized protocol to differentiate human embryonic stem cells (hESCs) toward engineered neural tissue enriched with motor neurons (MN ENTs). These MN ENTs expressed markers of motor neuron CHAT and Hb-9 as revealed by immunofluorescence staining and quantitative RT-PCR. Interestingly, our results suggest that motor neurons are responsible for the permissiveness of poliovirus within the MN ENTs. Moreover, our study revealed the molecular events occurring upon PV-3 infection in the MN ENTs and highlighted the modulation of a set of genes involved in EGR-EP300 complex. Collectively, we report the development of a reliable in vitro model to investigate the pathophysiology of PV infection, allowing to both design and assess novel therapeutic approaches against PV infection.
Highlights
Poliovirus (PV) is a small non-enveloped virus with a single-strand positive genomic RNA classified in the Enterovirus C species of the Picornaviridae family (Couderc et al, 1989)
Given the known tropism of poliovirus for MNs, we developed a protocol to enrich engineered neural tissues with motor neurons (MN ENTs), as outlined in Supplementary Figure 1A
Human lymphocytes B were used as negative controls. (C) Immunofluorescence showed neuronal nucleispecific protein (NeuN), CHAT, and HB-9-immunoreactive cells present in the whole MN engineered neural tissue (MN ENT)
Summary
Poliovirus (PV) is a small non-enveloped virus with a single-strand positive genomic RNA classified in the Enterovirus C species of the Picornaviridae family (Couderc et al, 1989). Due to its fecal-oral transmission route, PV infection is most often present in high density population with subpar sanitation systems (Falconer and Bollenbach, 2000). PV infects the gray matter of the anterior horn of the spinal cord, from which its name derived (Greek polios = gray, myelos = matter). In 1–2% of infected individuals, the virus enters the central nervous system (CNS) and replicates in motor neurons (MN) within the spinal cord, brain stem, or motor cortex (Kew et al, 2005; Racaniello, 2006). Survivors of poliomyelitis often remain disabled (Mueller et al, 2005). Since the 1950’s, major vaccination campaigns allowed to decrease the incidence of poliomyelitis (Cochi et al, 2016). Despite few reported cases of poliomyelitis, several reports acknowledge the possible eradication of PV in the decade
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