632 - Contemporary Enterovirus-D68 Infects Human Spinal Cord Organoids

Poster Number: 632
Publication Number: 632.226

Megan C. Freeman, UPMC Childrens Hospital of Pittsburgh, Pittsburgh, PA, United States; Gabrielle Aguglia, UPMC Childrens Hospital of Pittsburgh, Pittsburgh, PA, United States; Carolyn Coyne, Duke University School of Medicine, Durham, NC, United States; Terence Dermody, UPMC Childrens Hospital of Pittsburgh, Pittsburgh, PA, United States; John V.. Williams, Children's Hospital of Pittsburgh/University of Pittsburgh, Pittsburgh, PA, United States

Background: Enterovirus D68 (EV-D68) is a nonpolio enterovirus associated with severe respiratory illness and acute flaccid myelitis (AFM), a polio-like illness causing paralysis in children. The emergence of AFM outbreaks has been associated with increased circulation and genetic diversity of EV-D68 since 2014, although the virus was discovered in the 1960s. The mechanisms by which EV-D68 targets the central nervous system are unknown. Enteroviruses are human pathogens, without routine infection of other animal species, making a human model of the central nervous system essential for understanding pathogenesis.

Objective: We sought to develop two new human spinal cord organoid (hSCO) models for EV-D68 infection derived from induced, pluripotent stem cell (iPSC) lines.

Design/Methods: We built upon previous protocols for creating spinal cord organoids from induced, pluripotent stem cells and evaluated their capacity for infection with historic and contemporary strains of EV-D68. One model is comprised primarily of spinal motor neurons, while the other includes spinal motor neurons, interneurons, and glial cells. We evaluated morphology, cell marker expression, replication kinetics, and cellular death in this model during infection.

Results:

hSCO can be productively infected with contemporary, but not historic, strains of EV-D68 and produce extracellular virus for at least two weeks without appreciable cytopathic effect. By comparison, infection with hSCO with echovirus 11, a common cause of neonatal meningoencephalitis, causes significant structural destruction and apoptosis.

Conclusion(s): We have established a human spinal cord organoid model for infection with EV-D68. Our results suggest that EV-D68 infection is not the sole driver of neuronal death in the spinal cord in AFM and that secondary injury from the immune response likely contributes to pathogenesis.This new hSCO model for EV-D68 infection will contribute to understanding how EV-D68 mediates damage to the human spinal cord, which could lead to new therapeutic and prophylactic strategies for this virus.