A human-derived 3D brain organoid model to study JC virus infection

Abstract

Progressive multifocal leukoencephalopathy (PML) is a frequent neurological complication in immunosuppressed patients. PML is caused by the JC virus (JCV), a neurotropic DNA polyomavirus that infects oligodendrocytes and astrocytes, causing inflammation and demyelination which lead to neurological dysfunction. The pathogenesis of PML is poorly understood due to the lack of in vitro or animal models to study mechanisms of disease as the virus most efficiently infects only human cells. We developed a human-derived brain organotypic system (also called brain organoid) to model JCV infection. The model was developed by using human-induced pluripotent stem cells (iPSC) and culturing them in 3D to generate an organotypic model containing neurons, astrocytes, and oligodendrocytes which recapitulates aspects of the environment of the human brain. We infected the brain organoids with the JCV MAD4 strain or cerebrospinal fluid of a patient with PML. The organoids were assessed for evidence of infection by qPCR, immunofluorescence, and electron microscopy at 1, 2, and 3 weeks post-exposure. JCV infection in both JCV MAD4 strain and PML CSF-exposed brain organoids was confirmed by immunocytochemical studies demonstrating viral antigens and electron microscopy showing virion particles in the nuclear compartment of oligodendrocytes and astrocytes. No evidence of neuronal infection was visualized. Infection was also demonstrated by JCV qPCR in the virus-exposed organoids and their media. In conclusion, the brain organoid model of JCV infection establishes a human model suitable for studying the mechanisms of JCV infection and pathogenesis of PML and may facilitate the exploration of therapeutic approaches.

Human Brain Organoids as Models for Central Nervous System Viral Infection

“…The immune response of PD cerebral organoids was altered compared to the non-PD organoids, highlighting the need for studies on the influence of host genetic variation on host-pathogen interactions [ 95 ]. John Cunningham virus (JCV), a DNA polyomavirus, was studied once in cerebral organoids [ 96 ]. JCV causes opportunistic infections in immunocompromised individuals, leading to progressive multifocal leukoencephalitis, a fatal demyelinating disease [ 97 , 98 ].…”

“…JCV causes opportunistic infections in immunocompromised individuals, leading to progressive multifocal leukoencephalitis, a fatal demyelinating disease [ 97 , 98 ]. Brain organoids have been shown to be a suitable model for JCV pathogenesis with JCV infection of brain organoids, resulting in viral replication and infection of astrocytes and oligodendrocytes, while neurons were not infected [ 96 ].…”

The Inhibition of DNA Viruses by the Amphibian Antimicrobial Peptide Temporin G: A Virological Study Addressing HSV-1 and JPCyV

“…However, in vivo models for cerebral JCPyV-related diseases are currently not available. Nevertheless, a human-derived brain organotypic system has been recently developed to model virus infection and virus-induced brain pathologies [45,46].…”

iPSC-derived three-dimensional brain organoid models and neurotropic viral infections

“…However, JCV infection in this model resulted in virus replication mainly in human astrocytes with signs of demyelination not imputable to viral lytic infection due to the limited human oligodendrocyte incorporation. Recently, Barreras and colleagues (Barreras et al 2022 ) have established a promising human brain organoid model to study JCV infection. The brain organoids consisted of neurons (70%), astrocytes (20%) and oligodendrocytes (10%) with a diameter of 300–350 μm.…”