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Updated: Oct 1, 2025
Modelling Zika Virus Infection of the Developing Human Brain In Vitro Using Stem Cell Derived Cerebral Organoids
Published on: September 19, 2017
A human-derived 3D brain organoid model to study JC virus infection
Paula Barreras1, David Pamies2, Maria Chiara Monaco3
1Department of Neurology, Division of Neuroimmunology, Johns Hopkins University, Baltimore, USA.
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.
Insights
Researchers developed a human brain organoid model to study Progressive Multifocal Leukoencephalopathy (PML), caused by the JC virus (JCV). This model successfully replicates JCV infection in human brain cells, offering new avenues for understanding PML pathogenesis and therapies.
Area of Science:
- Neuroscience
- Virology
- Stem Cell Biology
Background:
- Progressive Multifocal Leukoencephalopathy (PML) is a severe neurological disease affecting immunosuppressed individuals.
- PML is caused by the JC virus (JCV), which infects human oligodendrocytes and astrocytes, leading to demyelination and neurological deficits.
- Current understanding of PML pathogenesis is limited by the lack of suitable in vitro and animal models due to JCV's specific tropism for human cells.
Purpose of the Study:
- To develop a human-derived brain organotypic system (brain organoid) for modeling JC virus (JCV) infection.
- To investigate the mechanisms of JCV infection and the pathogenesis of Progressive Multifocal Leukoencephalopathy (PML) in a human cellular context.
Main Methods:
- Human-induced pluripotent stem cells (iPSC) were cultured in 3D to generate brain organoids containing neurons, astrocytes, and oligodendrocytes.
- Brain organoids were infected with the JCV MAD4 strain or cerebrospinal fluid from a PML patient.
- Infection was assessed using qPCR, immunofluorescence, and electron microscopy at 1, 2, and 3 weeks post-exposure.
Main Results:
- JC virus (JCV) infection was confirmed in brain organoids exposed to JCV MAD4 or PML patient cerebrospinal fluid.
- Immunocytochemistry and electron microscopy revealed viral antigens and virion particles within oligodendrocytes and astrocytes, but not neurons.
- Quantitative PCR (qPCR) detected JCV in the infected organoids and their surrounding media.
Conclusions:
- The developed brain organoid system serves as a viable human model for studying JCV infection.
- This model facilitates research into the pathogenesis of Progressive Multifocal Leukoencephalopathy (PML).
- The model holds potential for exploring and evaluating therapeutic strategies against PML.
