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Mpox virus infection and drug treatment modelled in human skin organoids

Nature Microbiology volume 8, pages 2067–2079 (2023)Cite this article

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Abstract

Mpox virus (MPXV) primarily infects human skin to cause lesions. Currently, robust models that recapitulate skin infection by MPXV are lacking. Here we demonstrate that human induced pluripotent stem cell-derived skin organoids are susceptible to MPXV infection and support infectious virus production. Keratinocytes, the predominant cell type of the skin epithelium, effectively support MPXV infection. Using transmission electron microscopy, we visualized the four stages of intracellular virus particle assembly: crescent formation, immature virions, mature virions and wrapped virions. Transcriptional analysis showed that MPXV infection rewires the host transcriptome and triggers abundant expression of viral transcripts. Early treatment with the antiviral drug tecovirimat effectively inhibits infectious virus production and prevents host transcriptome rewiring. Delayed treatment with tecovirimat also inhibits infectious MPXV particle production, albeit to a lesser extent. This study establishes human skin organoids as a robust experimental model for studying MPXV infection, mapping virus–host interactions and testing therapeutics.

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Fig. 1: MPXV infection in early-stage human skin organoids.
Fig. 2: MPXV infection in late-stage human skin organoids.
Fig. 3: MPXV infection in end-stage human skin organoids.
Fig. 4: Transcriptomic analysis of MPXV-infected organoids.
Fig. 5: MPXV–host interactions and response to tecovirimat treatment.
Fig. 6: MPXV infection and antiviral treatment in skin organoids cultured in ALI.

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Data availability

The data supporting the findings of this study are available within the paper, its supplementary information or its source data. RNA sequencing data are publicly available at https://doi.org/10.17026/dans-xj2-hhat. Mpox viral reads were mapped according to the reference genome (accession NC_063383.1). Source data are provided with this paper.

Code availability

The code used for transcriptomics analysis in this study is publicly available at https://doi.org/10.17026/dans-xzc-46u4

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Acknowledgements

We thank our colleagues at the hiPSC Hotel, Leiden University Medical Center, for providing hiPS cell lines. This work was supported by a VIDI grant (grant number 91719300) from the Dutch Research Council (NWO) to Q.P., and by the Novo Nordisk Foundation Center for Stem Cell Medicine supported by the Novo Nordisk Foundation, Denmark (grant number NNF21CC0073729) to K.R. K.R. is Chargé de Recherche at the Institut National de la Santé et de la Recherche Médicale (INSERM).

Author information

Author notes

  1. These authors contributed equally: Pengfei Li, Spyridon T. Pachis.

  2. These authors jointly supervised this work: Karine Raymond, Qiuwei Pan.

Authors and Affiliations

  1. Department of Gastroenterology and Hepatology, Erasmus MC-University Medical Center, Rotterdam, the Netherlands

    Pengfei Li, Guige Xu, Annemarie C. de Vries, Marco J. Bruno, Maikel P. Peppelenbosch & Qiuwei Pan

  2. Department of Anatomy and Embryology, Leiden University Medical Center, Leiden, the Netherlands

    Spyridon T. Pachis & Karine Raymond

  3. The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center, Leiden, the Netherlands

    Spyridon T. Pachis & Karine Raymond

  4. Department of Pathology, Erasmus MC-University Medical Center, Rotterdam, the Netherlands

    Rick Schraauwen

  5. Computational Bioscience Research Center, King Abdullah University of Science and Technology, Thuwal, Saudi Arabia

    Roberto Incitti & Intikhab Alam

  6. University of Grenoble Alpes, CEA, Inserm, IRIG, UA13 BGE, Biomics, Grenoble, France

    Karine Raymond

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Contributions

P.L., S.T.P., K.R. and Q.P. conceptualized the project. G.X., R.S., R.I. and I.A. developed the methodology. P.L., S.T.P., K.R., A.C.V. and Q.P. conducted the investigations. P.L., S.T.P., R.I., I.A. and K.R. conducted formal analysis. P.L., S.T.P., K.R. and Q.P. wrote the original draft. A.C.V., M.J.B. and M.P.P. reviewed and edited the manuscript. K.R. and Q.P. acquired funding.

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Correspondence to Karine Raymond or Qiuwei Pan.

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Extended data

Extended Data Fig. 1 Quantification of infectious viral titers in organoids at 1 hour and 7 days post-inoculation.

a, Infectious viral titers of organoids after 55 days of differentiation (n = 2 biological replicates). b, Infectious viral titers of organoids after 90 days of differentiation (n = 2 biological replicates).

Source data

Extended Data Fig. 2 Transmission electron microscopy analysis of skin organoids.

(a) and (b) show destroyed organelles, degrading cytoplasm (black arrowheads), and irregular chromatin condensation, alteration of nucleus membrane (red frame) in end-stage skin organoids. c, Representative skin characteristics was visualized in ALI-skin organoid, including skin dermis, collagen fibers, keratin bundles, desmosomes, and basement membrane.

Extended Data Fig. 3 Gene ontology analysis of skin organoids upon MPXV infection.

Top 30 significantly up- and down-regulated pathways by gene ontology analysis at day 7 post-inoculation, compared with the uninfected group. Three independent skin organoids were used in each group, P value determined by DESeq2.

Extended Data Fig. 4 Transcriptomic analysis of skin organoids upon 7 days infection, compared with uninfected or 1 h infection group.

a, Significantly up- and down-regulated genes by volcano plot analysis at day 7 post-inoculation, compared with the uninfected group. P value determined by DESeq2. b-e, Gene set enrichment analysis (GSEA) of KEGG pathways including autophagy_other (HSA04136), apoptosis (HSA04210), necroptosis (HSA04217), and ferroptosis (HSA04216) in skin organoids.

Extended Data Fig. 5 Total reads and mapped MPXV reads in different skin organoid groups by bulk-RNA sequencing.

Skin organoids were either not infected (control), infected for 1 hour (1 hour), infected for 7 days (7 day) or infected and treated with 5 μM of tecovirimat for 7 days (Tecovirimat), n = 3 per group. Data are presented as means of biological replicates ± SEM.

Source data

Extended Data Fig. 6 Low magnification images of the mpox virion (green) and integrin β4 subunit (red) immunostaining depicted in Fig. 6g.

White * indicates the potential autofluorescent signal of the cornified layer of the epithelium, which is distinguishable from the specific staining of mpox virion observed in the conditions where MPXV had been inoculated.

Supplementary information

Supplementary Information

Supplementary table and methods.

Reporting Summary

Source data

Source Data Fig. 1

qPCR and plaque assay source data.

Source Data Fig. 2

qPCR and plaque assay source data.

Source Data Fig. 3

qPCR and plaque assay source data.

Source Data Fig. 5

qPCR and statistical source data.

Source Data Fig. 6

qPCR and statistical source data.

Source Data Extended Data Fig. 1

Plaque assay source data.

Source Data Extended Data Fig. 5

RNA-sequencing mapped reads source data.

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Li, P., Pachis, S.T., Xu, G. et al. Mpox virus infection and drug treatment modelled in human skin organoids. Nat Microbiol 8, 2067–2079 (2023). https://doi.org/10.1038/s41564-023-01489-6

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