Sofosbuvir protects human brain organoids against SARS-CoV-2

  1. Pinar Mesci
  2. Angela Macia
  3. Aurian Saleh
  4. Laura Martin-Sancho
  5. Xin Yin
  6. Cedric Snethlage
  7. Simoni Avansini
  8. Sumit K. Chanda
  9. Alysson Muotri

Reviewed by

Read the full article See related articles

Discuss this preprint

Start a discussion What are Sciety discussions?

Listed in

Log in to save this article

Abstract

COVID-19 was rapidly declared a pandemic by the World Health Organization, only three months after the initial outbreak in Wuhan, China. Early clinical care mainly focused on respiratory illnesses. However, a variety of neurological manifestations in both adults and newborns are also emerging. To determine whether SARS-CoV-2 could target the human brain, we infected iPSC-derived human brain organoids. Our findings show that SARS-CoV-2 was able to infect and kill neural cells, including cortical neurons. This phenotype was accompanied by impaired synaptogenesis. Finally, Sofosbuvir, an FDA-approved antiviral drug, was able to rescue these alterations. Given that there are currently no vaccine or antiviral treatments available, urgent therapies are needed. Our findings put Sofosbuvir forward as a potential treatment to alleviate COVID-19-related neurological symptoms.

One Sentence Summary

SARS-CoV-2 infection causes neuronal death and impaired synaptogenesis, both rescued by Sofosbuvir treatment.

Article activity feed

  1. ScreenIT

    SciScore for 10.1101/2020.05.30.125856: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    Institutional Review Board StatementConsent: Donated healthy fibroblasts were obtained via skin biopsies from patients after informed consent was appropriately given under protocols approved by the University of California, San Diego Institutional Review Board (#141223ZF).
    IRB: All experiments were approved and performed under the Institutional Review Boards (IRB) and Embryonic Stem Cell Research Oversight (ESCRO) guidelines and regulations.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line AuthenticationContamination: All the cell lines tested negative for mycoplasma contamination.
    Authentication: All cell lines used have been authenticated.

    Table 2: Resources

    Antibodies
    SentencesResources
    Primary antibodies were diluted with blocking solution and incubated with cells overnight at 4°C: Cleaved caspase-3 (rabbit, Cell Signaling #9661, 1:500), Anti-Nestin (mouse, Anti-Nestin antibody [10C2]; 1:1000), Anti-Ki67 antibody (rabbit, Abcam ab15580, 1:300), Anti-MAP2 antibody (Chicken, Abcam ab5392, 1:1000), Anti-VGlUT1 [317D4] (mouse, Synaptic Systems #135311, 1:500), Anti Homer 1 (VesL 1, Syn 47) (rabbit, Synaptic Systems #160003, 1:500), anti-GFAP (chicken, Abcam ab4674, 1:1000).
    Cleaved caspase-3
    suggested: (Cell Signaling Technology Cat# 9661, RRID:AB_2341188)
    rabbit, Cell Signaling #9661
    suggested: (Cell Signaling Technology Cat# 9661, RRID:AB_2341188)
    Anti-Nestin
    suggested: (LSBio (LifeSpan Cat# LS-C124240-1000, RRID:AB_10805379)
    Anti-Ki67
    suggested: (Abcam Cat# ab15580, RRID:AB_443209)
    Anti-MAP2
    suggested: (Abcam Cat# ab5392, RRID:AB_2138153)
    Anti-VGlUT1 [317D4]
    suggested: None
    Anti Homer 1 (VesL 1
    suggested: None
    anti-GFAP
    suggested: (Abcam Cat# ab4674, RRID:AB_304558)
    SARS-CoV-2 (2019-nCoV) Nucleoprotein / NP Antibody (rabbit Mab, Sino Biological #40143-R019, 1: 2000) was incubated diluted with blocking solution and incubated with cells for 20 minutes at room temperature.
    SARS-CoV-2 (2019-nCoV) Nucleoprotein / NP
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    The virus was propagated in Vero E6 cells (ATCC® CRL-1586TM) transfected with exogenous human ACE2 and TMPRSS2.
    Vero E6
    suggested: None
    Software and Algorithms
    SentencesResources
    Neuronal Maturation step was achieved by changing the media to Neurobasal with GlutaMAX, 1% Gem21 NeuroPlex (Gemini Bio), 1% NEAA and 1% PS; supplemented with 10 ng/mL of BDNF, 10 ng/mL of GDNF, 10 ng/mL of NT-3 (PeproTech), 200 mM L-ascorbic acid and 1 mM dibutyryl-cAMP (Sigma-Aldrich).
    NeuroPlex
    suggested: (NeuroPlex, RRID:SCR_016193)
    Images were blindly collected using an Axio Observer Z1 Microscope with ApoTome (Zeiss) and analyzed with ImageJ software.
    ImageJ
    suggested: (ImageJ, RRID:SCR_003070)
    Statistical analyses: Results were analyzed using Prism Software (version 6, GraphPad, USA).
    Prism
    suggested: (PRISM, RRID:SCR_005375)
    GraphPad
    suggested: (GraphPad Prism, RRID:SCR_002798)

    Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

    Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

    Results from TrialIdentifier: No clinical trial numbers were referenced.

    Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

    Results from JetFighter: We did not find any issues relating to colormaps.

    Results from rtransparent:
    • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
    • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
    • No protocol registration statement was detected.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.

  2. Version published to 10.1101/2020.05.30.125856 on bioRxiv