Topoisomerase 1 inhibition therapy protects against SARS-CoV-2-induced inflammation and death in animal models

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Abstract

The ongoing pandemic caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is currently affecting millions of lives worldwide. Large retrospective studies indicate that an elevated level of inflammatory cytokines and pro-inflammatory factors are associated with both increased disease severity and mortality. Here, using multidimensional epigenetic, transcriptional, in vitro and in vivo analyses, we report that Topoisomerase 1 (Top1) inhibition suppresses lethal inflammation induced by SARS-CoV-2. Therapeutic treatment with two doses of Topotecan (TPT), a FDA-approved Top1 inhibitor, suppresses infection-induced inflammation in hamsters. TPT treatment as late as four days post-infection reduces morbidity and rescues mortality in a transgenic mouse model. These results support the potential of Top1 inhibition as an effective host-directed therapy against severe SARS-CoV-2 infection. TPT and its derivatives are inexpensive clinical-grade inhibitors available in most countries. Clinical trials are needed to evaluate the efficacy of repurposing Top1 inhibitors for COVID-19 in humans.

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  1. SciScore for 10.1101/2020.12.01.404483: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIACUC: For experiments shown in Figure S2, infection procedures were performed following protocols approved by the Icahn School of Medicine at Mount Sinai Institutional Animal Care and Use Committee (IACUC).
    RandomizationA number was randomly assigned by the investigator to discriminate each section, which was then submitted for analysis.
    BlindingMicroscopic sections were analyzed in a blinded fashion by the same pathologist (A.M.).
    Power Analysisnot detected.
    Sex as a biological variable7-10 week old (~120-140g) female Golden Syrian hamsters (Charles River) were anesthetized using 90mg/kg Ketamine and 2mg/kg Xylazine by intraperitoneal injection.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Each sonicated lysate was then pre-cleared using 10ul of Rabbit-IgG Dynabeads for 1 hour, rotating at 4°C. 1ug of anti-H3K27ac antibody was then added to 300ul of pre-cleared lysate.
    anti-H3K27ac
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    A549 cells were then transduced with the lentivirus in the presence of polybrene (8 μg/ml).
    A549
    suggested: NCI-DTP Cat# A549, RRID:CVCL_0023)
    Preparation of siTOP1 sequencing libraries: 7.5E4 A549-ACE2 cells were plated in a 24 well dishes.
    A549-ACE2
    suggested: None
    Experimental Models: Organisms/Strains
    SentencesResources
    5-10 week old female B6.Cg-Tg(K18-ACE2)2Prlmn/J (K18-hACE2) mice purchased from Jackson Laboratories (Bar Harbor, ME) were anesthetized by an intraperitoneal injection of 90mg/kg Ketamine and 2 mg/kg xylazine.
    B6.Cg-Tg(K18-ACE2)2Prlmn/J (K18-hACE2)
    suggested: None
    Extraction of RNA from Lungs of Infected Hamsters and Mice: Upon euthanasia, the superior lobe of infected hACE2-KI mice or Golden Syrian hamsters were collected for RNA extraction.
    hACE2-KI
    suggested: None
    Software and Algorithms
    SentencesResources
    Images were with Olympus BX53 semi-motorized fluorescence microscope using cellSens imaging software.
    cellSens
    suggested: None

    Results from OddPub: Thank you for sharing your code.


    Results from LimitationRecognizer: We detected the following sentences addressing limitations in the study:
    Pharmacokinetics considerations and limitations of this study: Although the preclinical animal models of SARS-CoV-2 pathogenesis used here are, as with any animal model, only partially representative of the biology of humans, our study indicates a promising effect of TPT by suppressing inflammation in COVID-19. Several factors require careful consideration prior to extrapolating these results towards the design of clinical trials of Top1-inhibition therapy for human COVID-19. First, in our animal models, we can suppress inflammation and reduce disease pathology in the lung using 2 doses of Top1 inhibition therapy with TPT at 2mg/kg intraperitoneally. This equates to a 5-fold reduction from typical chemotherapeutic anti-cancer doses in rodent models (Guichard et al., 2001; Houghton et al., 1995; Nemati et al., 2010). In clinical practice, the Top1 inhibitors TPT and Irinotecan have well-characterized pharmacokinetics and toxicity profiles (Kollmannsberger et al., 1999; Mathijssen et al., 2001), albeit in patients without SARS-CoV-2 infection. Doses that are 5-fold lower than those used in the treatment of small-cell lung cancer (TPT)(Rowinsky et al., 1992; von Pawel et al., 1999) and colorectal cancer (irinotecan)(Andre et al., 1999) are expected to cause little to no toxicity, and importantly no risk of neutropenia. This significant dose reduction, together with the wealth of clinical experience in the use of TPT and irinotecan should reassure us about potential concerns over...

    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: Please consider improving the rainbow (“jet”) colormap(s) used on page 35. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.


    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.

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