First-in-human demonstration of splenic ultrasound stimulation for non-invasively controlling inflammation

  1. Rachel S. Graham
  2. Daniel P. Zachs
  3. Victoria Cotero
  4. Catherine D’Agostino
  5. Despoina Ntiloudi
  6. Claire R.W. Kaiser
  7. John Graf
  8. Kirk Wallace
  9. Richard Ramdeo
  10. Thomas R. Coleman
  11. Jeffrey Ashe
  12. John Pellerito
  13. Kevin J. Tracey
  14. Bryce A. Binstadt
  15. Sangeeta S. Chavan
  16. Stavros Zanos
  17. Christopher Puleo
  18. Erik Peterson
  19. Hubert H. Lim

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Abstract

Hyperinflammation and uncontrolled cytokine release in infections and autoimmune diseases require therapy to reduce the innate immune response. Here, we present first in-human data showing reduction in pro-inflammatory cytokine release with ultrasound stimulation of the spleen in healthy subjects and in rheumatoid arthritis patients. Single cell RNA sequencing reveals a decrease in IL-1β and IL-8 transcript levels in circulating monocytes. There is also a down regulation of pathways involved in TNF and IL-6 production, and IFNγ- and NFκB-regulated genes. Additional pre-clinical studies reveal that ultrasound can boost B cell activation and antibody production. Splenic ultrasound offers a new non-invasive therapy for treating hyperinflammation without compromising the adaptive immune response.

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  1. Version published to 10.1101/2020.07.14.20153528v2 on medRxiv
  2. ScreenIT

    SciScore for 10.1101/2020.07.14.20153528: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementConsent: The protocol, informed consent form, recruitment materials, and all participant materials were submitted and approved by the Institutional Review Board (IRB) at Northwell Health.
    IRB: The protocol, informed consent form, recruitment materials, and all participant materials were submitted and approved by the University of Minnesota’s Institutional Review Board (IRB) and monitored by CTSI in accordance with its institutionally approved monitoring plan.
    IACUC: Experiments were performed under protocols approved by the Institutional Animal Care and Use Committee of GE Research.
    RandomizationSplenic ultrasound in RA patients: Design of clinical study: The study is a double-blinded, controlled and randomized clinical trial investigating the therapeutic effects of a 14-day regimen of splenic ultrasound stimulation in RA patients (listed at clinicaltrials.gov (NCT03690466) for which a portion of the results are presented in this paper.
    BlindingA blinded researcher then diluted the LPS to the test concentrations in 15 pre-labeled tubes containing replicates of the different concentrations of LPS used for blood exposure, in which dilutions were made to achieve 0, 0.1, 1, and 10 ng/mL of LPS when mixed with the blood.
    Power Analysisnot detected.
    Sex as a biological variableWomen of childbearing potential were asked to provide a urine sample for pregnancy testing, and approximately 21 mL of blood was drawn.

    Table 2: Resources

    Antibodies
    SentencesResources
    Inclusion and exclusion criteria: To be eligible to participate in the study, an individual must have met the following criteria: 18 years of age or older, a diagnosis of seropositive RA (rheumatoid factor-positive or cyclic citrullinated peptide antibody-positive), and symptoms or signs of inadequate disease control (either modified HAQ score >0.3 or DAS-28-CRP >3.2).
    antibody-positive
    suggested: None
    Serum was tested using ELISA kits for quantifying antibody production including IgG (Abcam, Ab189578), and IgM (Abcam, Ab215085) as per manufacturer’s instructions.
    IgG (Abcam, Ab189578)
    suggested: None
    IgM (Abcam, Ab215085) as per manufacturer’s
    suggested: None
    Experimental Models: Organisms/Strains
    SentencesResources
    Adult Sprague-Dawley rats that were 8-12 weeks old (250-300 g; Charles River Labs) were housed at 25 degrees Celsius on a 12-h light/dark cycle and acclimatized for 1 week, with handling before experiments to minimize the potential confounding measures due to stress response.
    Sprague-Dawley
    suggested: None
    Software and Algorithms
    SentencesResources
    Transcriptomic analysis was performed using R statistical software, and primary filtering and normalization of each sample was done using Seurat package as previously described (30, 59, 60).
    Seurat
    suggested: (SEURAT, RRID:SCR_007322)
    Hs.eg.db and topGO packages.
    topGO
    suggested: (topGO, RRID:SCR_014798)
    Adult Sprague-Dawley rats that were 8-12 weeks old (250-300 g; Charles River Labs) were housed at 25 degrees Celsius on a 12-h light/dark cycle and acclimatized for 1 week, with handling before experiments to minimize the potential confounding measures due to stress response.
    Charles River Labs
    suggested: None
    The single cell RNA sequencing data presented in this paper have been deposited in the Gene Expression Omnibus database as GSExxxxxx (currently being submitted).
    Gene Expression Omnibus
    suggested: (Gene Expression Omnibus (GEO, RRID:SCR_005012)

    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: We found the following clinical trial numbers in your paper:

    IdentifierStatusTitle
    NCT03548116Active, not recruitingBiological Effects of Ultrasound Insonification of the Splee…
    NCT03690466RecruitingUltrasound Treatment of Rheumatoid Arthritis

    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.

  3. Version published to 10.1101/2020.07.14.20153528v1 on medRxiv