Overriding impaired FPR chemotaxis signaling in diabetic neutrophil stimulates infection control in murine diabetic wound

  1. Ruchi Roy
  2. Janet Zayas
  3. Sunil K Singh
  4. Kaylee Delgado
  5. Stephen J Wood
  6. Mohamed F Mohamed
  7. Dulce M Frausto
  8. Ricardo Estupinian
  9. Eileena F Giurini
  10. Timothy M Kuzel
  11. Andrew Zloza
  12. Jochen Reiser
  13. Sasha H Shafikhani  Is a corresponding author
  1. Rush University Medical Center, United States
  2. University of Illinois at Chicago, United States

Abstract

Infection is a major co-morbidity that contributes to impaired healing in diabetic wounds. Although impairments in diabetic neutrophils have been blamed for this co-morbidity, what causes these impairments and whether they can be overcome, remain largely unclear. Diabetic neutrophils, isolated from diabetic individuals, exhibit chemotaxis impairment but this peculiar functional impairment has been largely ignored because it appears to contradict the clinical findings which blame excessive neutrophil influx as a major impediment to healing in chronic diabetic ulcers. Here, we report that exposure to glucose in diabetic range results in impaired chemotaxis signaling through the formyl peptide receptor (FPR) in neutrophils, culminating in reduced chemotaxis and delayed neutrophil trafficking in the wound of Leprdb (db/db) type 2 diabetic mice, rendering diabetic wound vulnerable to infection. We further show that at least some auxiliary receptors remain functional under diabetic conditions and their engagement by the pro-inflammatory cytokine CCL3, overrides the requirement for FPR signaling and substantially improves infection control by jumpstarting the neutrophil trafficking toward infection, and stimulates healing in diabetic wound. We posit that CCL3 may have therapeutic potential for the treatment of diabetic foot ulcers if it is applied topically after the surgical debridement process which is intended to reset chronic ulcers into acute fresh wounds.

Data availability

All data generated or analyzed during this study are included in the manuscript and supporting files. Source data files have been provided for western blots.

Article and author information

Author details

  1. Ruchi Roy

    Department of Medicine, Rush University Medical Center, Chicago, United States
    Competing interests
    No competing interests declared.
  2. Janet Zayas

    Department of Medicine, Rush University Medical Center, Chicago, United States
    Competing interests
    No competing interests declared.
  3. Sunil K Singh

    Department of Surgery, Division of Surgical Oncology, University of Illinois at Chicago, Chicago, United States
    Competing interests
    No competing interests declared.
  4. Kaylee Delgado

    Department of Medicine, Rush University Medical Center, Chicago, United States
    Competing interests
    No competing interests declared.
  5. Stephen J Wood

    Department of Microbial Pathogens and Immunity, Rush University Medical Center, Chicago, United States
    Competing interests
    No competing interests declared.
  6. Mohamed F Mohamed

    Department of Medicine, Rush University Medical Center, Chicago, United States
    Competing interests
    No competing interests declared.
  7. Dulce M Frausto

    Department of Medicine, Rush University Medical Center, Chicago, United States
    Competing interests
    No competing interests declared.
  8. Ricardo Estupinian

    Department of Medicine, Rush University Medical Center, Chicago, United States
    Competing interests
    No competing interests declared.
  9. Eileena F Giurini

    Department of Medicine, Rush University Medical Center, Chicago, United States
    Competing interests
    No competing interests declared.
  10. Timothy M Kuzel

    Department of Medicine, Rush University Medical Center, Chicago, United States
    Competing interests
    No competing interests declared.
  11. Andrew Zloza

    Department of Medicine, Rush University Medical Center, Chicago, United States
    Competing interests
    No competing interests declared.
  12. Jochen Reiser

    Department of Medicine, Rush University Medical Center, Chicago, United States
    Competing interests
    No competing interests declared.
  13. Sasha H Shafikhani

    Department of Medicine, Rush University Medical Center, Chicago, United States
    For correspondence
    Sasha_Shafikhani@rush.edu
    Competing interests
    Sasha H Shafikhani, Rush University Medical Center has filed a patent (International Application Number: PCT/US19/41112). Dr. Sasha Shafikhani is the listed inventor on this application..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1755-9997

Funding

National Institutes of Health (RO1DK107713)

  • Sasha H Shafikhani

National Institutes of Health (R01AI150668)

  • Sasha H Shafikhani

National Institutes of Health (F31DK118797)

  • Janet Zayas

National Institutes of Health (GM109421)

  • Janet Zayas

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: We have an approval from the Rush University Medical Center Institutional Animal Care and Use Committee (IACUC No.: 18-037) to conduct research as indicated. All procedures complied strictly with the standards for care and use of animal subjects as stated in the Guide for the Care and Use of Laboratory Animals (Institute of Laboratory Animal Resources, National Academy of Sciences, Bethesda, MD, USA).

Human subjects: We have an Institutional Review Board (IRB)- approved protocol in accordance with the Common Rule (45CFR46, December 13, 2001) and any other governing regulations or subparts. This IRB-approved protocol allows us to collect blood samples from non-diabetic volunteers with their consents for these studies.

Reviewing Editor

  1. Ursula Rescher, University of Muenster, Germany

Publication history

  1. Received: July 9, 2021
  2. Preprint posted: September 11, 2021 (view preprint)
  3. Accepted: February 1, 2022
  4. Accepted Manuscript published: February 3, 2022 (version 1)
  5. Version of Record published: February 15, 2022 (version 2)

Copyright

© 2022, Roy et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 509
    Page views
  • 110
    Downloads
  • 4
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Ruchi Roy
  2. Janet Zayas
  3. Sunil K Singh
  4. Kaylee Delgado
  5. Stephen J Wood
  6. Mohamed F Mohamed
  7. Dulce M Frausto
  8. Ricardo Estupinian
  9. Eileena F Giurini
  10. Timothy M Kuzel
  11. Andrew Zloza
  12. Jochen Reiser
  13. Sasha H Shafikhani
(2022)
Overriding impaired FPR chemotaxis signaling in diabetic neutrophil stimulates infection control in murine diabetic wound
eLife 11:e72071.
https://doi.org/10.7554/eLife.72071
  1. Further reading

Further reading

    1. Immunology and Inflammation
    Lyra O Randzavola, Paige M Mortimer ... David C Thomas
    Research Article

    EROS (Essential for Reactive Oxygen Species) protein is indispensable for expression of gp91phox, the catalytic core of the phagocyte NADPH oxidase. EROS deficiency in humans is a novel cause of the severe immunodeficiency, chronic granulomatous disease (CGD), but its mechanism of action was unknown until now. We elucidate the role of EROS, showing it acts at the earliest stages of gp91phox maturation. It binds the immature 58kDa gp91phox directly, preventing gp91phox degradation and allowing glycosylation via the oligosaccharyltransferase (OST) machinery and the incorporation of the heme prosthetic groups essential for catalysis. EROS also regulates the purine receptors P2X7 and P2X1 through direct interactions and P2X7 is almost absent in EROS deficient mouse and human primary cells. Accordingly, lack of murine EROS results in markedly abnormal P2X7 signalling, inflammasome activation and T cell responses. The loss of both ROS and P2X7 signalling leads to resistance to influenza infection in mice. Our work identifies EROS as a highly selective chaperone for key proteins in innate and adaptive immunity and a rheostat for immunity to infection. It has profound implications for our understanding of immune physiology, ROS dysregulation and possibly gene therapy.

    1. Computational and Systems Biology
    2. Immunology and Inflammation
    Mingyao Pan, Bo Li
    Short Report Updated

    T cells are potent at eliminating pathogens and playing a crucial role in the adaptive immune response. T cell receptor (TCR) convergence describes T cells that share identical TCRs with the same amino acid sequences but have different DNA sequences due to codon degeneracy. We conducted a systematic investigation of TCR convergence using single-cell immune profiling and bulk TCRβ-sequence (TCR-seq) data obtained from both mouse and human samples and uncovered a strong link between antigen-specificity and convergence. This association was stronger than T cell expansion, a putative indicator of antigen-specific T cells. By using flow-sorted tetramer+ single T cell data, we discovered that convergent T cells were enriched for a neoantigen-specific CD8+ effector phenotype in the tumor microenvironment. Moreover, TCR convergence demonstrated better prediction accuracy for immunotherapy response than the existing TCR repertoire indexes. In conclusion, convergent T cells are likely to be antigen-specific and might be a novel prognostic biomarker for anti-cancer immunotherapy.