Plasma proteomics can discriminate isolated early from dual responses in asthmatic individuals undergoing an allergen inhalation challenge

Authors

  • Amrit Singh,

    1. James Hogg Research Centre, St. Paul's Hospital, University of British Columbia, Vancouver, Canada
    2. Institute for HEART+LUNG Health, Vancouver, Canada
    3. NCE CECR PROOF Centre of Excellence, Vancouver, Canada
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  • Gabriela V. Cohen Freue,

    1. Institute for HEART+LUNG Health, Vancouver, Canada
    2. NCE CECR PROOF Centre of Excellence, Vancouver, Canada
    3. Department of Statistics, Vancouver, Canada
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  • Jean L. Oosthuizen,

    1. James Hogg Research Centre, St. Paul's Hospital, University of British Columbia, Vancouver, Canada
    2. Institute for HEART+LUNG Health, Vancouver, Canada
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  • Sarah H. Y. Kam,

    1. James Hogg Research Centre, St. Paul's Hospital, University of British Columbia, Vancouver, Canada
    2. Institute for HEART+LUNG Health, Vancouver, Canada
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  • Jian Ruan,

    1. James Hogg Research Centre, St. Paul's Hospital, University of British Columbia, Vancouver, Canada
    2. Institute for HEART+LUNG Health, Vancouver, Canada
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  • Mandeep K. Takhar,

    1. Institute for HEART+LUNG Health, Vancouver, Canada
    2. NCE CECR PROOF Centre of Excellence, Vancouver, Canada
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  • Gail M. Gauvreau,

    1. Department of Medicine, McMaster University, Hamilton, Canada
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  • Paul M. O'Byrne,

    1. Department of Medicine, McMaster University, Hamilton, Canada
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  • J. Mark FitzGerald,

    1. Institute for HEART+LUNG Health, Vancouver, Canada
    2. Department of Medicine, Division of Respiratory Medicine, Vancouver, Canada
    3. Vancouver Coastal Health Research Institute, Vancouver General Hospital, Vancouver, Canada
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  • Louis-Philippe Boulet,

    1. Centre de Pneumologie de L'Hopital, Université Laval, Sainte-Foy, Canada
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  • Christoph H. Borchers,

    1. University of Victoria – Genome BC Proteomics Centre, University of Victoria, Victoria, Canada
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  • Scott J. Tebbutt

    Corresponding author
    1. Institute for HEART+LUNG Health, Vancouver, Canada
    2. NCE CECR PROOF Centre of Excellence, Vancouver, Canada
    3. Department of Medicine, Division of Respiratory Medicine, Vancouver, Canada
    • James Hogg Research Centre, St. Paul's Hospital, University of British Columbia, Vancouver, Canada
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  • Colour Online: See the article online to view Figs. 1‒3 in colour.

Correspondence: Dr. Scott J. Tebbutt, UBC James Hogg Research Centre, Room 166, St. Paul's Hospital, 1081 Burrard Street, Vancouver, BC V6Z 1Y6, Canada

E-mail: Scott.Tebbutt@hli.ubc.ca

Fax: 604-806-9274

Abstract

Purpose

This proteomics study was designed to determine the utility of iTRAQ MALDI-TOF/TOF technology to compare plasma samples from carefully phenotyped mild, atopic asthma subjects undergoing allergen inhalation challenge.

Experimental design

Eight adult subjects with mild, allergic asthma (four early responders (ERs) and four dual responders (DRs)) participated in the allergen inhalation challenge. Blood samples were collected prior to and 2 h after the inhalation challenge. Sixteen plasma samples (two per subject), technical replicates, and pooled controls were analyzed using iTRAQ. Technical validation was performed using LC-MRM/MS. Moderated robust regression was used to determine differentially expressed proteins.

Results

Although this study did not show significant differences between pre- and post-challenge samples, discriminant analysis indicated that certain proteins responded differentially to allergen challenge with respect to responder type. At pre-challenge, fibronectin was significantly elevated in DRs compared to ERs and remained significant in the multiple reaction monitoring validation.

Conclusions and clinical relevance

This proof of principle demonstration has shown that iTRAQ can uncover differences in the human plasma proteome between two endotypes of asthma and merits further application of iTRAQ to larger cohorts of asthma and other respiratory diseases.

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