Aberrant cell migration contributes to defective airway epithelial repair in childhood wheeze
Thomas Iosifidis, Erika N. Sutanto, Alysia G. Buckley, Laura Coleman, Erin E. Gill, Amy H. Lee, Kak-Ming Ling, Jessica Hillas, Kevin Looi, Luke W. Garratt, Kelly M. Martinovich, Nicole C. Shaw, Samuel T. Montgomery, Elizabeth Kicic-Starcevich, Yuliya V. Karpievitch, Peter Le Souëf, Ingrid A. Laing, Shyan Vijayasekaran, Francis J. Lannigan, Paul J. Rigby, Robert E.W. Hancock, Darryl A. Knight, Stephen M. Stick, Anthony Kicic, Western Australian Epithelial Research Program (WAERP), Australian Respiratory Epithelium Consortium (AusREC)
Thomas Iosifidis, Erika N. Sutanto, Alysia G. Buckley, Laura Coleman, Erin E. Gill, Amy H. Lee, Kak-Ming Ling, Jessica Hillas, Kevin Looi, Luke W. Garratt, Kelly M. Martinovich, Nicole C. Shaw, Samuel T. Montgomery, Elizabeth Kicic-Starcevich, Yuliya V. Karpievitch, Peter Le Souëf, Ingrid A. Laing, Shyan Vijayasekaran, Francis J. Lannigan, Paul J. Rigby, Robert E.W. Hancock, Darryl A. Knight, Stephen M. Stick, Anthony Kicic, Western Australian Epithelial Research Program (WAERP), Australian Respiratory Epithelium Consortium (AusREC)
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Research Article Cell biology Pulmonology

Aberrant cell migration contributes to defective airway epithelial repair in childhood wheeze

Abstract

Abnormal wound repair has been observed in the airway epithelium of patients with chronic respiratory diseases, including asthma. Therapies focusing on repairing vulnerable airways, particularly in early life, present a potentially novel treatment strategy. We report defective lower airway epithelial cell repair to strongly associate with common pre–school-aged and school-aged wheezing phenotypes, characterized by aberrant migration patterns and reduced integrin α5β1 expression. Next generation sequencing identified the PI3K/Akt pathway as the top upstream transcriptional regulator of integrin α5β1, where Akt activation enhanced repair and integrin α5β1 expression in primary cultures from children with wheeze. Conversely, inhibition of PI3K/Akt signaling in primary cultures from children without wheeze reduced α5β1 expression and attenuated repair. Importantly, the FDA-approved drug celecoxib — and its non–COX2-inhibiting analogue, dimethyl-celecoxib — stimulated the PI3K/Akt–integrin α5β1 axis and restored airway epithelial repair in cells from children with wheeze. When compared with published clinical data sets, the identified transcriptomic signature was also associated with viral-induced wheeze exacerbations highlighting the clinical potential of such therapy. Collectively, these results identify airway epithelial restitution via targeting the PI3K–integrin α5β1 axis as a potentially novel therapeutic avenue for childhood wheeze and asthma. We propose that the next step in the therapeutic development process should be a proof-of-concept clinical trial, since relevant animal models to test the crucial underlying premise are unavailable.

Authors

Thomas Iosifidis, Erika N. Sutanto, Alysia G. Buckley, Laura Coleman, Erin E. Gill, Amy H. Lee, Kak-Ming Ling, Jessica Hillas, Kevin Looi, Luke W. Garratt, Kelly M. Martinovich, Nicole C. Shaw, Samuel T. Montgomery, Elizabeth Kicic-Starcevich, Yuliya V. Karpievitch, Peter Le Souëf, Ingrid A. Laing, Shyan Vijayasekaran, Francis J. Lannigan, Paul J. Rigby, Robert E.W. Hancock, Darryl A. Knight, Stephen M. Stick, Anthony Kicic, Western Australian Epithelial Research Program (WAERP), Australian Respiratory Epithelium Consortium (AusREC)

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Figure 2

Role of integrins in pAEC from children with and without wheeze.

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Role of integrins in pAEC from children with and without wheeze. (A and ...
(A and B) Protein expression of integrin subunits α5 (A) and β1 (B) were found to be significantly lower in pAEC from children with wheeze compared with their nonwheezing counterparts. (C–E) Strong immunofluorescence staining of both integrin α5 (C) and β1 (D) on leading edge pAEC from children without wheeze was demonstrated that was almost undetectable in sites distal to the wound (E). (F–H) In contrast, pAEC cultures of children with wheeze exhibited weak integrin α5 (F) and β1 (G) staining along the leading edge cells (H). The slides were counterstained with Hoechst nuclear stain (blue). Representative images of samples from 5 children with wheeze and 5 children without wheeze (C–H). Oil immersion objective, optical magnification of 600×, imaged using 60× objective and 10× ocular lens; NA 1.4. Scale bar: 50 μm. (I) Blocking β1 integrin (1:40 dilution, IgG1κ, P5D2) function significantly reduced pAEC wound closure rates to similar rates observed in their wheezing counterparts (red dashed line). Matching dilution of isotype control antibody (IgG1κ, MOPC-21) or untreated pAEC from children without wheeze reached full closure by 72 hours after wounding. n = 4 children. Median ± IQR, *P < 0.050, Mann-Whitney U test. (J) Cultures treated with isotype antibody (1:40 dilution) displayed comparable migration patterns with untreated pAEC from children without wheeze. (K) However, cultures treated with 1:40 dilution of anti–β1 integrin antibody in culture media migrated less far into the wound, lacking cell directionality and specificity toward the wound center. Individual cell tracks were transposed so that each track had its start at the origin. n = 60 tracks from 4 children without wheeze (I–K).