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The Journal of Heart and Lung Transplantation
International Society for Heart and Lung Transplantation.

The mitigating effect of exogenous carbon monoxide on chronic allograft rejection and fibrosis post-lung transplantation

Published:November 25, 2022DOI:https://doi.org/10.1016/j.healun.2022.11.005

      Background

      Small airway inflammation and fibrosis or bronchiolitis obliterans (BO) is the predominant presentation of chronic lung allograft dysfunction (CLAD) post-lung transplantation. Carbon monoxide (CO) is a critical endogenous signaling transducer with known anti-inflammatory and anti-fibrotic effects but its therapeutic potential in CLAD remains to be fully elucidated.

      Methods

      Here we investigate the effect of inhaled CO in modulating chronic lung allograft rejection pathology in a murine orthotopic lung transplant model of BO (B6D2F1/J→DBA/2J). Additionally, the effects of CO on the activated phenotype of mesenchymal cells isolated from human lung transplant recipients with CLAD were studied.

      Results

      Murine lung allografts treated with CO (250 ppm × 30 minutes twice daily from days 7 to 40 post-transplantation) demonstrated decreased immune cell infiltration, fibrosis, and airway obliteration by flow cytometry, trichrome staining, and morphometric analysis, respectively. Decreased total collagen, with levels comparable to isografts, was noted in CO-treated allografts by quantitative hydroxyproline assay. In vitro, CO (250 ppm × 16h) was effective in reversing the fibrotic phenotype of human CLAD mesenchymal cells with decreased collagen I and β-catenin expression as well as an inhibitory effect on ERK1/2 MAPK, and mTORC1/2 signaling. Sildenafil, a phosphodiesterase 5 inhibitor, partially mimicked the effects of CO on CLAD mesenchymal cells and was partially effective in decreasing collagen deposition in murine allografts, suggesting the contribution of cGMP-dependent and -independent mechanisms in mediating the effect of CO.

      Conclusion

      These results suggest a potential role for CO in alleviating allograft fibrosis and mitigating chronic rejection pathology post-lung transplant.

      KEYWORDS

      Abbreviations:

      CO (Carbon Monoxide), CLAD (Chronic Lung Allograft Dysfunction), BO (Bronchiolitis Obliterans), cGMP (cyclic Guanosine Monophosphate), MCs (mesenchymal cells), H&E (Hematoxylin and eosin), PBS (phosphate-buffered saline), mTORC1/2 (mammalian target of rapamycin complex 1/2), 4E-BP1 (Eukaryotic translation initiation factor 4E-binding protein 1), S6K1 (p70 ribosomal protein S6 kinase 1), ERK1/2-MAPK (extracellular signal‑regulated protein kinase1/2-mitogen activated protein kinase), CLAD-MCs (mesenchymal cells from transplant patients diagnosed with CLAD), DMEM (Dulbecco's Minimum Eagle Medium)
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