Cystic fibrosis transmembrane conductance regulator dysfunction in platelets drives lung hyperinflammation
Guadalupe Ortiz-Muñoz, … , Alan S. Verkman, Mark R. Looney
Guadalupe Ortiz-Muñoz, … , Alan S. Verkman, Mark R. Looney
Published January 21, 2020
Citation Information: J Clin Invest. 2020;130(4):2041-2053. https://doi.org/10.1172/JCI129635.
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Research Article Inflammation Pulmonology

Cystic fibrosis transmembrane conductance regulator dysfunction in platelets drives lung hyperinflammation

Abstract

Cystic fibrosis (CF) lung disease is characterized by an inflammatory response that can lead to terminal respiratory failure. The cystic fibrosis transmembrane conductance regulator (CFTR) is mutated in CF, and we hypothesized that dysfunctional CFTR in platelets, which are key participants in immune responses, is a central determinant of CF inflammation. We found that deletion of CFTR in platelets produced exaggerated acute lung inflammation and platelet activation after intratracheal LPS or Pseudomonas aeruginosa challenge. CFTR loss of function in mouse or human platelets resulted in agonist-induced hyperactivation and increased calcium entry into platelets. Inhibition of the transient receptor potential cation channel 6 (TRPC6) reduced platelet activation and calcium flux, and reduced lung injury in CF mice after intratracheal LPS or Pseudomonas aeruginosa challenge. CF subjects receiving CFTR modulator therapy showed partial restoration of CFTR function in platelets, which may be a convenient approach to monitoring biological responses to CFTR modulators. We conclude that CFTR dysfunction in platelets produces aberrant TRPC6-dependent platelet activation, which is a major driver of CF lung inflammation and impaired bacterial clearance. Platelets and TRPC6 are what we believe to be novel therapeutic targets in the treatment of CF lung disease.

Authors

Guadalupe Ortiz-Muñoz, Michelle A. Yu, Emma Lefrançais, Beñat Mallavia, Colin Valet, Jennifer J. Tian, Serena Ranucci, Kristin M. Wang, Zhe Liu, Nicholas Kwaan, Diana Dawson, Mary Ellen Kleinhenz, Fadi T. Khasawneh, Peter M. Haggie, Alan S. Verkman, Mark R. Looney

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Figure 4

Characterization of TRPC6 in platelets.

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Characterization of TRPC6 in platelets. (A, B) mRNA expression of TRPC i...
(A, B) mRNA expression of TRPC isoforms TRPC1 and TRPC6 in platelets from WT, CFTR–/–, CFfl/fl, CF-LysM, and CF-PF4 mice. TRPC isoforms 2, 3, 4, 5, and 7 were undetectable (not shown). (C) Immunofluorescence staining and (D) flow cytometry analysis of CD41 (red) and TRPC6 (blue) in platelets from WT and TRPC6–/– mice (representative of 3 independent experiments). Scale bar: 2.5 μm. (E and F) CD62P expression on platelets from (E) WT, CFTR–/–, TRPC6–/–, and CFTR–/– × TRPC6–/– mice, and (F) CFfl/fl and CF-PF4 mice after thrombin challenge with or without incubation with vehicles, CF172, or CF172 plus SKF-96365 (SK). Data are mean ± SEM of 5 to 11 animals per group. Data were analyzed by 2-way ANOVA. *P ≤ 0.05; **P ≤ 0.01; ****P ≤ 0.0001.