Cystic fibrosis (CF) is an inherited disease associated with chronic, severe lung inflammation, leading to premature death. To develop innovative anti-inflammatory treatments, we need to characterize new cellular and molecular components contributing to the mechanisms of lung inflammation. Here, we focused on the potential role of "transient receptor potential vanilloid-4" (TRPV4), a non-selective, calcium channel. We first showed that 5,6-, 8,9-, 11,12- and 14,15-epoxyeicosatrienoic acids, i.e. four natural lipid-based TRPV4 agonists, are present in expectorations of CF patients. We also used both in vitro and in vivo approaches to demonstrate that TRPV4 expressed in airway epithelial cells triggers the secretion of major pro-inflammatory mediators such as chemokines and biologically active lipids, as well as a neutrophil recruitment into lung tissues. We further characterized the contribution of cytosolic phospholipase A2, MAPKs and NF-B in TRPV4-dependent signaling. Finally, we observed an alteration of TRPV4-induced inflammatory response in a CF context, suggesting that TRPV4 is a promising target for the development of new anti-inflammatory treatments for diseases such as CF.