IL-13–induced airway mucus production is attenuated by MAPK13 inhibition
Yael G. Alevy, Anand C. Patel, Arthur G. Romero, Dhara A. Patel, Jennifer Tucker, William T. Roswit, Chantel A. Miller, Richard F. Heier, Derek E. Byers, Tom J. Brett, Michael J. Holtzman
Yael G. Alevy, Anand C. Patel, Arthur G. Romero, Dhara A. Patel, Jennifer Tucker, William T. Roswit, Chantel A. Miller, Richard F. Heier, Derek E. Byers, Tom J. Brett, Michael J. Holtzman
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Research Article

IL-13–induced airway mucus production is attenuated by MAPK13 inhibition

Abstract

Increased mucus production is a common cause of morbidity and mortality in inflammatory airway diseases, including asthma, chronic obstructive pulmonary disease (COPD), and cystic fibrosis. However, the precise molecular mechanisms for pathogenic mucus production are largely undetermined. Accordingly, there are no specific and effective anti-mucus therapeutics. Here, we define a signaling pathway from chloride channel calcium-activated 1 (CLCA1) to MAPK13 that is responsible for IL-13–driven mucus production in human airway epithelial cells. The same pathway was also highly activated in the lungs of humans with excess mucus production due to COPD. We further validated the pathway by using structure-based drug design to develop a series of novel MAPK13 inhibitors with nanomolar potency that effectively reduced mucus production in human airway epithelial cells. These results uncover and validate a new pathway for regulating mucus production as well as a corresponding therapeutic approach to mucus overproduction in inflammatory airway diseases.

Authors

Yael G. Alevy, Anand C. Patel, Arthur G. Romero, Dhara A. Patel, Jennifer Tucker, William T. Roswit, Chantel A. Miller, Richard F. Heier, Derek E. Byers, Tom J. Brett, Michael J. Holtzman

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Figure 6

Discovery and validation of potent MAPK13 inhibitors.

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Discovery and validation of potent MAPK13 inhibitors. (A) Superposition ...
(A) Superposition of BIRB-796 into the MAPK13 binding pocket. The position of the gatekeeper Met107 is shown in a space-filling model. Circles indicate ATP-binding and Phe pockets. (B) Crystal structure of compound 61 (cyan) bound to MAPK13 in DFG-out mode. Difference electron density for the compound following rigid-body refinement is shown in mesh. Circles indicate binding pocket features. Phe169 (of the DFG sequence) is highlighted in yellow. (C) Crystal structure of compound 124 (magenta) bound to MAPK13 in DFG-in mode. (D) MAPK13 binding pockets for compound 61 (cyan) and compound 124 (magenta). All residues that make van der Waals contacts or polar contacts are shown. Residues that make direct hydrogen bonds (black lines) with inhibitors are labeled in black. (E) BioLayer interferometry sensorgrams for compound 61 and 124 binding to MAPK13. Processed data are shown as black lines with global kinetic fits overlaid as red lines. (F) MAPK13 inhibitory activity in a screen of test compounds (numbered from 43–205 and arranged from most to least potent) using IMAP assay. Values represent IC50 for compounds with values <20 εM. Red-colored bars indicate compounds selected for detailed analysis. (G) Concentration response for MAPK13 inhibition for indicated test compounds. Values represent fluorescence polarization (mP) from IMAP assay. (H) hTECs were cultured with or without IL-13 for 21 days with the indicated compounds, and MUC5AC levels were determined by ELISA. (G and H) Values represent mean ± SEM. *P < 0.05, decrease from no drug treatment condition.