View Abstract

A7653 - Drug Repurposing for WNT/β-Catenin Induced Lung Repair in Chronic Obstructive Pulmonary Disease
Author Block: R. Costa1, D. E. Wagner1, M. M. De Santis1, K. E. Bailey2, A. Doryab1, K. Schorpp3, I. Rothenaigner3, C. Ota1, H. A. Baarsma1, M. Campillos4, K. Hadian3, M. Koenigshoff2; 1Comprehensive Pneumology Center (CPC), Helmholtz Zentrum München and University Hospital of the Ludwig Maximilians University, Member of the German Center for Lung Research (DZL), Munich, Germany, 2University of Colorado, School of Medicine, Division of Pulmonary Sciences and Critical Care Medicine, Aurora, CO, United States, 3Institute of Molecular Toxicology and Pharmacology, Assay Development and Screening Platform, Helmholtz Zentrum München, Munich, Germany, 4Institute of Bioinformatics and Systems Biology, Systems Biology of Small Molecules, Helmholtz Zentrum München, Munich, Germany.
Rationale: Chronic obstructive pulmonary disease (COPD) is an incurable lung disease characterized by inflammation and progressive loss of tissue (emphysema). Importantly, WNT/β-catenin signaling is reduced in patients with COPD. It could be demonstrated in previous reports from our laboratory that pharmacological activation of the pathway via glycogen synthase kinase 3 (GSK3) inhibition decreased emphysematous markers (e.g. elastin and MMP12) in a mouse model of emphysema in vivo and in 3D human and mouse lung tissue slices ex vivo. Here, we aimed to identify new pharmaceutical compounds which activate WNT/β-catenin signaling and therefore induce lung repair.
Methods: We performed a high-throughput drug screen, using an NIH3T3 cell line stably expressing luciferase under the control of WNT/β-catenin responsive promoters (TCF/LEF). 1,216 Food and Drug Administration (FDA) approved drugs (Prestwick Chemical library) and 30,000 novel compounds, belonging to small-molecule diversity libraries were tested. Using the target prediction tool HitPick, we predicted the protein targets for each hit. We selected candidate hits which had a predicted protein target associated with the functional annotations “epithelial cell proliferation”, “regulation of inflammatory responses” and with the keywords “WNT” and “COPD”. Next, we confirmed our initial results by performing TOP/FOP luciferase and an ATP based luminescent cell viability assay. We further investigated the regenerative potential of the candidate drugs using primary mouse alveolar type II cell-derived organoids.
Results: In our primary screening we have identified 652 compounds, activating β-catenin signaling. Sixteen of these are FDA approved drugs (e.g. Riluzole). We predicted protein targets for 154 of the primary hits and selected five compounds for further investigation. We could confirm that all five of the selected candidate compounds induced luciferase activity in a dose dependent manner without inducing cell proliferation. Preliminary experiments indicate that the selected compounds are able to induce organoid formation after 14 days of treatment.
Conclusion: We identified several compounds capable of activating WNT/β-catenin signaling, which may be used to develop potential novel therapies for COPD patients. Drug repurposing of our five candidate FDA approved candidate compounds is of special interest for expediting potential translation into the clinic.