Repurposing of small molecule adrenoreceptor-inhibitor carvedilol for treatment of the fibrotic lung

Maria Jordan^1,2*Kevin Schmidt^1,2Maximilian Fuchs^1,2Annette Just³Angelika Pfanne³Lena Willmer^1,2Lavinia Neubert⁴Christopher Werlein⁴Patrick Zardo⁵Andreas Pich⁶Thomas Thum^3,7Jan Fiedler^1,2*

• ¹Preclinical Pharmacology and Toxicology, Fraunhofer Institute for Toxicology and Experimental Medicine ITEM, Hannover, Germany
• ²Fraunhofer Cluster of Excellence for Immune Mediated Diseases (CIMD), Hannover, Germany
• ³Institute of Molecular and Translational Therapeutic Strategies (IMTTS), Hannover Medical School, Hanover, Lower Saxony, Germany
• ⁴Institute of Pathology, Hannover Medical School, Hannover, Germany
• ⁵Department of Cardiothoracic Transplantation and Vascular Surgery, Hannover Medical School, Hannover, Germany
• ⁶Institute of Toxicology and Core Unit Proteomics, Hannover Medical School, Hannover, Germany
• ⁷Center for Translational Regenerative Medicine, Hannover Medical School, Hannover, Germany

Introduction: Idiopathic pulmonary fibrosis (IPF) is a highly mortal chronic fibrotic lung disease.Current therapies are very limited with nintedanib and pirfenidone being the only non-invasive but also non-curative interventions ultimately bridging to lung transplantation.In silico modelling of dysregulated pathways in IPF and screening for putative interfering small molecules identified carvedilol as a promising anti-fibrotic candidate. We validated drugmediated effects on key features of fibroblast activation in functional assays and gene expression analyses in human embryonic lung fibroblasts (MRC-5). Precision cut lung slices (PCLS) generated from human lung tissue were assessed for secreted fibrotic marker expression.Treatment with carvedilol reduced metabolic activity, inhibited cell proliferation, and led to decreased migratory activity in scratch wound assays in human lung fibroblasts. The functional profile was reflected at the transcriptional level as commonly known fibrotic marker genes, e.g. alpha smooth muscle actin and collagen 1, were robustly repressed. Proteomic profiling underlined a strong extracellular matrix interference with elevated syntheses of several collagen types as well as various integrins, which play a critical role in pro-fibrotic downstream signaling. Comparison of healthy and fibrotic lung tissue validated an upregulation of pro-fibrotic miR-21 secretion in the ex vivo PCLS model which remained unchanged upon carvedilol therapy.Herein, Carvedilol demonstrated significant anti-fibrotic effects on human lung fibroblasts in vitro thus presenting great potential as an anti-IPF treatment. In addition, miR-21 was validated as a secreted pro-fibrotic biomarker in the ex vivo PCLS model.