Rho-Kinase 1/2 Inhibition Prevents Transforming Growth Factor-β-Induced Effects on Pulmonary Remodeling and Repair

Abstract

Transforming growth factor (TGF)-β-induced myofibroblast transformation and alterations in mesenchymal-epithelial interactions contribute to chronic lung diseases such as chronic obstructive pulmonary disease (COPD), asthma and pulmonary fibrosis. Rho-associated coiled-coil-forming protein kinase (ROCK) consists as two isoforms, ROCK1 and ROCK2, and both are playing critical roles in many cellular responses to injury. In this study, we aimed to elucidate the differential role of ROCK isoforms on TGF-β signaling in lung fibrosis and repair. For this purpose, we tested the effect of a non-selective ROCK 1 and 2 inhibitor (compound 31) and a selective ROCK2 inhibitor (compound A11) in inhibiting TGF-β-induced remodeling in lung fibroblasts and slices; and dysfunctional epithelial-progenitor interactions in lung organoids. Here, we demonstrated that the inhibition of ROCK1/2 with compound 31 represses TGF-β-driven actin remodeling as well as extracellular matrix deposition in lung fibroblasts and PCLS, whereas selective ROCK2 inhibition with compound A11 did not. Furthermore, the TGF-β induced inhibition of organoid formation was functionally restored in a concentration-dependent manner by both dual ROCK 1 and 2 inhibition and selective ROCK2 inhibition. We conclude that dual pharmacological inhibition of ROCK 1 and 2 counteracts TGF-β induced effects on remodeling and alveolar epithelial progenitor function, suggesting this to be a promising therapeutic approach for respiratory diseases associated with fibrosis and defective lung repair.

A transcriptomics-guided drug target discovery strategy identifies receptor ligands for lung regeneration

“…The primary alveolar epithelial cells, brief in Epcam + cells (CD31 − /CD45 − /CD326 + ) were isolated on the basis of the protocols published previously from our group ( 67 ). The isolation of Epcam + cells generated from the chronic CS mice was slightly different.…”

The Novel Small Molecule BTB Inhibits Pro-Fibrotic Fibroblast Behavior though Inhibition of RhoA Activity

“…We next identified the ROCK/Rho signaling axis as a promising mechanism given that this pathway has been identified to induce matrix production [ 29 , 30 , 31 ], induce proliferation [ 32 , 33 , 34 , 35 ], and induce myofibroblast differentiation in vitro [ 36 , 37 , 38 ]. Importantly, inhibition of ROCK/Rho signaling inhibits fibrosis in vivo [ 39 , 40 , 41 , 42 , 43 ]. ROCK1 is the most well studied effector in this pathway and is known to be pro-fibrotic in human lug fibroblasts.…”

Application and Study of ROCK Inhibitors in Pulmonary Fibrosis: Recent Developments and Future Perspectives

“…Interestingly, ROCK inhibitors prevent TGF-β-induced lung remodeling and repair, which is attributed to the inhibition of actin reconstruction, lung fibroblast activation, and ECM deposition. 63 Moreover, suppression of ROCK activity led to the polarization of M2-like macrophages, resulting in the attenuation of fibrosis mediated by myofibroblasts. 64 These findings suggest that selective inhibition of either ROCK isoform shows the potential to be an effective strategy for PF treatment.…”