Exploring the Antifibrotic Potential of the heparan sulfate mimetic OTR4120: Insights from Preclinical Models
Manuela Marega1,2,3, Najet Mejdoubi-Charef4 , David Wiegard1, Muzamil Majid Khan5,6, Marek Bartkuhn2, Laura Gambs2, Tara Procida-Kowalski2, Jochem Wilhelm2, Esmeralda Vasquez-Pacheco1, Afshin Noori1,2, Ying Dong1,2, Yi Zheng7, Xuran Chu7, Arun Lingampally1,2, Joanna Zukowska5,6, Said Charef8,9, Franck Chiappini8,9, Agnes Choppin9, Dulce Papy Garcia8, Rainer Pepperkok5,6, Thomas Muley5,10, Hauke Winter5,10, Clemens Ruppert1,2,11, Werner Seeger1,2, Andreas Gunther1,2,11, Denis Barritault9, Chengshui Chen12#, Cho-Ming Chao1,3#, Stefano Rivetti1,2#, Saverio Bellusci12,13#
Abstract
Rationale: Idiopathic pulmonary fibrosis (IPF) is a debilitating lung disease characterized by excessive deposition of extracellular matrix (ECM), resulting in lung function impairment. Heparan sulfate mimetics (HSm) have been suggested to have potential antifibrotic effects by regulating ECM. This study aims to investigate the impact of a specific HSm named OTR4120 on fibrotic processes in ex vivo, in vitro and in vivo models.
Methods: Human Precision Cut Lung Slices (hPCLS) treated with a fibrotic cocktail alone or with OTR4120 were evaluated using second-harmonic imaging microscopy (SHIM) to assess collagen deposition. Human embryonic fibroblast WI-38 cell line and primary fibroblasts obtained from human donors were differentiated into myofibroblasts (MYF) using TGF-β1 and treated with OTR4120 or a control vehicle. Gene expression analysis for MYF markers was performed using quantitative PCR (qPCR). Protein expression of MYF markers was evaluated using immunofluorescence techniques. Bulk-RNA sequencing analysis on WI-38 cells cultured under different experimental conditions was conducted. Finally, the therapeutic effects of OTR4120 on a bleomycin-induced fibrosis mouse model were investigated.
Results: SHIM analysis on OTR4120-treated hPCLS showed a decrease in collagen deposition. OTR4120 treatment of primary fibroblasts and WI-38 cells exposed to
TGF-β1 significantly reduced the expression of MYF markers. Bulk-RNA sequencing analysis on OTR4120-treated WI-38 cells showed significant impacts on fibrosisrelated processes. Therapeutic application of OTR4120 in vivo to bleomycin-induced fibrosis mice resulted in enhanced fibrosis resolution.
Conclusion: OTR4120 has potential therapeutic benefits as an antifibrotic agent in the context of lung fibrosis. Further investigations are necessary to understand the precise mechanism through which OTR4120 exerts its antifibrotic effects.
