2. Academic Validation
  3. "Keep on ROCKIn": Repurposed ROCK inhibitors to boost corneal endothelial regeneration

"Keep on ROCKIn": Repurposed ROCK inhibitors to boost corneal endothelial regeneration

  • Biomed Pharmacother. 2024 Mar 20:174:116435. doi: 10.1016/j.biopha.2024.116435.
Hendrik Vercammen 1,  Martin Ondra 2,  Jana Kotulova 3,  Edgar Cardenas De La Hoz 4,  Charissa Witters 5,  Katerina Jecmenova 3,  Maxim Le Compte 6,  Christophe Deben 6,  Sorcha Ní Dhubhghaill 7,  Carina Koppen 8,  Marián Hajdúch 2,  Bert Van den Bogerd 9
Affiliations

Affiliations

  • 1 Antwerp Research Group for Ocular Science (ARGOS), Translational Neurosciences, Faculty of Medicine and Health Sciences, University of Antwerp, Wilrijk, Belgium; Department of Ophthalmology, Antwerp University Hospital, Edegem, Belgium; DrugVision Lab, University of Antwerp, Wilrijk, Belgium. Electronic address: hendrik.vercammen@uantwerpen.be.
  • 2 Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University Olomouc, Olomouc, Czech Republic; Czech Advanced Technology and Research Institute (CATRIN), Palacky University Olomouc, Olomouc, Czech Republic.
  • 3 Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University Olomouc, Olomouc, Czech Republic.
  • 4 Industrial Vision Lab, University of Antwerp, Wilrijk, Belgium.
  • 5 Antwerp Research Group for Ocular Science (ARGOS), Translational Neurosciences, Faculty of Medicine and Health Sciences, University of Antwerp, Wilrijk, Belgium; Department of Ophthalmology, Antwerp University Hospital, Edegem, Belgium; DrugVision Lab, University of Antwerp, Wilrijk, Belgium.
  • 6 DrugVision Lab, University of Antwerp, Wilrijk, Belgium.
  • 7 Department of Ophthalmology, Brussels University Hospital, Jette, Belgium.
  • 8 Antwerp Research Group for Ocular Science (ARGOS), Translational Neurosciences, Faculty of Medicine and Health Sciences, University of Antwerp, Wilrijk, Belgium; Department of Ophthalmology, Antwerp University Hospital, Edegem, Belgium.
  • 9 Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University Olomouc, Olomouc, Czech Republic; Czech Advanced Technology and Research Institute (CATRIN), Palacky University Olomouc, Olomouc, Czech Republic. Electronic address: Bert.VandenBogerd@uantwerpen.be.
PMID: 38513591 DOI: 10.1016/j.biopha.2024.116435
Abstract

The global shortage of corneal endothelial graft tissue necessitates the exploration of alternative therapeutic strategies. Rho-associated protein kinase inhibitors (ROCKi), recognized for their regenerative potential in cardiology, oncology, and neurology, have shown promise in corneal endothelial regeneration. This study investigates the repurposing potential of additional ROCKi compounds. Through screening a self-assembled library of ROCKi on B4G12 corneal endothelial cells, we evaluated their dose-dependent effects on proliferation, migration, and toxicity using live-cell imaging. Nine ROCKi candidates significantly enhanced B4G12 proliferation compared to the basal growth rate. These candidates were further assessed for their potential to accelerate wound closure as another indicator for tissue regeneration capacity, with most demonstrating notable efficacy. To assess the potential impact of candidate ROCKi on key corneal endothelial cell markers related to cell proliferation, leaky tight junctions and ion efflux capacity, we analyzed the protein expression of cyclin E1, CDK2, p16, ZO-1 and Na+/K+-ATPase, respectively. Immunocytochemistry and western blot analysis confirmed the preservation of corneal endothelial markers post-treatment with ROCKi hits. However, notable cytoplasm enlargement and nuclear fragmentation were detected after the treatment with SR-3677 and Thiazovivin, indicating possible cellular stress. In compared parameters, Chroman-1 at a concentration of 10 nM outperformed other ROCKi, requiring significantly 1000-fold lower effective concentration than established ROCKi Y-27632 and Fasudil. Altogether, this study underscores the potential of repurposing ROCKi for treating corneal endothelial dysfunctions, offering a viable alternative to conventional grafting methods, and highlights Chroman-1 as a promising candidate structure for hit-to-lead development.

Keywords

Chroman-1; Corneal endothelial regeneration; Drug repurposing; ROCK inhibitors; Small molecule screening.

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