AccScience Publishing / JCTR / Online First / DOI: 10.36922/JCTR025270036
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ORIGINAL ARTICLE

Comparative transcriptomic analysis of macrophages treated with a combination of ROCK pathway inhibitors

Arijita Subuddhi1,2,3 ,  Marta Halasa1,2 ,  Ahmed Uosef1,2 ,  Dawei Zou1 ,  Souhail A. Thabet2 ,  Henry V. Ubelaker1,2 ,  Rafik M. Ghobrial1,2 ,  Malgorzata Kloc1,2*
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1 Immunobiology and Transplant Science Center, Houston Methodist Research Institute, Houston, Texas, United States of America
2 Department of Surgery, Houston Methodist Hospital, Houston, Texas, United States of America
3 Emory/GA Tuberculosis Research Advancement Center (TRAC), Emory Vaccine Center, Atlanta, Georgia, United States of America
JCTR, 025270036 https://doi.org/10.36922/JCTR025270036
Received: 4 July 2025 | Revised: 9 August 2025 | Accepted: 15 August 2025 | Published online: 14 October 2025
© 2025 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution -Noncommercial 4.0 International License (CC-by the license) ( https://creativecommons.org/licenses/by-nc/4.0/ )
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Abstract

Background: At present, there is no therapy for the long-term (chronic) rejection of transplanted organs. This condition leads to tissue fibrosis and occlusion of the blood vessels. Aim: The overall goal of the current research is to identify a clinically applicable therapy for chronic rejection in transplanted organs. Our previous study showed that inhibitors of the RhoA/Rock pathway, such as Rezurock and fingolimod, prevent chronic rejection in rodent transplantation models, with Rezurock being superior in reducing fibrosis. Materials and methods: In this study, we analyzed the effect of a Rezurock and fingolimod combination on the transcriptome of mouse peritoneal macrophages and protein expression in both mouse and human macrophages. Results: The Rezurock/fingolimod combination resulted in the differential expression of 4,855 genes (2,477 downregulated and 2378 upregulated). Downregulated genes were related to fibrotic pathways, extracellular matrix, blood vessel development, cell adhesion, and cytokine production. Protein expression analysis showed that Rezurock/fingolimod treatment had a significantly stronger effect on the expression of pentraxin 3, chemokine (C-C motif) ligand 2, C-C motif chemokine receptor 2, and transforming growth factor beta 1 in mouse macrophages, and was much more effective in reducing the expression of Notch1 and Rho-associated coiled-coil kinase 2 in human macrophages compared to individual treatments. Conclusion: Rezurock/fingolimod treatment not only affects fibrotic pathways but also downregulates genes related to cell cycle progression and cytokine production and disrupts macrophage recruitment signaling. These findings indicate that Rezurock, alone or in combination with other immunomodulators, may be a promising candidate for clinical therapy targeting chronic rejection.

Graphical abstract
Keywords
Rezurock
Fibrosis
Chronic rejection
Macrophage
Ras homolog family member A/Rho-associated coiled-coil kinase pathway
Funding
None.
Conflict of interest
Malgorzata Kloc is Editor-in-Chief of this journal, but was not involved in any way, directly or indirectly, in the editorial and peer-review process for this paper. All other authors declare that they have no known competing financial interests or personal relationships that could have influenced the work reported in this paper.
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Journal of Clinical and Translational Research, Electronic ISSN: 2424-810X Print ISSN: 2382-6533, Published by AccScience Publishing
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