ROCK2 inhibition has a dual role in reducing ECM remodelling and cell growth, while impairing migration and invasion
Posted on: 27 November 2025
Preprint posted on 2 September 2025
Categories: biophysics, cancer biology, cell biology
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Background and hypothesis:
Key Findings:
ROCK2 inhibition reduces ECM remodelling by fibroblasts
To study the effect of ROCK2 inhibition on ECM remodelling, fibroblasts were embedded in a fibrillar collagen matrix and treated with GV101. It was seen that treatment with GV101 reduces ECM contraction and reduces matrix stiffness in both murine and human models. Treatment with GV101 resulted in reduced formation of highly bundled collagen fibres, indicating that ROCK2 may be required for the formation of mature collagen fibres in murine fibroblasts, while this treatment affected all stages of collagen fibre maturation, bundling, and crosslinking in human CAFs. Furthermore, GV101 also adversely affects the de novo fibrillar production in both murine and human models.
ROCK2 inhibition reduces epithelial cell growth, sensitises cells to shear stress and affects cell migratory dynamics.
Both murine (PyMT) and human (MDA-MB-231) cells exhibit decreased cell viability upon GV101 treatment in 2D, accompanied by reduced colony-forming capacities in 3D. Similarly, an increase in apoptosis following shear stress is observed in both cell types upon ROCK2 inhibition.
Cells were seeded on cell-derived matrices to check the effect of ROCK2 inhibition on single-cell and collective migratory dynamics. Since PyMT cells were not found to be highly migratory, MDA-MB-231 cells exhibited a significant decrease in single-cell migratory speed upon GV101 treatment. Conversely, with MDA-MB-231 cells lacking a collective migration phenotype, PyMT cells exhibited a subtle reduction in organised cell migration upon ROCK2 inhibition. Interestingly, PyMT cells seeded on matrices previously primed with GV101 showed a more pronounced effect on collective cell migration.
To uncouple the effects of stromal vs epithelial cells during matrix invasion by cancer cells, CAFs were treated with the ROCK2 inhibitor either during the matrix contraction phase (early priming) as opposed to epithelial treatment during the invasion phase (late invasion) or in a continuous manner throughout the contraction and invasion phases. Here, early priming significantly decreased cancer cell invasion at levels comparable to continuous treatment. This suggests that stromal ECM remodelling disruption by GV101 is enough to reduce cancer cell invasion.
ROCK2 expression in triple-negative breast cancers
Analysis of TCGA and METABRIC cohorts reveals a poorer median patient survival with high ROCK2 mRNA expression. Additionally, ROCK2 protein expression is significantly increased in breast cancer progression, indicating that high ROCK2 expression in human triple-negative breast cancer patients correlates with poor patient survival.
Significance of the study:
doi: https://doi.org/10.1242/prelights.42212
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