Development of Novel Inhibitors of RhoJ to Target Tumour Angiogenesis in Cancer

Abstract

The Rho family of small GTPases orchestrates diverse cellular functions, with members of the Cdc42 subfamily—particularly RhoJ—emerging as pivotal regulators of tumor angiogenesis. Despite its importance, RhoJ remains an underexplored therapeutic target. Through homology modelling of RhoJ and Cdc42 complexed with p21-binding domains (PBDs) from various effectors, combined with extensive (500 ns) molecular dynamics (MD) simulations, a hitherto unreported allosteric pocket in RhoJ is identified in this work, which is absent in homologous members Cdc42 and RhoQ. Virtual screening of FDA-approved drugs against this stable pocket, followed by verification using high-throughput eBRET2 (enhanced bioluminescence resonance energy transfer variant 2) assays, yielded a candidate small-molecule inhibitor with selectivity for RhoJ. Furthermore, RhoJ was predicted to undergo palmitoylation, and MD simulations together with live-cell assays suggest that this lipid modification could regulate its subcellular behaviour and engagement with downstream effectors. Overall, this work uncovers a new therapeutic strategy for targeting tumor angiogenesis via RhoJ inhibition, offering a blueprint for the rational design of anti-angiogenic agents.

Date of Award	9 Dec 2025
Original language	English
Awarding Institution	University of Bristol
Supervisor	Mark P Dodding (Supervisor) & Harry H Mellor (Supervisor)