The ROCK-1/2 inhibitor RKI-1447 blocks N-MYC, promotes cell death, and emerges as a synergistic partner for BET inhibitors in neuroblastoma

Cancer Lett. 2024 Nov 28:605:217261. doi: 10.1016/j.canlet.2024.217261. Epub 2024 Sep 20.

Abstract

High-risk neuroblastoma has a poor prognosis despite intensive treatment, highlighting the need for new therapeutic strategies. Genetic alterations in activators and inactivators of Rho GTPase have been identified in neuroblastoma suggested to activate Rho/Rho-kinase (ROCK) signaling. ROCK has also been implicated in therapy resistance. Therefore, we have explored the efficacy of the dual ROCK inhibitor RKI-1447 in neuroblastoma, emphasizing combination strategies. Treatment with RKI-1447 resulted in decreased growth, increased cell death, and inhibition of N-MYC in vitro and in vivo. A combination screen revealed enhanced effects between RKI-1447 and BET inhibitors. Synergistic effects from RKI-1447 and the BET inhibitor, ABBV-075, were confirmed in various neuroblastoma models, including zebrafish. Interestingly, ABBV-075 increased phosphorylation of both myosin light chain 2 and cofilin, downstream effectors of ROCK, increases that were blocked by adding RKI-1447. The combination treatment also augmented an inhibitory effect on C-MYC and, less pronounced, N-MYC protein expression. BET inhibitors have shown preclinical efficacy against neuroblastoma, but acquired resistance has limited their therapeutic benefit. We reveal that the combination of ROCK and BET inhibitors offers a promising treatment approach that can potentially mitigate resistance to BET inhibitors and reduce toxicity.

Keywords: BET inhibitor; Drug combinations; Neuroblastoma; RKI-1447; ROCK inhibitor.

MeSH terms

  • Actin Depolymerizing Factors / metabolism
  • Animals
  • Antineoplastic Combined Chemotherapy Protocols / pharmacology
  • Apoptosis / drug effects
  • Bromodomain Containing Proteins
  • Cardiac Myosins
  • Cell Death / drug effects
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Drug Synergism*
  • Humans
  • Mice
  • Myosin Light Chains / metabolism
  • N-Myc Proto-Oncogene Protein / genetics
  • N-Myc Proto-Oncogene Protein / metabolism
  • Neuroblastoma* / drug therapy
  • Neuroblastoma* / genetics
  • Neuroblastoma* / metabolism
  • Neuroblastoma* / pathology
  • Phosphorylation / drug effects
  • Protein Kinase Inhibitors / pharmacology
  • Proteins
  • Proto-Oncogene Proteins c-myc / genetics
  • Proto-Oncogene Proteins c-myc / metabolism
  • Signal Transduction / drug effects
  • Xenograft Model Antitumor Assays
  • Zebrafish*
  • rho-Associated Kinases* / antagonists & inhibitors
  • rho-Associated Kinases* / metabolism

Substances

  • rho-Associated Kinases
  • ROCK2 protein, human
  • ROCK1 protein, human
  • N-Myc Proto-Oncogene Protein
  • Protein Kinase Inhibitors
  • myosin light chain 2
  • Myosin Light Chains
  • MYCN protein, human
  • bromodomain and extra-terminal domain protein, human
  • Actin Depolymerizing Factors
  • Proto-Oncogene Proteins c-myc
  • Bromodomain Containing Proteins
  • Proteins
  • Cardiac Myosins