The MDM2 inhibitor CGM097 combined with the BET inhibitor OTX015 induces cell death and inhibits tumor growth in models of neuroblastoma

Cancer Med. 2020 Nov;9(21):8144-8158. doi: 10.1002/cam4.3407. Epub 2020 Oct 9.

Abstract

Background: Neuroblastoma (NB) is the most common extracranial solid tumor in infants and children, with amplification of the oncogene MYCN being a hallmark of high-risk disease and poor prognosis. Although less frequent, overexpression of MYC is similarly an indicator of poor prognosis. Most NB tumors initially respond to chemotherapy, however, most will relapse, resulting in chemoresistant disease. After relapse, there is growing evidence of p53 inactivation. MYC/MYCN and MDM2 have been shown to interact and contribute to NB growth and disease progression. MDM2 inhibitors and Bromodomain and Extra-Terminal domain (BET) inhibitors have both shown promise in treating NB by increasing the expression of p53 and decreasing MYC/MYCN expression, respectively. Our study focuses on the combined treatment of a MDM2 inhibitor (CGM097) with a BET inhibitor (OTX015) in neuroblastoma.

Methods: Two p53 wild-type and two p53 mutant established neuroblastoma cells lines were used to test this combination. Ray design assays were used to test whether this combination was synergistically cytotoxic to NB cells. Western blots were performed to check signaling pathways of interest after drug treatment. IncuCyte imaging and flow cytometry were utilized to quantify the apoptotic and cytostatic effects of these drugs on NB cells. In vivo studies were carried out to test the antitumor effect of this combination in a living host.

Results: The combination of CGM097 and OTX015 resulted in p53 activation, decreased expression of MYC family proteins and a subsequent synergistic increase in NB cell death.

Conclusion: This study warrants further investigation into the combination of MDM2 inhibitors and BET inhibitors for the treatment in NB.

Keywords: BET; MDM2; MYC; MYCN; neuroblastoma; p53.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetanilides / administration & dosage
  • Acetanilides / pharmacology*
  • Animals
  • Antineoplastic Combined Chemotherapy Protocols
  • Apoptosis / drug effects
  • Caspase 3 / metabolism
  • Caspase 7 / metabolism
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Drug Synergism
  • Enzyme Activation / drug effects
  • Female
  • G1 Phase Cell Cycle Checkpoints / drug effects
  • Heterocyclic Compounds, 3-Ring / administration & dosage
  • Heterocyclic Compounds, 3-Ring / pharmacology*
  • Humans
  • Isoquinolines / administration & dosage
  • Isoquinolines / pharmacology*
  • Mice
  • Neoplasm Transplantation
  • Neuroblastoma / drug therapy*
  • Piperazines / administration & dosage
  • Piperazines / pharmacology*
  • Proteins / antagonists & inhibitors*
  • Proteins / metabolism
  • Proto-Oncogene Proteins c-mdm2 / antagonists & inhibitors*
  • Proto-Oncogene Proteins c-mdm2 / metabolism
  • Proto-Oncogene Proteins c-myc / metabolism
  • Signal Transduction
  • Tumor Suppressor Protein p53 / metabolism

Substances

  • Acetanilides
  • Heterocyclic Compounds, 3-Ring
  • Isoquinolines
  • NVP-CGM097
  • OTX015
  • Piperazines
  • Proteins
  • Proto-Oncogene Proteins c-myc
  • Tumor Suppressor Protein p53
  • bromodomain and extra-terminal domain protein, human
  • MDM2 protein, human
  • Proto-Oncogene Proteins c-mdm2
  • Caspase 3
  • Caspase 7