Splicing Modulation Results in Aberrant Isoforms and Protein Products of p53 Pathway Genes and the Sensitization of B Cells to Non-Genotoxic MDM2 Inhibition

Int J Mol Sci. 2023 Jan 26;24(3):2410. doi: 10.3390/ijms24032410.

Abstract

Several molecular subtypes of cancer are highly dependent on splicing for cell survival. There is a general interest in the therapeutic targeting of splicing by small molecules. E7107, a first-in-class spliceosome inhibitor, showed strong growth inhibitory activities against a large variety of human cancer xenografts. Chronic lymphocytic leukaemia (CLL) is a clinically heterogeneous hematologic malignancy, with approximately 90% of cases being TP53 wild-type at diagnosis. An increasing number of studies are evaluating alternative targeted agents in CLL, including MDM2-p53 binding antagonists. In this study, we report the effect of splicing modulation on key proteins in the p53 signalling pathway, an important cell death pathway in B cells. Splicing modulation by E7107 treatment reduced full-length MDM2 production due to exon skipping, generating a consequent reciprocal p53 increase in TP53WT cells. It was especially noteworthy that a novel p21WAF1 isoform with compromised cyclin-dependent kinase inhibitory activity was produced due to intron retention. E7107 synergized with the MDM2 inhibitor RG7388, via dual MDM2 inhibition; by E7107 at the transcript level and by RG7388 at the protein level, producing greater p53 stabilisation and apoptosis. This study provides evidence for a synergistic MDM2 and spliceosome inhibitor combination as a novel approach to treat CLL and potentially other haematological malignancies.

Keywords: E7107; MDM2–p53 antagonists; RG7388 (idasanutlin); p53 pathway; splicing modulation.

MeSH terms

  • Antineoplastic Agents* / pharmacology
  • Apoptosis / genetics
  • B-Lymphocytes* / metabolism
  • Humans
  • Leukemia, Lymphocytic, Chronic, B-Cell* / drug therapy
  • Leukemia, Lymphocytic, Chronic, B-Cell* / genetics
  • Protein Isoforms / metabolism
  • Proto-Oncogene Proteins c-mdm2 / metabolism
  • Pyrrolidines / pharmacology
  • Tumor Suppressor Protein p53 / metabolism

Substances

  • Antineoplastic Agents
  • MDM2 protein, human
  • Protein Isoforms
  • Proto-Oncogene Proteins c-mdm2
  • Pyrrolidines
  • RG7388
  • Tumor Suppressor Protein p53

Grants and funding

This study was supported by Bloodwise (grant # 13034), the JGW Patterson Foundation (grant # BH152495) and the Newcastle Healthcare Charity (grant # BH152694). The Turkish Ministry of National Education funded the first author’s Ph.D. study.