Targeting the TP53/MDM2 axis enhances radiation sensitivity in atypical teratoid rhabdoid tumors

Alimova,Irina; Wang,Dong; Danis,Etienne; Pierce,Angela; Donson,Andrew; Serkova,Natalie; Madhavan,Krishna; Lakshmanachetty,Senthilnath; Balakrishnan,Ilango; Foreman,Nicholas K.; Mitra,Siddhartha; Venkataraman,Sujatha; Vibhakar,Rajeev

doi:10.3892/ijo.2022.5322

Targeting the TP53/MDM2 axis enhances radiation sensitivity in atypical teratoid rhabdoid tumors

Authors:
- Irina Alimova
- Dong Wang
- Etienne Danis
- Angela Pierce
- Andrew Donson
- Natalie Serkova
- Krishna Madhavan
- Senthilnath Lakshmanachetty
- Ilango Balakrishnan
- Nicholas K. Foreman
- Siddhartha Mitra
- Sujatha Venkataraman
- Rajeev Vibhakar
View Affiliations

Affiliations: Department of Pediatrics, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA, Department of Radiology, University of Colorado School of Medicine, Aurora, CO 80045, USA
Published online on: February 17, 2022 https://doi.org/10.3892/ijo.2022.5322
Article Number: 32
Copyright: © Alimova et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Atypical teratoid rhabdoid tumor (ATRT) is a highly aggressive pediatric brain tumor. Despite radiation, aggressive chemotherapy and autologous stem cell rescue, children usually have a poor survival time. In the present study, the role of TP53/MDM2 interaction in ATRT was investigated. A functional genomic screen identified the TP53/MDM2 axis as a therapeutic target in the central nervous system (CNS) ATRT. Gene expression analysis revealed that all ATRT sub‑groups expressed high levels of MDM2, which is a negative regulator of TP53. Using cell viability, colony formation and methylcellulose assays it was found that genetic MDM2 inhibition with short hairpin RNA or chemical MDM2 inhibition with small molecule inhibitors, Nutlin3 and idasanutlin (RG7388) decreased the growth of ATRT cell lines. Furthermore, idasanutlin significantly decreased the growth of intracranial orthotopic ATRT brain tumors, as evaluated using T2 MRI, and prolonged survival time relative to control animals. MRI of intracranial tumors showed that diffusion coefficient, an effective marker for successful treatment, significantly increased with idasanutlin treatment showing tumor necrosis/apoptosis. Immunohistochemistry revealed an increased number of caspase‑3‑positive cells in the idasanutlin treatment group, confirming the induction of apoptosis in vivo. Using flow cytometry and western blot analysis we show that inhibition of MDM2 enhanced radiation sensitivity in vitro by potentiating DNA damage via the induction of the TP53/Bax/Puma proapoptotic axis. Furthermore, DNA damage was associated with increased mitochondrial reactive oxygen species accumulation. The present study demonstrated that MDM2 expression level was increased in ATRT patient samples and MDM2 inhibition suppressed ATRT cell growth in vitro, and leads to apoptosis in vivo. MDM2 inhibition potentiates DNA damage and sensitizes ATRT cells to radiation. These findings highlight the TP53/MDM2 axis as a rational therapeutic target in CNS ATRT.

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