A novel combination therapy targeting ubiquitin-specific protease 5 in MYCN-driven neuroblastoma

• Published in: Oncogene 2021-04, Vol.40 (13), p.2367-2381
• Main Authors: Cheung, Belamy B., Kleynhans, Ane, Mittra, Rituparna, Kim, Patrick Y., Holien, Jessica K., Nagy, Zsuzsanna, Ciampa, Olivia C., Seneviratne, Janith A., Mayoh, Chelsea, Raipuria, Mukesh, Gadde, Satyanarayana, Massudi, Hassina, Wong, Iris Poh Ling, Tan, Owen, Gong, Andrew, Suryano, Aldwin, Diakiw, Sonya M., Liu, Bing, Arndt, Greg M., Liu, Tao, Kumar, Naresh, Sangfelt, Olle, Zhu, Shizhen, Norris, Murray D., Haber, Michelle, Carter, Daniel R., Parker, Michael W., Marshall, Glenn M.
• Format: Article
• Language: English
• Subjects: 101/58; 13/1; 13/2; 38/47; 42/109; 42/89; 45/77; 631/154/1435/2163; 631/154/555; 631/67/2332; 64/116; 64/60; 82/80; 96/106; Animals; Animals, Genetically Modified - genetics; Apoptosis; Apoptosis - drug effects; Cancer; Carcinogenesis - drug effects; Cell Biology; Cell Proliferation - drug effects; Cell Survival - drug effects; Cell viability; Combination therapy; Cytotoxicity; Drug development; Drug Evaluation, Preclinical; Drug screening; Drug therapy; Drug therapy, Combination; Gene expression; Gene Expression Regulation, Neoplastic - drug effects; Genetic aspects; Health aspects; Heterografts; Histone deacetylase; Histone Deacetylase Inhibitors - pharmacology; Human Genetics; Humans; Hydroxamic acid; Internal Medicine; Medicine; Medicine & Public Health; Methods; Mice; Molecular targeted therapy; N-Myc Proto-Oncogene Protein - genetics; Neuroblastoma; Neuroblastoma - drug therapy; Neuroblastoma - genetics; Neuroblastoma - pathology; Neuroblasts; Oncogenes; Oncology; Oncology, Experimental; Proteases; Small Molecule Libraries - pharmacology; Therapeutic targets; Transgenic mice; Tumorigenesis; Ubiquitin; Ubiquitin-Specific Proteases - genetics; Ubiquitin-specific proteinase; Ubiquitination; Vorinostat - pharmacology; Xenografts; Zebrafish - genetics; Zebrafish Proteins - genetics
• ISSN: 0950-9232; 1476-5594
• DOI: 10.1038/s41388-021-01712-w

Histone deacetylase (HDAC) inhibitors are effective in MYCN-driven cancers, because of a unique need for HDAC recruitment by the MYCN oncogenic signal. However, HDAC inhibitors are much more effective in combination with other anti-cancer agents. To identify novel compounds which act synergistically with HDAC inhibitor, such as suberanoyl hydroxamic acid (SAHA), we performed a cell-based, high-throughput drug screen of 10,560 small molecule compounds from a drug-like diversity library and identified a small molecule compound (SE486-11) which synergistically enhanced the cytotoxic effects of SAHA. Effects of drug combinations on cell viability, proliferation, apoptosis and colony forming were assessed in a panel of neuroblastoma cell lines. Treatment with SAHA and SE486-11 increased MYCN ubiquitination and degradation, and markedly inhibited tumorigenesis in neuroblastoma xenografts, and, MYCN transgenic zebrafish and mice. The combination reduced ubiquitin-specific protease 5 (USP5) levels and increased unanchored polyubiquitin chains. Overexpression of USP5 rescued neuroblastoma cells from the cytopathic effects of the combination and reduced unanchored polyubiquitin, suggesting USP5 is a therapeutic target of the combination. SAHA and SE486-11 directly bound to USP5 and the drug combination exhibited a 100-fold higher binding to USP5 than individual drugs alone in microscale thermophoresis assays. MYCN bound to the USP5 promoter and induced USP5 gene expression suggesting that USP5 and MYCN expression created a forward positive feedback loop in neuroblastoma cells. Thus, USP5 acts as an oncogenic cofactor with MYCN in neuroblastoma and the novel combination of HDAC inhibitor with SE486-11 represents a novel therapeutic approach for the treatment of MYCN-driven neuroblastoma.