Combining peptide TNIIIA2 with all- trans retinoic acid accelerates N-Myc protein degradation and neuronal differentiation in MYCN-amplified neuroblastoma cells

Kazuki Otsuka; Manabu Sasada; Takuya Iyoda; Yusuke Nohara; Shunsuke Sakai; Tatsufumi Asayama; Yusuke Suenaga; Sana Yokoi; Yoshikazu Higami; Hiroaki Kodama; Fumio Fukai

Combining peptide TNIIIA2 with all- trans retinoic acid accelerates N-Myc protein degradation and neuronal differentiation in MYCN-amplified neuroblastoma cells

Am J Cancer Res. 2019 Feb 1;9(2):434-448. eCollection 2019.

Authors

Kazuki Otsuka¹, Manabu Sasada^{1

2

3}, Takuya Iyoda⁴, Yusuke Nohara¹, Shunsuke Sakai¹, Tatsufumi Asayama¹, Yusuke Suenaga³, Sana Yokoi³, Yoshikazu Higami^{2

5}, Hiroaki Kodama⁶, Fumio Fukai^{1

2}

Affiliations

¹ Department of Molecular Patho-Physiology, Faculty of Pharmaceutical Sciences, Tokyo University of Science Noda, Chiba, Japan.
² Translational Research Center, Research Institutes for Science and Technology, Tokyo University of Science Noda, Chiba, Japan.
³ Cancer Genome Center, Chiba Cancer Center Research Institute Chiba, Japan.
⁴ Department of Pharmacy, Faculty of Pharmaceutical Sciences, Sanyo-Onoda City University Sanyo-Onoda, Yamaguchi, Japan.
⁵ Department of Molecular Pathology and Metabolic Disease, Faculty of Pharmaceutical Sciences, Tokyo University of Science Noda, Chiba, Japan.
⁶ Department of Biochemistry, Faculty of Science and Engineering, Saga University Saga, Japan.

PMID: 30906641
PMCID: PMC6405964

Abstract

Neuroblastoma is one of the common solid tumors of childhood. Nearly half of neuroblastoma patients are classified into the high-risk group, and their 5-year event-free survival (EFS) rates remain unsatisfactory in the range of 30-40%. High-risk neuroblastoma is characterized by amplification of the MYCN gene and excessive expression of its protein product, N-Myc. Because N-Myc is a transcription factor for various pro-proliferative proteins, the excessive expression causes aberrant or blocked neuronal differentiation during development of sympathetic nervous system, which is a central aspect of neuroblastoma genesis. The current main treatment for high-risk neuroblastoma is intensive chemotherapy using anti-cancer drugs that induce apoptosis in tumor cells, but intensive chemotherapy has another serious risk of long-lasting side effects, so-called "late effects", that occur many years after chemotherapy has ended. As a solution for such situation, differentiation therapy has been expected as a mild chemotherapy with a low risk of late effects, and an application of retinoic acid (RA) and its derivatives as treatment for high-risk neuroblastoma has long been attempted. However, the clinical outcome has not been sufficient with the use of retinoids, including all-trans retinoic acid (ATRA), mainly because of the inhibition of differentiation caused by N-Myc. In the present study, we succeeded in synergistically accelerating the ATRA-induced neuronal differentiation of MYCN-amplified neuroblastoma cells by combining a peptide derived from tenascin-C, termed TNIIIA2, which has a potent ability to activate β1-integrins. Accelerated differentiation was caused by a decrease in N-Myc protein level in neuroblastoma cells after the combined treatment of TNIIIA2 with ATRA. That is, combination treatment using ATRA with TNIIIA2 induced proteasomal degradation in the N-Myc oncoprotein of neuroblastoma cells with MYCN gene amplification, and this caused acceleration of neuronal differentiation and attenuation of malignant properties. Furthermore, an in vivo experiment using a xenograft mouse model showed a therapeutic potential of the combination administration of ATRA and TNIIIA2 for high-risk neuroblastoma. These results provide a new insight into differentiation therapy for high-risk neuroblastoma based on N-Myc protein degradation.

Keywords: ATRA; N-Myc; Neuroblastoma; differentiation therapy; integrin; mycn; proto-oncogene; retinoic acid; tenascin-C; ubiquitin-proteasome system.