Nasopharyngeal carcinoma patient-derived xenograft mouse models reveal potential drugs targeting cell cycle, mTOR, and autophagy pathways

Hsin-Pai Li; Chen-Yang Huang; Kar-Wai Lui; Yin-Kai Chao; Chun-Nan Yeh; Li-Yu Lee; Yenlin Huang; Tung-Liang Lin; Yung-Chia Kuo; Mei-Yuan Huang; Hsien-Chi Fan; An-Chi Lin; Chia-Hsun Hsieh; Kai-Ping Chang; Chien-Yu Lin; Hung-Ming Wang; Mei Chao; Jai-Shin Liu; Yu-Sun Chang; Cheng-Lung Hsu

doi:10.1016/j.tranon.2023.101785

Nasopharyngeal carcinoma patient-derived xenograft mouse models reveal potential drugs targeting cell cycle, mTOR, and autophagy pathways

Transl Oncol. 2023 Dec:38:101785. doi: 10.1016/j.tranon.2023.101785. Epub 2023 Sep 13.

Authors

Hsin-Pai Li¹, Chen-Yang Huang², Kar-Wai Lui³, Yin-Kai Chao⁴, Chun-Nan Yeh⁵, Li-Yu Lee⁶, Yenlin Huang⁶, Tung-Liang Lin², Yung-Chia Kuo², Mei-Yuan Huang⁷, Hsien-Chi Fan², An-Chi Lin², Chia-Hsun Hsieh², Kai-Ping Chang⁸, Chien-Yu Lin⁹, Hung-Ming Wang², Mei Chao¹⁰, Jai-Shin Liu¹¹, Yu-Sun Chang¹², Cheng-Lung Hsu¹³

Affiliations

¹ Department of Microbiology and Immunology, Chang Gung University, Taoyuan 33305, Taiwan; Graduate Institute of Biomedical Sciences, Chang Gung University, Taoyuan 33305, Taiwan; Molecular Medicine Research Center, Chang Gung University, Taoyuan 33305, Taiwan; Division of Hematology-Oncology, Department of Internal Medicine, Chang Gung Memorial Hospital, Chang Gung University, Taoyuan 33305, Taiwan.
² Division of Hematology-Oncology, Department of Internal Medicine, Chang Gung Memorial Hospital, Chang Gung University, Taoyuan 33305, Taiwan.
³ Department of Medical Imaging and Intervention, Chang Gung Memorial Hospital, Chang Gung University, Taoyuan 33305, Taiwan.
⁴ Division of Thoracic and Cardiovascular Surgery, Department of Surgery, Chang Gung Memorial Hospital, Chang Gung University, Taoyuan 33305, Taiwan.
⁵ Department of General Surgery, Liver Research Center, Chang Gung Memorial Hospital, Chang Gung University, Taoyuan 33305, Taiwan.
⁶ Department of Pathology, Chang Gung Memorial Hospital, Chang Gung University, Taoyuan 33305, Taiwan.
⁷ Department of Microbiology and Immunology, Chang Gung University, Taoyuan 33305, Taiwan.
⁸ Department of Otolaryngology-Head and Neck Surgery, Chang Gung Memorial Hospital, Chang Gung University, Taoyuan 33305, Taiwan.
⁹ Department of Radiation, Chang Gung Memorial Hospital, Chang Gung University, Taoyuan 33305, Taiwan.
¹⁰ Department of Microbiology and Immunology, Chang Gung University, Taoyuan 33305, Taiwan; Graduate Institute of Biomedical Sciences, Chang Gung University, Taoyuan 33305, Taiwan; Department of Hepato-Gastroenterology, Chang Gung Memorial Hospital, Chang Gung University, Taoyuan 33305, Taiwan.
¹¹ Department of Biotechnology and Pharmaceutical Technology, Yuanpei University of Medical Technology, Hsinchu City 30015, Taiwan.
¹² Department of Microbiology and Immunology, Chang Gung University, Taoyuan 33305, Taiwan; Molecular Medicine Research Center, Chang Gung University, Taoyuan 33305, Taiwan; Department of Otolaryngology-Head and Neck Surgery, Chang Gung Memorial Hospital, Chang Gung University, Taoyuan 33305, Taiwan.
¹³ Division of Hematology-Oncology, Department of Internal Medicine, Chang Gung Memorial Hospital, Chang Gung University, Taoyuan 33305, Taiwan; School of Medicine, Chang Gung University, Taoyuan 33305, Taiwan. Electronic address: hsu2221@adm.cgmh.org.tw.

Abstract

Background: Nasopharyngeal carcinoma (NPC) is associated with Epstein-Barr virus (EBV) infection. To test preclinical NPC drugs, we established two patient-derived xenograft (PDX) mouse models, EBV-positive PDX-B13 and EBV-negative PDX-Li41, for drug screening.

Methods: Based on next generation sequencing (NGS) studies, PDX-B13 had CCND1 copy number (CN) gain but CDKN2A CN loss, whereas PDX-Li41 had CDKN2A and RB1 CN loss, TSC1 (negative regulator of mTOR) frameshift deletion mutation, and increased activation of mTOR, a serine/threonine kinase that governs metabolism, autophagy, and apoptosis. Increased mTOR was also associated with poor NPC prognosis.

Results: Everolimus, an mTOR inhibitor, suppressed tumor growth in the two PDX NPC models and had an additive antitumor effect with palbociclib, a CDK4/6 inhibitor. PDX tumors treated with various drugs or untreated were subjected to RNA sequencing, transcriptome profile analysis, and selective Western blotting to understand the interactions between these drugs and gene expression profiles. Palbociclib also suppressed EB viral nuclear antigen (EBNA1) expression in PDX-B13. Everolimus together with autophagy inhibitor, hydroxychloroquine, had additive anti-tumor effect on PDX-B13 tumor. Immunohistochemistry revealed that high mTOR levels were correlated with poor overall survival in patients with metastatic NPC (N = 90).

Conclusions: High mTOR levels are a poor prognostic factor in NPC, and cell cycle, mTOR and autophagy pathways may serve as therapeutic targets in NPC. In addition, PDX models can be used for efficiently testing potential NPC drugs.

Keywords: Cell cycle; EBV; NPC; PDX; mTOR.