Establishment and characterization of new tumor xenografts and cancer cell lines from EBV-positive nasopharyngeal carcinoma

Weitao Lin ,Yuk Chun Cheung ,Josephine Mun Yee Ko ,Victor Lee ,Chi Man Tsang ,Wei Dai ,Jaap M Middeldorp ,Pok Man Hau ,Dora Lai-Wan Kwong ,Jun Zhang ,Wen Deng ,Bin Li ,Alan Kwok Shing Chiang ,Chi Keung Chan ,Johnny S H Kwan ,John M Nicholls ,Any Cheung ,Yuan Zhou ,Tengfei Liu ,Honglin Chen ,Yim Ling Yip ,Kwai Fung Hui ,Grace Chung ,Hong Zheng ,Lin Jia ,Jmm Chan ,P Busson ,Sau-Dan Lee ,Kwok Wai Lo ,Kevin Yl Yip ,Siu Tim Cheung ,Xuefeng Li ,Harry Kwok ,Maria Li Lung

doi:10.1038/s41467-018-06889-5

Abstract

The lack of representative nasopharyngeal carcinoma (NPC) models has seriously hampered research on EBV carcinogenesis and preclinical studies in NPC. Here we report the successful growth of five NPC patient-derived xenografts (PDXs) from fifty-eight attempts of transplantation of NPC specimens into NOD/SCID mice. The take rates for primary and recurrent NPC are 4.9% and 17.6%, respectively. Successful establishment of a new EBV-positive NPC cell line, NPC43, is achieved directly from patient NPC tissues by including Rho-associated coiled-coil containing kinases inhibitor (Y-27632) in culture medium. Spontaneous lytic reactivation of EBV can be observed in NPC43 upon withdrawal of Y-27632. Whole-exome sequencing (WES) reveals a close similarity in mutational profiles of these NPC PDXs with their corresponding patient NPC. Whole-genome sequencing (WGS) further delineates the genomic landscape and sequences of EBV genomes in these newly established NPC models, which supports their potential use in future studies of NPC.

Highlights

The lack of representative nasopharyngeal carcinoma (NPC) models has seriously hampered research on Epstein–Barr virus (EBV) carcinogenesis and preclinical studies in NPC
These newly established EBV+ NPC patient-derived xenografts (PDXs) and cell lines significantly recapitulate the mutation profiles of their original NPC tumors, and harbor common genetic alterations reported in NPC, which supports their potential applications in the investigations of NPC pathogenesis
Multiple transfers of NPC xenografts to new mice were usually required before robust growth of the transplanted xenografts could be observed

Summary

Introduction

The lack of representative nasopharyngeal carcinoma (NPC) models has seriously hampered research on EBV carcinogenesis and preclinical studies in NPC. Most if not all the other previously reported NPC cell lines have lost their EBV episomes and became EBV-negative (EBV–ve) upon in vitro propagation[5,6]. We report the successful establishment and comprehensive characterization of four new NPC PDXs (all EBV +ve) and three NPC cell lines (one EBV+ve; two EBV–ve). These newly established EBV+ NPC PDXs and cell lines significantly recapitulate the mutation profiles of their original NPC tumors, and harbor common genetic alterations reported in NPC, which supports their potential applications in the investigations of NPC pathogenesis. Our experience in the establishment of these PDXs and cell lines will facilitate future attempts to generate relevant and representative NPC models for investigations

Methods

Results

Conclusion