Immortalization capacity of HPV types is inversely related to chromosomal instability

Abstract

// Denise M. Schütze 1 , Oscar Krijgsman 2 , Peter J.F. Snijders 1 , Bauke Ylstra 1 , Joachim Weischenfeldt 3 , Balca R. Mardin 3 , Adrian M. Stütz 3 , Jan O. Korbel 3 , Johan P. de Winter 4, * , Chris J.L.M. Meijer 1 , Wim G.V. Quint 5 , Leontien Bosch 1 , Saskia M. Wilting 1 , Renske D.M. Steenbergen 1 1 Department of Pathology, VU University Medical Center, Amsterdam, The Netherlands 2 Department of Molecular Oncology, The Netherlands Cancer Institute, Amsterdam, The Netherlands 3 Genome Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany 4 Department of Clinical Genetics, VU University Medical Center, Amsterdam, The Netherlands 5 DDL Diagnostic Laboratory, Voorburg, The Netherlands * JPdeW deceased Correspondence to: Renske D.M. Steenbergen, email: r.steenbergen@vumc.nl Keywords: arrayCGH, high-risk HPV, E6/E7, transformation, chromothripsis Received: November 05, 2015      Accepted: February 28, 2016      Published: March 14, 2016 ABSTRACT High-risk human papillomavirus (hrHPV) types induce immortalization of primary human epithelial cells. Previously we demonstrated that immortalization of human foreskin keratinocytes (HFKs) is HPV type dependent, as reflected by the presence or absence of a crisis period before reaching immortality. This study determined how the immortalization capacity of ten hrHPV types relates to DNA damage induction and overall genomic instability in HFKs. Twenty five cell cultures obtained by transduction of ten hrHPV types (i.e. HPV16/18/31/33/35/45/51/59/66/70 E6E7) in two or three HFK donors each were studied. All hrHPV-transduced HFKs showed an increased number of double strand DNA breaks compared to controls, without exhibiting significant differences between types. However, immortal descendants of HPV-transduced HFKs that underwent a prior crisis period (HPV45/51/59/66/70-transduced HFKs) showed significantly more chromosomal aberrations compared to those without crisis (HPV16/18/31/33/35-transduced HFKs). Notably, the hTERT locus at 5p was exclusively gained in cells with a history of crisis and coincided with increased expression. Chromothripsis was detected in one cell line in which multiple rearrangements within chromosome 8 resulted in a gain of MYC. Together we demonstrated that upon HPV-induced immortalization, the number of chromosomal aberrations is inversely related to the viral immortalization capacity. We propose that hrHPV types with reduced immortalization capacity in vitro , reflected by a crisis period, require more genetic host cell aberrations to facilitate immortalization than types that can immortalize without crisis. This may in part explain the observed differences in HPV-type prevalence in cervical cancers and emphasizes that changes in the host cell genome contribute to HPV-induced carcinogenesis.