Deep sequencing of HPV E6/E7 genes reveals loss of genotypic diversity and gain of clonal dominance in high-grade intraepithelial lesions of the cervix

BMC Genomics

Research Article

Yufeng Wang¹  Graham M. Poage²  Luis Perez²  Jane Shen-Gunther²  Tim H. M. Huang³  Zhao Lai⁴ 

[1] Department of Biology, University of Texas at San Antonio, 78249, San Antonio, TX, USA;Department of Clinical Investigation, Brooke Army Medical Center, Gynecologic Oncology & Clinical Investigation, 3698 Chambers Pass, 78234, Fort Sam Houston, TX, USA;Department of Molecular Medicine, Cancer Therapy and Research Center, University of Texas Health Science Center at San Antonio, 78229, San Antonio, TX, USA;Greehey Children’s Cancer Research Institute, University of Texas Health Science Center at San Antonio, 78229, San Antonio, TX, USA;

关键词: Human papillomavirus;    HPV genotyping;    High-throughput sequencing;    Deep sequencing;    Metagenome;    Virome;    LSIL;    HSIL;   

DOI  :  10.1186/s12864-017-3612-y

 received in 2016-12-19, accepted in 2017-03-07,  发布年份 2017

来源: Springer

【 摘 要 】

BackgroundHuman papillomavirus (HPV) is the carcinogen of almost all invasive cervical cancer and a major cause of oral and other anogenital malignancies. HPV genotyping by dideoxy (Sanger) sequencing is currently the reference method of choice for clinical diagnostics. However, for samples with multiple HPV infections, genotype identification is singular and occasionally imprecise or indeterminable due to overlapping chromatograms. Our aim was to explore and compare HPV metagenomes in abnormal cervical cytology by deep sequencing for correlation with disease states.ResultsLow- and high-grade intraepithelial lesion (LSIL and HSIL) cytology samples were DNA extracted for PCR-amplification of the HPV E6/E7 genes. HPV+ samples were sequenced by dideoxy and deep methods. Deep sequencing revealed ~60% of all samples (n = 72) were multi-HPV infected. Among LSIL samples (n = 43), 27 different genotypes were found. The 3 dominant (most abundant) genotypes were: HPV-39, 11/43 (26%); -16, 9/43 (21%); and -35, 4/43 (9%). Among HSIL (n = 29), 17 HPV genotypes were identified; the 3 dominant genotypes were: HPV-16, 21/29 (72%); -35, 4/29 (14%); and -39, 3/29 (10%). Phylogenetically, type-specific E6/E7 genetic distances correlated with carcinogenic potential. Species diversity analysis between LSIL and HSIL revealed loss of HPV diversity and domination by HPV-16 in HSIL samples.ConclusionsDeep sequencing resolves HPV genotype composition within multi-infected cervical cytology. Biodiversity analysis reveals loss of diversity and gain of dominance by carcinogenic genotypes in high-grade cytology. Metagenomic profiles may therefore serve as a biomarker of disease severity and a population surveillance tool for emerging genotypes.

【 授权许可】

CC BY   
© The Author(s). 2017