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Genome-wide profiling of HPV integration in cervical cancer identifies clustered genomic hot spots and a potential microhomology-mediated integration mechanism

Abstract

Human papillomavirus (HPV) integration is a key genetic event in cervical carcinogenesis1. By conducting whole-genome sequencing and high-throughput viral integration detection, we identified 3,667 HPV integration breakpoints in 26 cervical intraepithelial neoplasias, 104 cervical carcinomas and five cell lines. Beyond recalculating frequencies for the previously reported frequent integration sites POU5F1B (9.7%), FHIT (8.7%), KLF12 (7.8%), KLF5 (6.8%), LRP1B (5.8%) and LEPREL1 (4.9%), we discovered new hot spots HMGA2 (7.8%), DLG2 (4.9%) and SEMA3D (4.9%). Protein expression from FHIT and LRP1B was downregulated when HPV integrated in their introns. Protein expression from MYC and HMGA2 was elevated when HPV integrated into flanking regions. Moreover, microhomologous sequence between the human and HPV genomes was significantly enriched near integration breakpoints, indicating that fusion between viral and human DNA may have occurred by microhomology-mediated DNA repair pathways2. Our data provide insights into HPV integration-driven cervical carcinogenesis.

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Figure 1: Distribution of breakpoints in the human and HPV genomes in 135 samples.
Figure 2: Clinical annotation of HPV integration sites in 104 cervical carcinomas, 26 CINs and five cell lines.
Figure 3: Mapping five HPV integration hot spots in 135 samples.
Figure 4: Effects of HPV integration on gene expression in samples assessed by immunohistochemistry.
Figure 5: MH sequences are significantly enriched in the regions flanking integration sites.

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Sequence Read Archive

Referenced accessions

NCBI Reference Sequence

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Acknowledgements

This work was supported by funds from the National Development Program (973) for the Key Basic Research of China (2015CB553903 and 2013CB911304) and the National Natural Science Funding of China (81230038, 81372805, 81172466, 81272859, 81372801, 81372804, 81402158, 81370469, 81372806, 81370469, 81172468, 81101964, 81230052 and 81302266). The study is also sponsored by the Chinese 863 Program (2012AA02A507, 2012AA02A201), Guangdong Enterprise Key Laboratory of Human Disease Genomics, ShenZhen Engineering Laboratory for Clinical Molecular Diagnostic and China National GeneBank–Shenzhen. We thank all participants recruited for this study. We thank our colleagues from BGI, H. Huang, L. Huang, X. Zhuang, L. Lin, H. Cao, X. Fang, X. Zhang, Y. Shuang and H. Yang for sequencing and analysis.

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D.M. took full responsibility for the study, especially in conceiving, designing and supervising the research together with Z.H. and H.W. W.W., J.Z., J.W. and X.X. participated in the study design. G.C., Q.G., Shuang Li., L.X., C.W., S. Liao, X.M., P.W., K.L. and S.W. supervised the diagnosis of patients and subject recruitment. D.Z., H.S., W.J., W.L., X.Z., H.L., X.F., Shuaicheng Li and X.L. performed statistical analysis. W.L., X.Z. and Y.Z. performed HIVID and RNA-seq analysis. W.J. performed FuseSV (in-house software for MH analysis) and WGS analysis. W.D., L.Y., L.W., H.S., X.W., C.Z., W.C., T.T., A.F. and Z.W. performed the experiments. The manuscript was drafted by Z.H., D.Z., W.J., W.L. and X.Z. under the supervision of D.M., H.W. and X.X. All authors critically reviewed the article and approved the final manuscript.

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Correspondence to Xun Xu or Hui Wang or Ding Ma.

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The authors declare no competing financial interests.

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Hu, Z., Zhu, D., Wang, W. et al. Genome-wide profiling of HPV integration in cervical cancer identifies clustered genomic hot spots and a potential microhomology-mediated integration mechanism. Nat Genet 47, 158–163 (2015). https://doi.org/10.1038/ng.3178

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