HPV infection alters vaginal microbiome through down-regulating host mucosal innate peptides used by Lactobacilli as amino acid sources.

Amino Acids Animals Female Follow-Up Studies Lactobacillus / physiology Mice Microbiota / physiology Mucous Membrane Papillomavirus Infections Peptides Retrospective Studies Vagina / microbiology Vaginosis, Bacterial / microbiology

Journal

Nature communications

ISSN: 2041-1723

Titre abrégé: Nat Commun

Pays: England

ID NLM: 101528555

Informations de publication

Date de publication:
28 02 2022

Historique:

received: 10 04 2020

accepted: 03 02 2022

entrez: 1 3 2022

pubmed: 2 3 2022

medline: 13 4 2022

Statut: epublish

Résumé

Despite the high prevalence of both cervico-vaginal human papillomavirus (HPV) infection and bacterial vaginosis (BV) worldwide, their causal relationship remains unclear. While BV has been presumed to be a risk factor for HPV acquisition and related carcinogenesis for a long time, here, supported by both a large retrospective follow-up study (n = 6,085) and extensive in vivo data using the K14-HPV16 transgenic mouse model, we report a novel blueprint in which the opposite association also exists. Mechanistically, by interacting with several core members (NEMO, CK1 and β-TrCP) of both NF-κB and Wnt/β-catenin signaling pathways, we show that HPV E7 oncoprotein greatly inhibits host defense peptide expression. Physiologically secreted by the squamous mucosa lining the lower female genital tract, we demonstrate that some of these latter are fundamental factors governing host-microbial interactions. More specifically, several innate molecules down-regulated in case of HPV infection are hydrolyzed, internalized and used by the predominant Lactobacillus species as amino acid source sustaining their growth/survival. Collectively, this study reveals a new viral immune evasion strategy which, by its persistent/negative impact on lactic acid bacteria, ultimately causes the dysbiosis of vaginal microbiota.

Identifiants

DOI: 10.1038/s41467-022-28724-8 PMID: 35228537 PMC: PMC8885657

pubmed: 35228537

doi: 10.1038/s41467-022-28724-8

pii: 10.1038/s41467-022-28724-8

pmc: PMC8885657

doi:

Substances chimiques

Amino Acids 0

Peptides 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

Pagination

1076

Informations de copyright

Références

de Sanjose, S. et al. Worldwide prevalence and genotype distribution of cervical human papillomavirus DNA in women with normal cytology: a meta-analysis. Lancet Infect. Dis. 7, 453–459 (2007).

pubmed: 17597569 doi: 10.1016/S1473-3099(07)70158-5

de Martel, C., Plummer, M., Vignat, J. & Franceschi, S. Worldwide burden of cancer attributable to HPV by site, country and HPV type. International journal of cancer. J. Int. du cancer 141, 664–670 (2017).

doi: 10.1002/ijc.30716

Schiffman, M. et al. Carcinogenic human papillomavirus infection. Nat. Rev. Dis. Prim. 2, 16086 (2016).

pubmed: 27905473 doi: 10.1038/nrdp.2016.86

Luo, X. et al. HPV16 drives cancer immune escape via NLRX1-mediated degradation of STING. J. Clin. Investig. 130, 1635–1652 (2020).

pubmed: 31874109 pmcid: 7108911 doi: 10.1172/JCI129497

Chang, Y. E. & Laimins, L. A. Microarray analysis identifies interferon-inducible genes and Stat-1 as major transcriptional targets of human papillomavirus type 31. J. Virol. 74, 4174–4182 (2000).

pubmed: 10756030 pmcid: 111932 doi: 10.1128/JVI.74.9.4174-4182.2000

Reiser, J. et al. High-risk human papillomaviruses repress constitutive kappa interferon transcription via E6 to prevent pathogen recognition receptor and antiviral-gene expression. J. Virol. 85, 11372–11380 (2011).

pubmed: 21849431 pmcid: 3194958 doi: 10.1128/JVI.05279-11

Ronco, L. V., Karpova, A. Y., Vidal, M. & Howley, P. M. Human papillomavirus 16 E6 oncoprotein binds to interferon regulatory factor-3 and inhibits its transcriptional activity. Genes Dev. 12, 2061–2072 (1998).

pubmed: 9649509 pmcid: 316980 doi: 10.1101/gad.12.13.2061

Bottley, G. et al. High-risk human papillomavirus E7 expression reduces cell-surface MHC class I molecules and increases susceptibility to natural killer cells. Oncogene 27, 1794–1799 (2008).

pubmed: 17828295 doi: 10.1038/sj.onc.1210798

Hasan, U. A. et al. TLR9 expression and function is abolished by the cervical cancer-associated human papillomavirus type 16. J. Immunol. 178, 3186–3197 (2007).

pubmed: 17312167 doi: 10.4049/jimmunol.178.5.3186

Karim, R. et al. Human papillomavirus deregulates the response of a cellular network comprising of chemotactic and proinflammatory genes. PLoS ONE 6, e17848 (2011).

pubmed: 21423754 pmcid: 3056770 doi: 10.1371/journal.pone.0017848

Human Microbiome Project, C. Structure, function and diversity of the healthy human microbiome. Nature 486, 207–214 (2012).

doi: 10.1038/nature11234

Zhou, X. et al. Characterization of vaginal microbial communities in adult healthy women using cultivation-independent methods. Microbiology 150, 2565–2573 (2004).

pubmed: 15289553 doi: 10.1099/mic.0.26905-0

Anahtar, M. N., Gootenberg, D. B., Mitchell, C. M. & Kwon, D. S. Cervicovaginal microbiota and reproductive health: the virtue of simplicity. Cell host microbe 23, 159–168 (2018).

pubmed: 29447695 doi: 10.1016/j.chom.2018.01.013

Onderdonk, A. B., Delaney, M. L. & Fichorova, R. N. The human microbiome during bacterial vaginosis. Clin. Microbiol. Rev. 29, 223–238 (2016).

pubmed: 26864580 pmcid: 4786887 doi: 10.1128/CMR.00075-15

Turovskiy, Y., Sutyak Noll, K. & Chikindas, M. L. The aetiology of bacterial vaginosis. J. Appl. Microbiol. 110, 1105–1128 (2011).

pubmed: 21332897 pmcid: 3072448 doi: 10.1111/j.1365-2672.2011.04977.x

Muzny, C. A. & Schwebke, J. R. Pathogenesis of bacterial vaginosis: discussion of current hypotheses. J. Infect. Dis. 214, S1–S5 (2016).

pubmed: 27449868 pmcid: 4957507 doi: 10.1093/infdis/jiw121

Leitich, H. et al. Bacterial vaginosis as a risk factor for preterm delivery: a meta-analysis. Am. J. Obstet. Gynecol. 189, 139–147 (2003).

pubmed: 12861153 doi: 10.1067/mob.2003.339

Sweet, R. L. Gynecologic conditions and bacterial vaginosis: implications for the non-pregnant patient. Infect. Dis. Obstet. Gynecol. 8, 184–190 (2000).

pubmed: 10968604 pmcid: 1784684 doi: 10.1155/S1064744900000260

Cherpes, T. L., Meyn, L. A., Krohn, M. A., Lurie, J. G. & Hillier, S. L. Association between acquisition of herpes simplex virus type 2 in women and bacterial vaginosis. Clin. Infect. Dis.: Off. Publ. Infect. Dis. Soc. Am. 37, 319–325 (2003).

doi: 10.1086/375819

Cohen, C. R. et al. Bacterial vaginosis associated with increased risk of female-to-male HIV-1 transmission: a prospective cohort analysis among African couples. PLoS Med. 9, e1001251 (2012).

pubmed: 22745608 pmcid: 3383741 doi: 10.1371/journal.pmed.1001251

Wiesenfeld, H. C., Hillier, S. L., Krohn, M. A., Landers, D. V. & Sweet, R. L. Bacterial vaginosis is a strong predictor of Neisseria gonorrhoeae and Chlamydia trachomatis infection. Clin. Infect. Dis.: Off. Publ. Infect. Dis. Soc. Am. 36, 663–668 (2003).

doi: 10.1086/367658

Brotman, R. M. et al. Interplay between the temporal dynamics of the vaginal microbiota and human papillomavirus detection. J. Infect. Dis. 210, 1723–1733 (2014).

pubmed: 24943724 pmcid: 4296189 doi: 10.1093/infdis/jiu330

Briselden, A. M., Moncla, B. J., Stevens, C. E. & Hillier, S. L. Sialidases (neuraminidases) in bacterial vaginosis and bacterial vaginosis-associated microflora. J. Clin. Microbiol. 30, 663–666 (1992).

pubmed: 1551983 pmcid: 265128 doi: 10.1128/jcm.30.3.663-666.1992

Mitra, A. et al. The vaginal microbiota associates with the regression of untreated cervical intraepithelial neoplasia 2 lesions. Nat. Commun. 11, 1999 (2020).

pubmed: 32332850 pmcid: 7181700 doi: 10.1038/s41467-020-15856-y

Brusselaers, N., Shrestha, S., van de Wijgert, J. & Verstraelen, H. Vaginal dysbiosis and the risk of human papillomavirus and cervical cancer: systematic review and meta-analysis. Am. J. Obstet. Gynecol. 221, 9–18 e18 (2019).

pubmed: 30550767 doi: 10.1016/j.ajog.2018.12.011

Gillet, E. et al. Bacterial vaginosis is associated with uterine cervical human papillomavirus infection: a meta-analysis. BMC Infect. Dis. 11, 10 (2011).

pubmed: 21223574 pmcid: 3023697 doi: 10.1186/1471-2334-11-10

Kyrgiou, M., Mitra, A. & Moscicki, A. B. Does the vaginal microbiota play a role in the development of cervical cancer? Transl. Res.: J. Lab. Clin. Med. 179, 168–182 (2017).

doi: 10.1016/j.trsl.2016.07.004

Liang, Y., Chen, M., Qin, L., Wan, B. & Wang, H. A meta-analysis of the relationship between vaginal microecology, human papillomavirus infection and cervical intraepithelial neoplasia. Infect. agents cancer 14, 29 (2019).

doi: 10.1186/s13027-019-0243-8

Yarbrough, V. L., Winkle, S. & Herbst-Kralovetz, M. M. Antimicrobial peptides in the female reproductive tract: a critical component of the mucosal immune barrier with physiological and clinical implications. Hum. Reprod. Update 21, 353–377 (2015).

pubmed: 25547201 doi: 10.1093/humupd/dmu065

Wang, G., Li, X. & Wang, Z. APD3: the antimicrobial peptide database as a tool for research and education. Nucleic Acids Res. 44, D1087–D1093 (2016).

pubmed: 26602694 doi: 10.1093/nar/gkv1278

Tsutsumi-Ishii, Y. & Nagaoka, I. NF-kappa B-mediated transcriptional regulation of human beta-defensin-2 gene following lipopolysaccharide stimulation. J. Leukoc. Biol. 71, 154–162 (2002).

pubmed: 11781391 doi: 10.1189/jlb.71.1.154

Johansen, C., Bertelsen, T., Ljungberg, C., Mose, M. & Iversen, L. Characterization of TNF-alpha- and IL-17A-mediated synergistic induction of DEFB4 gene expression in human keratinocytes through IkappaBzeta. J. Investig. Dermatol. 136, 1608–1616 (2016).

pubmed: 27117051 doi: 10.1016/j.jid.2016.04.012

Clauss, A. et al. Overexpression of elafin in ovarian carcinoma is driven by genomic gains and activation of the nuclear factor kappaB pathway and is associated with poor overall survival. Neoplasia 12, 161–172 (2010).

pubmed: 20126474 pmcid: 2814354 doi: 10.1593/neo.91542

Dreos, R., Ambrosini, G., Perier, R. C. & Bucher, P. The Eukaryotic Promoter Database: expansion of EPDnew and new promoter analysis tools. Nucleic acids Res. 43, D92–D96 (2015).

pubmed: 25378343 doi: 10.1093/nar/gku1111

Dang, C. V. et al. The c-Myc target gene network. Semin. cancer Biol. 16, 253–264 (2006).

pubmed: 16904903 doi: 10.1016/j.semcancer.2006.07.014

Fernandez, P. C. et al. Genomic targets of the human c-Myc protein. Genes Dev. 17, 1115–1129 (2003).

pubmed: 12695333 pmcid: 196049 doi: 10.1101/gad.1067003

He, T. C. et al. Identification of c-MYC as a target of the APC pathway. Science 281, 1509–1512 (1998).

pubmed: 9727977 doi: 10.1126/science.281.5382.1509

Jeanes, A., Gottardi, C. J. & Yap, A. S. Cadherins and cancer: how does cadherin dysfunction promote tumor progression? Oncogene 27, 6920–6929 (2008).

pubmed: 19029934 pmcid: 2745643 doi: 10.1038/onc.2008.343

Schroeder, B. O. et al. Paneth cell alpha-defensin 6 (HD-6) is an antimicrobial peptide. Mucosal Immunol. 8, 661–671 (2015).

pubmed: 25354318 doi: 10.1038/mi.2014.100

Schroeder, B. O. et al. Reduction of disulphide bonds unmasks potent antimicrobial activity of human beta-defensin 1. Nature 469, 419–423 (2011).

pubmed: 21248850 doi: 10.1038/nature09674

Torcia, M. G. Interplay among vaginal microbiome, immune response and sexually transmitted viral infections. Int. J. Mol. Sci. 20, (2019).

Lu, H. et al. Characteristics of bacterial vaginosis infection in cervical lesions with high risk human papillomavirus infection. Int J. Clin. Exp. Med. 8, 21080–21088 (2015).

pubmed: 26885039 pmcid: 4723884

Moscicki, A. B., Shi, B., Huang, H., Barnard, E. & Li, H. Cervical-vaginal microbiome and associated cytokine profiles in a prospective study of HPV 16 acquisition, persistence, and clearance. Front Cell Infect. Microbiol. 10, 569022 (2020).

pubmed: 33102255 pmcid: 7546785 doi: 10.3389/fcimb.2020.569022

Cheng, L. et al. Vaginal microbiota and human papillomavirus infection among young Swedish women. NPJ Biofilms Microbiomes 6, 39 (2020).

pubmed: 33046723 pmcid: 7552401 doi: 10.1038/s41522-020-00146-8

Lee, J. E. et al. Association of the vaginal microbiota with human papillomavirus infection in a Korean twin cohort. PLoS ONE 8, e63514 (2013).

pubmed: 23717441 pmcid: 3661536 doi: 10.1371/journal.pone.0063514

Mitra, A. et al. Cervical intraepithelial neoplasia disease progression is associated with increased vaginal microbiome diversity. Sci. Rep. 5, 16865 (2015).

pubmed: 26574055 pmcid: 4648063 doi: 10.1038/srep16865

Perkins, R. B. et al. 2019 ASCCP risk-based management consensus guidelines for abnormal cervical cancer screening tests and cancer precursors. J. Low. Genit. Trac. Dis. 24, 102–131 (2020).

doi: 10.1097/LGT.0000000000000525

DasGupta, T. et al. Human papillomavirus oncogenic E6 protein regulates human beta-defensin 3 (hBD3) expression via the tumor suppressor protein p53. Oncotarget 7, 27430–27444 (2016).

pubmed: 27034006 pmcid: 5053661 doi: 10.18632/oncotarget.8443

Nguyen, H., Teskey, L., Lin, R. & Hiscott, J. Identification of the secretory leukocyte protease inhibitor (SLPI) as a target of IRF-1 regulation. Oncogene 18, 5455–5463 (1999).

pubmed: 10498899 doi: 10.1038/sj.onc.1202924

Um, S. J. et al. Abrogation of IRF-1 response by high-risk HPV E7 protein in vivo. Cancer Lett. 179, 205–212 (2002).

pubmed: 11888675 doi: 10.1016/S0304-3835(01)00871-0

Hasan, U. A. et al. The human papillomavirus type 16 E7 oncoprotein induces a transcriptional repressor complex on the Toll-like receptor 9 promoter. J. Exp. Med. 210, 1369–1387 (2013).

pubmed: 23752229 pmcid: 3698525 doi: 10.1084/jem.20122394

Suarez-Carmona, M., Hubert, P., Delvenne, P. & Herfs, M. Defensins: “Simple” antimicrobial peptides or broad-spectrum molecules? Cytokine growth factor Rev. 26, 361–370 (2015).

pubmed: 25578522 doi: 10.1016/j.cytogfr.2014.12.005

Hancock, R. E., Haney, E. F. & Gill, E. E. The immunology of host defence peptides: beyond antimicrobial activity. Nat. Rev. Immunol. 16, 321–334 (2016).

pubmed: 27087664 doi: 10.1038/nri.2016.29

Hubert, P. et al. Defensins induce the recruitment of dendritic cells in cervical human papillomavirus-associated (pre)neoplastic lesions formed in vitro and transplanted in vivo. FASEB J.: Off. Publ. Federation Am. Societies Exp. Biol. 21, 2765–2775 (2007).

doi: 10.1096/fj.06-7646com

Hubert, P. et al. Altered alpha-defensin 5 expression in cervical squamocolumnar junction: implication in the formation of a viral/tumour-permissive microenvironment. J. Pathol. 234, 464–477 (2014).

pubmed: 25196670 doi: 10.1002/path.4435

Wolf, R. et al. Chemotactic activity of S100A7 (Psoriasin) is mediated by the receptor for advanced glycation end products and potentiates inflammation with highly homologous but functionally distinct S100A15. J. Immunol. 181, 1499–1506 (2008).

pubmed: 18606705 doi: 10.4049/jimmunol.181.2.1499

Presicce, P., Giannelli, S., Taddeo, A., Villa, M. L. & Della Bella, S. Human defensins activate monocyte-derived dendritic cells, promote the production of proinflammatory cytokines, and up-regulate the surface expression of CD91. J. Leukoc. Biol. 86, 941–948 (2009).

pubmed: 19477909 doi: 10.1189/jlb.0708412

Wiens, M. E. & Smith, J. G. Alpha-defensin HD5 inhibits furin cleavage of human papillomavirus 16 L2 to block infection. J. Virol. 89, 2866–2874 (2015).

pubmed: 25540379 doi: 10.1128/JVI.02901-14

Wiens, M. E. & Smith, J. G. alpha-Defensin HD5 inhibits human papillomavirus 16 infection via capsid stabilization and redirection to the lysosome. mBio 8, e02304-16 (2017).

Raveschot, C. et al. Production of bioactive peptides by lactobacillus species: from gene to application. Front. Microbiol. 9, 2354 (2018).

pubmed: 30386307 pmcid: 6199461 doi: 10.3389/fmicb.2018.02354

Sadat-Mekmene, L., Genay, M., Atlan, D., Lortal, S. & Gagnaire, V. Original features of cell-envelope proteinases of Lactobacillus helveticus: a review. Int. J. Food Microbiol. 146, 1–13 (2011).

pubmed: 21354644 doi: 10.1016/j.ijfoodmicro.2011.01.039

Rollison, D. E., Viarisio, D., Amorrortu, R. P., Gheit, T. & Tommasino, M. An emerging issue in oncogenic virology: the role of beta human papillomavirus types in the development of cutaneous squamous cell carcinoma. J. Virol. 93, e01003–18 (2019).

Tummers, B. et al. The interferon-related developmental regulator 1 is used by human papillomavirus to suppress NFkappaB activation. Nat. Commun. 6, 6537 (2015).

pubmed: 26055519 doi: 10.1038/ncomms7537

Schrofelbauer, B., Polley, S., Behar, M., Ghosh, G. & Hoffmann, A. NEMO ensures signaling specificity of the pleiotropic IKKbeta by directing its kinase activity toward IkappaBalpha. Mol. Cell 47, 111–121 (2012).

pubmed: 22633953 pmcid: 3398199 doi: 10.1016/j.molcel.2012.04.020

Griffiths, D. A. et al. Merkel cell polyomavirus small T antigen targets the NEMO adaptor protein to disrupt inflammatory signaling. J. Virol. 87, 13853–13867 (2013).

pubmed: 24109239 pmcid: 3838273 doi: 10.1128/JVI.02159-13

Cicchini, L. et al. High-risk human papillomavirus E7 alters host DNA methylome and represses HLA-E expression in human keratinocytes. Sci. Rep. 7, 3633 (2017).

pubmed: 28623356 pmcid: 5473897 doi: 10.1038/s41598-017-03295-7

Lee, J. O. et al. Hepatitis B virus X protein represses E-cadherin expression via activation of DNA methyltransferase 1. Oncogene 24, 6617–6625 (2005).

pubmed: 16007161 doi: 10.1038/sj.onc.1208827

Kanarek, N. & Ben-Neriah, Y. Regulation of NF-kappaB by ubiquitination and degradation of the IkappaBs. Immunological Rev. 246, 77–94 (2012).

doi: 10.1111/j.1600-065X.2012.01098.x

Liu, C. et al. beta-Trcp couples beta-catenin phosphorylation-degradation and regulates Xenopus axis formation. Proc. Natl Acad. Sci. USA 96, 6273–6278 (1999).

pubmed: 10339577 pmcid: 26871 doi: 10.1073/pnas.96.11.6273

Busino, L. et al. Degradation of Cdc25A by beta-TrCP during S phase and in response to DNA damage. Nature 426, 87–91 (2003).

pubmed: 14603323 doi: 10.1038/nature02082

Ison, C. A. & Hay, P. E. Validation of a simplified grading of Gram stained vaginal smears for use in genitourinary medicine clinics. Sexually transmitted Infect. 78, 413–415 (2002).

doi: 10.1136/sti.78.6.413

Chawla, R., Bhalla, P., Chadha, S., Grover, S. & Garg, S. Comparison of Hay’s criteria with Nugent’s scoring system for diagnosis of bacterial vaginosis. BioMed. Res. Int. 2013, 365194 (2013).

pubmed: 23841066 pmcid: 3697286 doi: 10.1155/2013/365194

Bhujel, R., Mishra, S. K., Yadav, S. K., Bista, K. D. & Parajuli, K. Comparative study of Amsel’s criteria and Nugent scoring for diagnosis of bacterial vaginosis in a tertiary care hospital, Nepal. BMC Infect. Dis. 21, 825 (2021).

pubmed: 34404367 pmcid: 8369704 doi: 10.1186/s12879-021-06562-1

Sha, B. E. et al. Utility of Amsel criteria, Nugent score, and quantitative PCR for Gardnerella vaginalis, Mycoplasma hominis, and Lactobacillus spp. for diagnosis of bacterial vaginosis in human immunodeficiency virus-infected women. J. Clin. Microbiol. 43, 4607–4612 (2005).

pubmed: 16145114 pmcid: 1234056 doi: 10.1128/JCM.43.9.4607-4612.2005

Arbeit, J. M., Howley, P. M. & Hanahan, D. Chronic estrogen-induced cervical and vaginal squamous carcinogenesis in human papillomavirus type 16 transgenic mice. Proc. Natl Acad. Sci. USA 93, 2930–2935 (1996).

pubmed: 8610145 pmcid: 39737 doi: 10.1073/pnas.93.7.2930

Elson, D. A. et al. Sensitivity of the cervical transformation zone to estrogen-induced squamous carcinogenesis. Cancer Res. 60, 1267–1275 (2000).

pubmed: 10728686

Gerard, C. et al. Accurate Control of 17beta-estradiol long-term release increases reliability and reproducibility of preclinical animal studies. J. mammary gland Biol. neoplasia 22, 1–11 (2017).

pubmed: 27889857 doi: 10.1007/s10911-016-9368-1

Goldman, J. M., Murr, A. S. & Cooper, R. L. The rodent estrous cycle: characterization of vaginal cytology and its utility in toxicological studies. Birth defects Res. Part B, Dev. Reprod. Toxicol. 80, 84–97 (2007).

doi: 10.1002/bdrb.20106

Wimmer-Scherr, C., et al. Comparison of fecal microbiota of horses suffering from atypical myopathy and healthy co-grazers. Animals (Basel) 11, 506 (2021).

Cerri, S. et al. Effect of oral administration of omeprazole on the microbiota of the gastric glandular mucosa and feces of healthy horses. J. Vet. Intern Med. 34, 2727–2737 (2020).

pubmed: 33063923 pmcid: 7694827 doi: 10.1111/jvim.15937

Fettweis, G. et al. RIP3 antagonizes a TSC2-mediated pro-survival pathway in glioblastoma cell death. Biochimica et. Biophysica Acta Mol. Cell Res. 1864, 113–124 (2017).

doi: 10.1016/j.bbamcr.2016.10.014

Blomme, A. et al. Myoferlin regulates cellular lipid metabolism and promotes metastases in triple-negative breast cancer. Oncogene 36, 2116–2130 (2017).

pubmed: 27775075 doi: 10.1038/onc.2016.369

Lambert, P. F. et al. Using an immortalized cell line to study the HPV life cycle in organotypic “raft” cultures. Methods Mol. Med. 119, 141–155 (2005).

pubmed: 16353335

Meuris, F. et al. The CXCL12/CXCR4 signaling pathway: a new susceptibility factor in human papillomavirus pathogenesis. PLoS Pathog. 12, e1006039 (2016).

pubmed: 27918748 pmcid: 5138052 doi: 10.1371/journal.ppat.1006039

Nys, G., Cobraiville, G. & Fillet, M. Multidimensional performance assessment of micro pillar array column chromatography combined to ion mobility-mass spectrometry for proteome research. Analytica Chim. Acta 1086, 1–13 (2019).

doi: 10.1016/j.aca.2019.08.068

Herfs, M. et al. A dualistic model of primary anal canal adenocarcinoma with distinct cellular origins, etiologies, inflammatory microenvironments and mutational signatures: implications for personalised medicine. Br. J. Cancer 118, 1302–1312 (2018).

pubmed: 29700411 pmcid: 5959925 doi: 10.1038/s41416-018-0049-2

Herfs, M. et al. A novel blueprint for ‘top down’ differentiation defines the cervical squamocolumnar junction during development, reproductive life, and neoplasia. J. Pathol. 229, 460–468 (2013).

pubmed: 23007879 doi: 10.1002/path.4110

Herfs, M. et al. Transforming growth factor-beta1-mediated Slug and Snail transcription factor up-regulation reduces the density of Langerhans cells in epithelial metaplasia by affecting E-cadherin expression. Am. J. Pathol. 172, 1391–1402 (2008).

pubmed: 18385519 pmcid: 2329847 doi: 10.2353/ajpath.2008.071004

Wang, X., Spandidos, A., Wang, H. & Seed, B. PrimerBank: a PCR primer database for quantitative gene expression analysis, 2012 update. Nucleic Acids Res. 40, D1144–D1149 (2012).

pubmed: 22086960 doi: 10.1093/nar/gkr1013

Poirson, J. et al. Mapping the interactome of HPV E6 and E7 oncoproteins with the ubiquitin-proteasome system. FEBS J. 284, 3171–3201 (2017).

pubmed: 28786561 doi: 10.1111/febs.14193

Cassonnet, P. et al. Benchmarking a luciferase complementation assay for detecting protein complexes. Nat. Methods 8, 990–992 (2011).

pubmed: 22127214 doi: 10.1038/nmeth.1773

Nokin, M. J., et al. Methylglyoxal, a glycolysis side-product, induces Hsp90 glycation and YAP-mediated tumor growth and metastasis. eLife 5, e19375 (2016).