Diagnosing Cervical Cancer and High-Grade Precursors by HPV16 Transcription Patterns

Schmitt, Markus; Dalstein, Véronique; Waterboer, Tim; Clavel, Christine; Gissmann, Lutz; Pawlita, Michael

doi:10.1158/0008-5472.can-09-2514

Cited by 69 publications

(76 citation statements)

References 31 publications

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“…In practice, our data open possibilities to test women in primary screening with a validated quantitative HPV assay that preferably detects all Hr-HPV types. For those that test positive for high viral loads, reflex cytology or HPV RNA pattern detection (32) could be conducted to identify women with abnormal cytology who need direct referral for colposcopydirected biopsy. In this way, up to 81% of Hr-HPV positive but healthy women could be saved from extensive follow-up, which would result in a significant increase in specificity and PPV of Hr-HPV testing for HSIL and CIN2þ in cervical screening programs.…”

Section: Discussionmentioning

confidence: 99%

Viral Load of High-Risk Human Papillomaviruses as Reliable Clinical Predictor for the Presence of Cervical Lesions

Schmitt

Depuydt

Benoy

et al. 2013

Cancer Epidemiology, Biomarkers &Amp; Prevention

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Background: Infections with high-risk human papillomaviruses (Hr-HPV) can cause malignant transformation of the human cervical epithelium. HPV DNA tests generally are very sensitive to detect cervical neoplastic lesions but also identify transient HPV infections. As a consequence, the specificity and positive predictive value are low.Methods: We analyzed viral load of Hr-and possibly Hr-HPV types more than seven orders of magnitude (on a log10 scale) in 999 consecutive BD-SurePath liquid-based cervical cytology samples from routine cervical screening enriched with atypical squamous cells of undetermined significance (n ¼ 100), low-grade squamous intraepithelial lesions (LSIL; n ¼ 100), and high-grade squamous intraepithelial lesions (HSIL; n ¼ 97) using type-specific multiplex quantitative real-time PCR and the BSGP5þ/6þ-PCR/MPG assay. In the 36-month follow-up, 79 histologically verified CIN2þ and 797 double-negative cytology cases were identified.Results: Viral loads in LSIL and HSIL were significantly increased compared with no intraepithelial lesion or malignancy in both the quantitative PCR (qPCR) and BSGP5þ/6þ-PCR/MPG assay (P < 0.0001). The mean viral loads in LSIL and HSIL were not significantly different. Using a newly determined high viral load cut off for 14 Hr-HPV types, the sensitivity for prevalent CIN3þ remained at 100% for both assays compared with the minimal detection threshold. The specificity (corresponding to double-negative cytology at subsequent screening episodes) increased substantially (qPCR, from 91.1% to 95.7%; BSGP5þ/6þ-PCR/MPG, from 79.8% to 96.2%).Conclusions: Compared with DNA positivity alone, high Hr-HPV viral loads could reduce the amount of false positive results detected by the BSGP5þ/6þ-PCR/MPG and qPCR by 81.4% and 52.1%, respectively.Impact: Quantitative type-specific HPV DNA assays show high flexibility in defining thresholds that allow optimizing clinical accuracy for cervical cancer precursors. Cancer Epidemiol Biomarkers Prev; 22(3); 406-14. Ó2013 AACR.

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Section: Discussionmentioning

confidence: 99%

Viral Load of High-Risk Human Papillomaviruses as Reliable Clinical Predictor for the Presence of Cervical Lesions

Schmitt

Depuydt

Benoy

et al. 2013

Cancer Epidemiology, Biomarkers &Amp; Prevention

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“…1A) (6). Some of these isoforms have been detected previously in cell lines and patient tissues (7,12,(40)(41)(42)(43)(44). However, it is unclear whether all isoforms can be expressed simultaneously and/or are differentially expressed in nontumor versus tumor cells.…”

Section: Hpv16mentioning

confidence: 98%

Human Papillomavirus 16 Oncoprotein Expression Is Controlled by the Cellular Splicing Factor SRSF2 (SC35)

McFarlane

MacDonald

Stevenson

et al. 2015

J Virol

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High-risk human papillomaviruses (HR-HPV) cause anogenital cancers, including cervical cancer, and head and neck cancers. Human papillomavirus 16 (HPV16) is the most prevalent HR-HPV. HPV oncogenesis is driven by two viral oncoproteins, E6 and E7, which are expressed through alternative splicing of a polycistronic RNA to yield four major splice isoforms (E6 full length, E6*I, E6*II, E6*X). The production of multiple mRNA isoforms from a single gene is controlled by serine/arginine-rich splicing factors (SRSFs), and HPV16 infection induces overexpression of a subset of these, SRSFs 1, 2, and 3. In this study, we examined whether these proteins could control HPV16 oncoprotein expression. Small interfering RNA (siRNA) depletion experiments revealed that SRSF1 did not affect oncoprotein RNA levels. While SRSF3 knockdown caused some reduction in E6E7 expression, depletion of SRSF2 resulted in a significant loss of E6E7 RNAs, resulting in reduced levels of the E6-regulated p53 proteins and E7 oncoprotein itself. SRSF2 contributed to the tumor phenotype of HPV16-positive cervical cancer cells, as its depletion resulted in decreased cell proliferation, reduced colony formation, and increased apoptosis. SRSF2 did not affect transcription from the P 97 promoter that controls viral oncoprotein expression. Rather, RNA decay experiments showed that SRSF2 is required to maintain stability of E6E7 mRNAs. These data show that SRSF2 is a key regulator of HPV16 oncoprotein expression and cervical tumor maintenance. H uman papillomaviruses (HPV) infect mucosal and cutaneous epithelia. At least 13 so-called "high-risk" HPV (HR-HPV) infect the anogenital epithelium and can cause persistent lesions that may progress to cancer (1). For example, around 500,000 women worldwide experience anogenital HPV infection, and nearly 300,000 die per annum from cervical cancer. Increasingly, HPV infection is also being linked to oropharyngeal cancer, whereby incidence of this disease is increasing rapidly (2). HPV16 is the most prevalent HR-HPV. HPV-associated tumorigenesis is driven by increased expression of the HPV E6 and E7 oncoproteins (3). E6 promotes ubiquitin-mediated degradation of p53 to inhibit apoptosis, modulates transcription of cell cycle-related genes, induces telomerase activity, controls cell shape and polarity, and activates cap-dependent translation (4). E7 binds and degrades Rb to promote S phase entry and cell division, controls transcription of cell cycle-related genes, and acts as a mitotic mutator (4). IMPORTANCE Expression of the HPV16 oncoproteins E7 and E6 drives HPV-associated tumor formation. Although increased transcription may yield increased levels of E6E7 mRNAs, it is known that theHPV E6 and E7 oncoproteins are expressed from a polycistronic transcript that for HPV16 can potentially produce four different alternatively spliced mRNAs (E6 full length [E6fl], E6*I, E6*II, and E6*X [also called E6*III]) (5, 6). The putative E6* proteins all share the first 44 amino acids of full-length E6 with C-terminal trun...

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“…DNase I digestion (Qiagen) was included to ensure an exclusive amplification of RNA. Nucleic acid sequence-based amplification and hybridization to splice-specific probes on Luminex beads were carried out as described (30 …”

Section: Patients Tissue Samplesmentioning

confidence: 99%

Viral RNA Patterns and High Viral Load Reliably Define Oropharynx Carcinomas with Active HPV16 Involvement

et al. 2012

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Oropharyngeal squamous cell carcinomas (OPSCC) that are associated with human papilloma virus (HPV) infection carry a more favorable prognosis than those that are HPV-negative. However, it remains unclear which biomarker(s) can reliably determine which OPSCC specimens are truly driven by HPV infection. In this study, we analyzed 199 fresh-frozen OPSCC specimens for HPV DNA, viral load, RNA expression patterns typical for cervical carcinomas (CxCaRNA þ ), and the HPV-targeted tumor suppressor protein p16 INK4a as markers for HPV infection.In this set of specimens, there was a 49% prevalence of DNA for the cancer-associated HPV type 16 (HPV þ ).

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Diagnosing Cervical Cancer and High-Grade Precursors by HPV16 Transcription Patterns

Cited by 69 publications

References 31 publications

Viral Load of High-Risk Human Papillomaviruses as Reliable Clinical Predictor for the Presence of Cervical Lesions

Viral Load of High-Risk Human Papillomaviruses as Reliable Clinical Predictor for the Presence of Cervical Lesions

Human Papillomavirus 16 Oncoprotein Expression Is Controlled by the Cellular Splicing Factor SRSF2 (SC35)

Viral RNA Patterns and High Viral Load Reliably Define Oropharynx Carcinomas with Active HPV16 Involvement

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