A Human Papillomavirus Early Promoter Minimal Model: Viral Population and Stochasticity

ObjectiveAim of this study was to assess the changes in genetic and epigenetic profiles of human papillomavirus type 16 (HPV16), if any, in primary cervical cancer (CaCx) and corresponding plasma before and after therapy for possible prognostic evaluation.MethodsThe genetic (integration status) and epigenetic (methylation of enhancer, early promoter, and late promoter sequences) profiles of HPV16 were analyzed in pretherapy CaCx (n = 46), corresponding plasma, posttherapy cervical swabs (n = 39), and corresponding plasma from a single patient cohort. Quantitative viral load was also measured in these HPV16-positive primary CaCx and posttherapy cervical swabs.ResultsPresence of HPV16 in the patients’ plasma before/after therapy was significantly (P= 0.03) associated with higher viral load in the primary tumor site. Human papillomavirus type 16 integration and hypomethylation of the early (14 of 29,Z= 4.47,P< 0.01) and late promoters (20 of 29,Z= 3.74,P< 0.01) were more prevalent in the plasma than the corresponding pretherapy CaCx samples. However, the dissimilarity in integration status (5 of 24) was less evident between posttherapy cervical swabs and corresponding plasma, although hypomethylation of the early promoter and hypermethylation of the late promoter (8 of 24,Z= 2.6,P< 0.01) was seen in posttherapy plasma samples. Whereas in the posttherapy swabs, integrated (22 of 29) or mixed (7 of 29) form of HPV16 prevailed with hypomethylation of the enhancer (6 of 29,Z= 2.0,P< 0.05) and late promoter (18 of 29,Z= 4.4,P< 0.01) compared with the corresponding primary tumors. The patients having high HPV16 copy number in pretherapy and posttherapy cervical lesions and hypomethylation of early promoter/late promoter in the corresponding plasma showed increased disease recurrence with distant metastases.ConclusionsThe genetic-epigenetic profile of HPV16 in pretherapy/posttherapy CaCx samples showed significant association with disease prognosis.

Download Full-text

Human papillomavirus 16 L1 gene methylation as a potential biomarker for predicting anal intraepithelial neoplasia in men who have sex with men (MSM)

PLoS ONE ◽

10.1371/journal.pone.0256852 ◽

2021 ◽

Vol 16 (9) ◽

pp. e0256852

Author(s):

Arkom Chaiwongkot ◽

Nittaya Phanuphak ◽

Tippawan Pankam ◽

Parvapan Bhattarakosol

Keyword(s):

Human Papillomavirus ◽

Methylation Status ◽

Intraepithelial Neoplasia ◽

Anal Intraepithelial Neoplasia ◽

Gene Methylation ◽

Human Papillomavirus 16 ◽

Early Promoter ◽

Potential Biomarker ◽

Sex With Men ◽

Methylation Patterns

The human papillomavirus (HPV) 16 early promoter and L1 gene methylation were quantitatively measured using pyrosequencing assay in anal cells collected from men who have sex with men (MSM) to determine potential biomarkers for HPV-related anal cancer. The methylation patterns of HPV16 genes, including the early promoter (CpG 31, 37, 43, 52, and 58) and L1 genes (CpG 5600, 5606, 5609, 5615, 7136, and 7145), were analyzed in 178 anal samples. The samples were diagnosed as normal, anal intraepithelial neoplasia (AIN) 1, AIN2, and AIN3. Low methylation levels of the early promoter (< 10%) and L1 genes (< 20%) were found in all detected normal anal cells. In comparison, medium to high methylation (≥ 20–60%) in the early promoter was found in 1.5% (1/67) and 5% (2/40) of AIN1 and AIN2-3 samples, respectively. Interestingly, slightly increased L1 gene methylation levels (≥ 20–60%), especially at the HPV16 5’L1 regions CpGs 5600 and 5609, were demonstrated in AIN2-3 specimen. Moreover, a negative correlation between high HPV16 L1 gene methylation at CpGs 5600, 5609, 5615, and 7145 and a percentual CD4 count was found in AIN3 HIV positive cases. When comparing the methylation status of AIN2-3 to that of normal/AIN1 lesions, the results indicated the potential of using HPV16 L1 gene methylation as a biomarker for HPV-related cancer screening.

Download Full-text

In Vivo Tissue-Specific Regulation of the Human Papillomavirus Type 18 Early Promoter by Estrogen, Progesterone, and Their Antagonists

Virology ◽

10.1006/viro.2001.1287 ◽

2002 ◽

Vol 294 (1) ◽

pp. 135-140 ◽

Cited By ~ 6

Author(s):

Néstor Morales-Peza ◽

Prasert Auewarakul ◽

Victoria Juárez ◽

Alejandro Garcı́a-Carrancá ◽

Angel Cid-Arregui

Keyword(s):

Human Papillomavirus ◽

Tissue Specific ◽

Early Promoter ◽

Specific Regulation

Download Full-text

The Transcriptional Cofactor VGLL1 Drives Transcription of Human Papillomavirus Early Genes via TEAD1

Journal of Virology ◽

10.1128/jvi.01945-19 ◽

2020 ◽

Vol 94 (10) ◽

Author(s):

Seiichiro Mori ◽

Takamasa Takeuchi ◽

Yoshiyuki Ishii ◽

Iwao Kukimoto

Keyword(s):

Gene Expression ◽

Cervical Cancer ◽

Transcription Factors ◽

Human Papillomavirus ◽

Early Gene ◽

Luciferase Reporter ◽

Early Gene Expression ◽

Early Genes ◽

Early Promoter

ABSTRACT The TEAD family of transcription factors requires associating cofactors to induce gene expression. TEAD1 is known to activate the early promoter of human papillomavirus (HPV), but the precise mechanisms of TEAD1-mediated transactivation of the HPV promoter, including its relevant cofactors, remain unexplored. Here, we reveal that VGLL1, a TEAD-interacting cofactor, contributes to HPV early gene expression. Knockdown of VGLL1 and/or TEAD1 led to a decrease in viral early gene expression in human cervical keratinocytes and cervical cancer cell lines. We identified 11 TEAD1 target sites in the HPV16 long control region (LCR) by in vitro DNA pulldown assays; 8 of these sites contributed to the transcriptional activation of the early promoter in luciferase reporter assays. VGLL1 bound to the HPV16 LCR via its interaction with TEAD1 both in vitro and in vivo. Furthermore, introducing HPV16 and HPV18 whole genomes into primary human keratinocytes led to increased levels of VGLL1, due in part to the upregulation of TEADs. These results suggest that multiple VGLL1/TEAD1 complexes are recruited to the LCR to support the efficient transcription of HPV early genes. IMPORTANCE Although a number of transcription factors have been reported to be involved in HPV gene expression, little is known about the cofactors that support HPV transcription. In this study, we demonstrate that the transcriptional cofactor VGLL1 plays a prominent role in HPV early gene expression, dependent on its association with the transcription factor TEAD1. Whereas TEAD1 is ubiquitously expressed in a variety of tissues, VGLL1 displays tissue-specific expression and is implicated in the development and differentiation of epithelial lineage tissues, where HPV gene expression occurs. Our results suggest that VGLL1 may contribute to the epithelial specificity of HPV gene expression, providing new insights into the mechanisms that regulate HPV infection. Further, VGLL1 is also critical for the growth of cervical cancer cells and may represent a novel therapeutic target for HPV-associated cancers.

Download Full-text

Potential biomarker of human papillomavirus 16 L1 methylation for prediction of anal intraepithelial neoplasia in men who have sex with men (MSM)

10.1101/2021.01.07.425707 ◽

2021 ◽

Author(s):

Arkom Chaiwongkot ◽

Parvapan Bhattarakosol ◽

Nittaya Phanuphak ◽

Tippawan Pankam

Keyword(s):

Human Papillomavirus ◽

Methylation Status ◽

Intraepithelial Neoplasia ◽

Anal Intraepithelial Neoplasia ◽

Gene Methylation ◽

Human Papillomavirus 16 ◽

Early Promoter ◽

Potential Biomarker ◽

Sex With Men ◽

Methylation Patterns

Quantitative measurement of human papillomavirus (HPV) 16 early promoter and L1 genes methylation were analyzed in anal cells collected from men who have sex with men (MSM) to determine the potential biomarker for screening of HPV related anal cancer. The methylation patterns of the HPV16 genes including early promoter (CpG 31, 37, 43, 52 and 58) and L1 gene (CpG 5600, 5606, 5609, 5615, 7136 and 7145) were analyzed in 178 anal samples with histology diagnosed as normal, anal intraepithelial neoplasia (AIN) 1, AIN2 and AIN3 by pyrosequencing assay. Low methylation levels of early promoter (<10%) and L1 genes (<20%) were found in all detected normal anal cells, while medium to high methylation (>20-60%) in early promoter was found 1.5% (1/67) and 5% (2/40) in AIN1 and AIN2-3 samples, respectively. Interestingly, slightly increased L1 gene methylation level (>20-60%) especially at HPV16 5'L1 regions CpGs 5600 and 5609 from normal to AIN3 were demonstrated. Moreover, negative correlation between high HPV16 L1 gene methylation at CpGs 5600, 5609, 5615 and 7145 and low percentage of CD4+ was found in AIN3 HIV positive cases. When compared methylation status of AIN2-3 to those of the normal/AIN1 lesion, the results indicated potential of using HPV16 L1 gene methylation as a biomarker for HPV related cancer screening.

Download Full-text