HPV Positivity Research Articles

Personalized oncogenomic analysis of metastatic squamous cell carcinoma of the anus: Utilizing whole-genome sequencing to guide clinical decision-making.

11 Background: Metastatic squamous cell carcinoma of the anus (SCCA) is associated with significant morbidity and mortality. Due to its relative rarity, there is limited evidence to support systemic therapy regimens informed by genomic data. This study aims to utilize whole-genome and transcriptome sequencing to enhance the understanding of the genetic alterations driving metastatic SCCA and to identify potentially actionable therapeutic targets. Methods: This report examines nine cases of metastatic SCCA in patients enrolled in the Personalized Oncogenomics (POG) Program at the BC Cancer Agency. Comprehensive genomic profiling, including whole-genome and transcriptome analysis (WGTA) was conducted on fresh tumor biopsy or formalin fixed paraffin embedded (FFPE) tissue. Tumor genomic alterations were analyzed in detail including: SNVs, indels, copy number alterations, structural variants, mutation signatures, viral presence, tumor mutation burden (TMB), expression outliers, and immune cell scores. The somatic alterations were integrated with existing knowledge of drug-target interactions to identify actionable therapeutic targets. Results: Of the nine patients, eight were found to be HPV-positive, with one exhibiting high tumor mutational burden (TMB>10mut/Mb). Mutation signatures included seven cases with AID/APOBEC signatures (commonly seen with HPV), and one case with an ID2 DNA replication slippage signature. The PI3K/AKT/mTOR pathway was the most commonly affected signaling pathway (n=4), followed by FGFR amplification and overexpression (n=3), and RAS/RAF alterations ( BRAF mutation and KRAS overexpression, n=2). EGFR amplification was identified in two cases, one of which also exhibited EGFR overexpression. Additionally, FBXW7 mutations were present in two cases. Immunotherapy was recommended for all nine patients: eight based on HPV positivity, either alone or in combination with other alterations, such as high TMB, SWI/SNF complex alterations, and presence of immune infiltrating cells, and the remaining case based on a SWI/SNF complex mutation. Only two patients ultimately received immunotherapy, both of whom derived clinical benefit. One patient, treated with atezolizumab as part of the phase II CAPTIV-8 trial (NCT04273061), demonstrated progression-free survival (PFS) of 10 months (ongoing), while the other, treated with nivolumab, achieved a PFS of 6 months. Conclusions: Genomic analysis using WGTA supported the recommendation of immunotherapy for all patients in this cohort. Although only two patients received immunotherapy, both experienced clinical benefit, underscoring the potential of personalized genomic-guided treatment in metastatic SCCA.

Management of women with abnormal cervical cytology: Update of INCa recommendations after the implementation of HPV screening

To update the recommendations issued by the National Cancer Institute (INCa) on the management of women with abnormal cervical cytology. INCa recommendations on the management of women with abnormal cervical cytology were published in 2016. In 2019, the High Authority of Health (HAS) recommends HPV test for cervical cancer screening in women over 30. It was therefore necessary to update the initial management and follow-up of women with a reflex cytology abnormality after a positive HPV test. We used the modified Delphi method. Each decision tree was modified by the 3 members of the organizing committee and then presented to 35 experts over 2 rounds, in order to obtain their agreement in terms of validity and clarity. This formalized consensus of experts allowed the creation of 17 decision trees instead of 15 previously: 13 on the diagnostic management of abnormal cytology, stratified according to age, and 4 on therapeutic indications in abnormal histology. The median agreement scores were 89% (81-97). These 17 decision trees respond to the different situations that health professionals may encounter in order to enable them to act, to follow-up or delegate according to their specialties.

Risk of residual/recurrent cervical diseases in HPV-positive women post-conization depends on HPV integration status

BackgroundIt is crucial to identify post-operative patients with HPV infection who are at high risk for residual/recurrent disease. This study aimed to evaluate the association between HPV integration and clinical outcomes in HPV-positive women after cervical conization, as well as to identify HPV integration breakpoints.MethodsThis retrospective study analyzed data of 791 women who underwent cervical conization for cervical intraepithelial neoplasia grades 2–3 (CIN2-3) between September 2019 and September 2023, sourced from the Fujian and Hubei cervical lesion screening cohorts. Among these, 73 women with HPV infection post-conization underwent HPV integration test within 3 months after a positive HPV test. HPV integration test was performed using the high-throughput viral integration detection (HIVID), a sensitive method for genome-wide survey of HPV integration breakpoints.ResultsAmong the 73 participants with HPV infection post-conization, 10 cases (13.7%) were positive for HPV integration. The logistic regression analysis showed a higher residual/recurrent lesions risk in patients with HPV integration (OR = 3.917, p = 0.048). According to the Kaplan-Meier analysis, age ≥ 45 years (p = 0.016) and HPV integration (p = 0.035) were associated with a higher risk of residual/recurrent CIN at the 1-year follow-up. HPV 52 accounted for the majority of HPV integration genotype (3/10, 30.0%). Surprisingly, HPV 16 had the highest number of HPV average integration sequencing reads (n = 129), followed by HPV 31, 58, 52, 59, 35, and 39. The study also identified 13 HPV breakpoints, including TP63, TLR4, USP10, etc.ConclusionsHPV integration was identified as an independent risk factor for residual/recurrent CIN in HPV-positive women post-conization. Women with positive HPV integration should pay attention to careful post-treatment follow-up.

Impact of Smoking on Cervical Histopathological Changes in High-Risk HPV-Positive Women: A Matched Case–Control Study

Background and Objectives: The aims of this study were to assess the impact of smoking on cervical histopathology in women with high-risk HPV types 16 and 18 (the most common types) utilizing comprehensive clinical data and to conduct a risk analysis based on smoking pack-years. Materials and Methods: Between 2022 and 2024, 1048 high-risk HPV-positive women aged 25 to 65 years were categorized into two groups: smokers and non-smokers. Data acquired from a histopathological examination of samples collected during a colposcopic evaluation of these women were compared individually regarding clinical and demographic factors, specifically age, gravida, parity, and alcohol consumption. Subsequently, the impact of prolonged and excessive smoking on histopathological cellular changes was assessed in women with the same characteristics. A case–control study was performed on 312 smokers and 312 non-smokers following mutual matching. Results: The women were matched one-to-one regarding gravida, parity, and alcohol consumption. Subsequently, they were paired within a ±2-year age range. The mean age of the smoker group was 47.1 ± 8.8, while that of the non-smoker group was 47.2 ± 8.5 (p: 0.904). In all cases of high-risk HPV positivity, the rate of normal cervical cytological results was 14% in women who smoked and 29% in women who did not smoke. The LGSIL, HGSIL, ASC-H, and AGC-NOS rates were elevated in the smoker group, and a statistically significant difference was observed between the two groups in terms of abnormal cervical cytological results (p < 0.001). After a colposcopic biopsy, the smoker group exhibited higher rates of HGSILs, LGSILs, AGC-NOS, and CIS pathological lesions (28% vs. 23%), whereas the non-smoker group exhibited higher rates of chronic cervicitis (23% vs. 16%). However, no statistically significant difference was found between the two groups (p: 0.092). In a comparison of endocervical curettage (ECC) samples, it was observed that the HGSIL, CIS, and AGC-FN rates in the smoker group were almost the same as those in the non-smoker group. However, the LGSIL histopathology results (32% vs. 18%) were higher, and the rate of negativity with no pathology was higher in the non-smoker group (72% vs. 59%). A statistically significant difference in ECC histopathology was noted between the two groups (p < 0.001). An ROC analysis conducted between smoking pack-years and the colposcopic and endocervical curettage biopsy results revealed that the cutoff value for the colposcopic abnormal histopathological results increased, with 40% sensitivity and 76% specificity above 20 pack-years (AUC: 0.592 and p: 0.025). Additionally, the abnormal histopathology rates for endocervical curettage exhibited 81% sensitivity and 32% specificity above 13 pack-years (AUC: 0.586 and p: 0.008). The rate of abnormalities in the colposcopic biopsy results was 2.19 times higher for individuals with over 20 pack-years, and the rate of abnormalities in the ECC results was 2.08 times higher for those with over 13 pack-years; additionally, statistically significant results were obtained (p-values of 0.027 and 0.008, respectively). Conclusions: The most important cause of neoplastic changes in the cervix uteri is high-risk HPV infection, with evidence indicating that prolonged excessive smoking significantly exacerbates the persistence and progression of HPV infection, thereby influencing neoplastic changes in the cervix uteri. It is crucial for women to cease smoking in order to eradicate HPV infection from the body.

Oral human papillomavirus infection and genotyping in a cohort of people living with HIV.

This study is aimed at determining the prevalence of oral HPV infection and the risk indicators for oral HPV carriage in people living with HIV. Data on socio-demographics, sexual behavioral practices, and lifestyle practices of the participants were collected from 66 people living with HIV. The HIV parameters of each study participant were obtained from clinical records. Oral rinses obtained from each participant were subjected to HPV ELISA antigen test for screening and extracted DNA was subjected to nested PCR and whole-genome sequencing for genotyping. Approximately 36% (10 of 28) HIV-positive individuals had oral HPV carriage with one person carrying oncogenic type, HPV16. In addition, 80% (8 of 10) of those with HPV positivity by PCR are females, but with no statistically significant association. The CD4 count showed no significant association with oral HPV carriage in HIV positive individuals; however, age at first sex is a determinant of oral HPV infection in people living with HIV with positive association observed on both bivariate analysis and logistic regression (AOR: 150.49, 95% CI: 1.40-16,155.47, p = 0.036).

Impact of T Cell Exhaustion and Stroma Senescence on Tumor Cell Biology and Clinical Outcome of Head and Neck Squamous Cell Carcinomas.

Head and neck squamous cell carcinomas (HNSCC) have an overall poor prognosis, especially in locally advanced and metastatic stages. In most cases, multimodal therapeutic approaches are required and show only limited cure rates with a high risk of tumor recurrence. Anti-PD-1 antibody treatment was recently approved for recurrent and metastatic cases but to date, response rates remain lower than 25%. Therefore, the investigation of the immunological tumor microenvironment and the identification of novel immunotherapeutic targets in HNSCC is of paramount importance. In our study, we used tissue samples of n = 116 HNSCC patients for the immunohistochemical detection of the intratumoral and peritumoral expression of T cell exhaustion markers (PD-1, LAG-3, TIM-3) on tumor infiltration leukocytes (TIL), as well as the expression level of stromal senescence markers (IL-8, MMP-3) on tumor-associated fibroblasts. The clinical parameter of the vitamin D serum status as well as the histopathological HPV infection status of the tumor was correlated with the expression rates of the biomarkers and the overall patient survival. An increased peritumoral and intratumoral expression of the biomarkers PD-1 and TIM-3 significantly correlated with improved overall patient survival. A high peritumoral expression of LAG-3 correlated with better overall survival. A positive HPV tumor status correlated with a significantly elevated expression of PD-1 and TIM-3. Biomarkers of stromal senescence showed no influence on the patient outcome. However, the vitamin D serum status showed no influence on patient outcomes or biomarker expressions. Our study identified PD-1, LAG-3, and TIM-3 as promising targets of a therapeutic strategy targeting the tumor microenvironment in HNSCC, particularly among HPV-positive patients, where a higher expression of these checkpoints correlated with an improved overall survival. These findings support the potential of antibodies targeting these immune checkpoints to enhance treatment efficacy, especially in the context of bispecific targeting.

Upregulation of HPV16E1 and E7 expression and FOXO3a mRNA downregulation in high-grade cervical neoplasia.

Cervical cancer remains a significant global health concern, ranking as the fourth most prevalent cancer among women worldwide. Human papillomaviruses (HPV) transcribe many genes that might be responsible for cervical cancer development. This study aims to investigate the correlation between the expression of HPV16 early genes and the mRNA expression of human FOXO3a, a tumor suppressor gene, in association with various stages of cervical precancerous lesions. Eighty-five positive HPV16 DNA cervical swab samples were recruited and categorized based on cytology stages, i.e., negative for intraepithelial lesion or malignancy (NILM), atypical squamous cells of undetermined significance (ASC-US), low-grade squamous intraepithelial lesion (LSIL), atypical squamous cell cannot exclude HSIL (ASC-H), high-grade squamous intraepithelial lesion (HSIL). RT-qPCR was performed to amplify HPV16E1, E4, E6, E6*I, E7, and human FOXO3a mRNA expression in all samples. The relative expression of those genes was calculated using GAPDH as a control. Detection of FOXO3a mRNA expression in the cervical cancer cell line by RT-qPCR and meta-analysis of FOXO3a expression using the RNA-Seq dataset by GEPIA2 were analyzed to support the conclusions. Among the cervical samples, HPV16E1 and E7 were significantly increased expression correlating to disease severity. HPV16E4 mRNA expression was 100% detected in all LSIL samples, with a significant increase observed from normal to LSIL stages. Conversely, FOXO3a mRNA expression decreased with disease severity, and the lowest expression was observed in HSIL/squamous cell carcinoma (SCC) samples. In addition, similar results of FOXO3a downregulation were also found in the cervical cancer cell line and RNA-Seq dataset of cervical cancer samples. HPV16 early mRNA levels, including E1 and E7, increase during cancer progression, and downregulation of FOXO3a mRNA is a characteristic of cervical cancer cells and HSIL/SCC. Additionally, HPV16E4 mRNA expression was consistently detected in all LSIL samples, suggesting the presence of active viral replication. These findings might lead to further investigation into the interplay between HPV gene expression and host cell factors for targeted therapeutic strategies in cervical cancer management.

Distribution of HPV Types in Cervical Cancer in Pakistan: Implications for Screening and Vaccination Programs.

HPV plays a key role in the development of cervical cancer. This study aims to investigate the prevalence of HPV genotypes in patients with Squamous cell carcinoma (SSC) and Adenocarcinoma (ADC) at NORI cancer Hospital Pakistan, with the aim of improving screening and prevention strategies. Cervical scrapings were collected from 129 diagnosed cervical cancer patients. HPV typing was performed using a real-time PCR assay and sequencing. Among the patients, 73.6% (90/129) were HPV positive. Proportion of HPV positivity was observed within each group. The highest incidence of HPV was observed in the 50-60 years age group (80.9%), followed by the 40-to-50-year group (75.8%). The positivity rate declined in the 60-to-70-year-old (63.6%) and further in the 70-80 years (62.5%). Eight different HPV subtypes were identified, with HPV 16 being the most prevalent (80.0%), followed by HPV 18 (9.5%), HPV 45 in 2 (2.1%), HPV 31 in 1 (1.1%), HPV 35 in 1 (1.1%), HPV 59 in 1 (1.1%), HPV 66 in 1 (1.1%), and HPV 89 in 1 (1.15). Histologically, 89.2% of the cases were Squamous cell carcinoma (SCC) and 10.8% were Adenocarcinomas. In SCC patients, HPV 16 was found in 80.9% of cases, while in Adenocarcinoma patients, HPV 16 was detected in 66.7% of cases. The prevalence of high-risk HPV types, both vaccine and non-vaccine, targeted in Pakistan highlights the urgent need for widespread screening and vaccination programs. Tailored public health strategies are essential to effectively reduce cervical cancer rates and mortality.

Correlation of HPV-16, HPV-18 Genotypes with p16 Expression in Head and Neck Cancer: A Study from Western India

Objectives This retrospective study aims to elucidate the clinical associations of HPV-16, HPV-18, and p16 expression with clinicopathological parameters, risk behaviors, and survival outcomes in head and neck cancer patients from the western Indian population. Methods Clinical data of total 92 enrolled HNC patients diagnosed between years 2021 and 2023 were retrospectively collected from medical records. Formalin-fixed, paraffin-embedded blocks of all enrolled patients were collected whose p16 expression by immunohistochemistry tests were already performed. HPV-16 and HPV-18 infection was studied by real-time polymerase chain reaction. Associations between viral status, p16 expression, clinicopathological parameters, risk behaviors, and survival outcomes were assessed using SPSS statistical software version 20. p-Value ≤0.05 was considered to be statistically significant. Results Among the 92 enrolled HNC patients, HPV-16 infection was detected in only 12 (13%) patients, with the remaining 80 (87%) testing negative. No HPV-18 infections were observed in any patient. Additionally, p16 expression was positive in only 13 (14%) patients, while 79 (86%) showed negative expression. A statistically significant correlation was found between metastasis involvement and positive HPV-16 infection (p < 0.001), with all HPV-16-positive cases exhibiting metastasis. A trend was also noted between the base of tongue and other clinical site subtypes with positive HPV-16 infection (p = 0.063). However, no other clinicopathological or risk behaviors showed significant associations with HPV-16 infection and p16 expression. Overall survival analysis revealed that neither HPV-16 infection nor p16 expression served as significant prognosticators in the HNC patient cohort (p > 0.05). Conclusion This study provides comprehensive insights into the clinical relevance of HPV-16, HPV-18 infections, and p16 expression in HNC among the western Indian population. Understanding the associations between HPV-16, HPV-18, and p16 expression with clinicopathological parameters and survival outcomes may aid in optimizing patient management strategies, including personalized treatment approaches and targeted interventions. Further prospective studies are warranted to validate these findings and explore potential therapeutic implications.

Impact of knowledge of HPV positivity on cervical cytology performance in latin america.

Cervical cytology is recommended by WHO as a triage option in HPV-based cervical cancer screening programmes. We assessed the performance of cytology to detect CIN3+ without and with knowledge of HPV positivity. Women were screened with cytology and HPV across ESTAMPA study centres in Latin America. Screen-positives were referred to colposcopy with biopsy and treatment as needed. Cytology was initially interpreted without knowing HPV results. A subset of cytologies from HPV-positive women were re-interpreted at the same laboratories, with knowledge of HPV status, blinded to previous cytology and histological diagnosis. Performance indicators for cytology to detect CIN3+ without and with knowledge of HPV positivity were estimated. A total of 4,087 women were included, of which 490 had histologically confirmed CIN3 + (455 CIN3 and 35 cancers). Cytology sensitivity without knowledge of HPV positivity for CIN3+ was 47.2% (95% CI: 42.5-51.9), whereas with knowledge of HPV positivity, the sensitivity was higher (58.9%, 95% CI: 54.2-63.5), p < .0001. The specificity without knowledge of HPV was 89.4% (95% CI: 88.2-90.5), while with knowledge of HPV positivity was 78.9% (95% CI: 77.4-80.4), p < .0001. Performance estimates varied by study centre for cytology without knowing the HPV positivity, (range from 32.8% to 61.5% for sensitivity; range 80.7% to 98.6% for specificity). Similarly, performance varied with knowledge of HPV positivity (36.1% to 93.4% for sensitivity; 39.6% to 98.6% for specificity). The increase in sensitivity of cytology with HPV knowledge was limited and highly variable, reinforcing the need for alternative triage methods to support cervical cancer elimination goals.

Prediction of high-grade cervical precancerous abnormalities: The role of personal factors, vaginal microflora, sexually transmitted infections, and high-risk human papillomavirus.

High-risk human papillomavirus infection (HR-HPV) is necessary but not the only factor needed to develop cervical cancer. It is essential to estimate cervical cancer development risk in the population of high-risk HPV-positive women and to avoid unnecessary examinations and treatment in low-risk individuals. The study aimed to identify associations between different personal factors, vaginal microflora, sexually transmitted, high-risk HPV infection, and various degrees of cervical precancerous lesions. A study was performed in 2016-2020. The study group consisted of 112 patients with abnormal cervical cytology results referred for colposcopic examination. 120 women who came for a routine gynecological check-up were included in the control group. Material from the cervix and upper vaginal fornix was taken for pH measurement, wet mount microscopy, testing the six most common high-risk HPV DNA types (16/18, 31, 33, 45, 58), HPV E6/E7 mRNA, and 7 genital infections-C. trachomatis, N. gonorrhea, T. vaginalis, M. hominis, M. genitalium, U. urealyticum, U. parvum. Results showed that women with all grades of cervical intraepithelial neoplasia (CIN) more often were smokers, had increased vaginal pH levels, and had positive HR-HPV DNA and HR HPV E6/E7 mRNA expression. Abnormal vaginal microflora, especially types associated with aerobic vaginitis, and M. hominis were significantly more often found in women with CIN2+. The presence of C.trachomatis, U. parvum, and U.urealyticum did not differ between the groups. The most important factors independently associated with CIN2+ were positive high-risk HPV E6/E7 mRNA expression (OR 59.4, 95% CI 14.84-237.51), and positive high-risk HPV DNA (OR 3.9, 95% CI 1.16-13.23). Higher education level was associated with reduced risk of CIN2+ (OR 0.2, 95% CI 0.07-0.71). In conclusion, this study reports HR-HPV DNA of the most common six types and E6/E7 mRNA positivity as the most significant factors associated with CIN2+ lesions and higher education related to lower risk of high-grade cervical lesions.

Challenges and Considerations in Diagnosing and Managing p16+-Related Oropharyngeal Squamous Cell Carcinoma (OPSCC) with Neck Metastasis: Implications of p16 Positivity, Tobacco Exposure, and De-Escalation Strategies.

The incidence of patients showing neck metastasis and no obvious primary tumor at the initial diagnostic evaluation or neck cancer of unknown primary (NCUP) is rising. It is estimated that a relevant part of these tumors arises in the tonsillar crypts or base of the tongue and are p16+-related. However, today, the detection rate of the primary tumor is suboptimal. Identifying the primary tumor and its biomolecular characterization is essential since it influences the treatment administered, possibly reducing radiation fields and providing de-escalation to primary surgical management. However, p16 IHC (immunohistochemistry) might not be sufficient to diagnose HPV-related OPSCC. The other subset of patients discussed are the HPV-positive patients who have a history of tobacco exposure and/or p53 mutations. Possible factors that could negatively influence the outcomes of these patients are investigated and discussed below. So, this paper aims to analyze the diagnostic, bio-molecular, clinico-radiological, morphological, prognostic and therapeutical aspects of p16-positive OPSCC, highlighting the possible bias that can occur during the diagnostic and prognostic process. A narrative review was conducted to investigate the biases in the diagnostic and therapeutic process of two groups of patients: those who are p16-positive but HPV-negative patients, and those who are p16-positive and HPV-positive with exposure to traditional risk factors and/or p53 mutations. The keywords used for the literature research included the following: NCUP, OPSCC, p16IHC, HPV testing, p16 positive HPV negative OPSCC, p16 positive HPV positive OPSCC, tonsillectomy, tobacco exposure, p53 mutations, cystic neck metastasis, extranodal extension (ENE), radiotherapy, de-escalation and neck neck dissection. HPV-positive OPSCC has specific clinico-radiological features. Bilateral tonsillectomy should be considered for the identification of the primary tumor. P16 IHC alone is not sufficient for diagnosing HPV-related OPSCC; additional detection methods are required. The role of tobacco exposure and p53 mutations should be investigated especially in cases of HPV-positive tumors. Extranodal extension (ENE) must be taken into consideration in the prognostic staging of HPV-positive tumors. Surgical primary treatment involving neck dissection (ND) and bilateral tonsillectomy followed by adjuvant radiation may represent the most appropriate approach for N3 cases. Diagnosis, prognosis and therapeutical implications must be addressed considering clinical, biomolecular and morphological aspects. At least today, the numerous biases that are still present influencing the diagnostic and prognostic process do not permit considering de-escalation protocols. A precise and accurate diagnosis is required in order to adequately stage and manage p16+ OPSCC, particularly with neck metastasis. The role of tobacco exposure and/or p53 mutations must be considered not only in p16+ OPSCC but especially in HPV-positive OPSCC. Until a more accurate diagnosis is possible, ENE should be considered even in p16+HPV+ OPSCC. Primary surgery with unilateral ND and bilateral tonsillectomy might be the treatment of choice given the numerous diagnostic and prognostic pitfalls. Therefore, it is inappropriate and risky to propose de-escalation protocols in routine clinical practice due to the risk of undertreatment.

Epidemiological and typing features of HPV co-infections in MSM with mpox: A hospital-based prospective study

ObjectivesRecent mpox outbreaks highlight diverse transmission modes, with sexual behavior prominent in China's IIb strain. Nevertheless, despite HPV being a common sexually transmitted pathogen, there is a paucity of research into its coexistence and genotype distribution within this patients population. MethodsWe conducted a study at the Third People's Hospital of Shenzhen from May to September 2023. We collected information on mpox patients, including their sociodemographic and clinical characteristics. Anal swabs were collected for HPV genotyping. ResultsAmong 73 MSM mpox cases, HIV positivity was 56.2 % (41/73) and HPV positivity 80.4 % (41/51). Multiple HPV infections were prevalent (56.9 %, 29/51), especially among HIV-positive (93 %, 27/29). HPV16 (29.3 %, 12/41) was the most common high-risk genotype, followed by HPV59 (26.8 %, 10/41), HPV42 (19.5 %, 8/41), HPV6 (14.6 %, 6/41), and HPV54 (14.6 %, 6/41). HPV infection was significantly associated with HIV infection (p = 0.001). Additionally, HIV infection (p = 0.001) and PCR-detected positive sites for mpox (p = 0.047) were associated with high-risk HPV infection. ConclusionsIn this study, most of the mpox patients were found to be infected with HPV, mostly with high-risk types and multiple infections. Therefore, it is strongly recommended that mpox-infected individuals intensify HPV-related screening, prevention, and treatment measures.

Histomorphology based prediction of p16 immunopositivity and p16/HPV DNA co-positivity in oral squamous cell carcinoma

The histomorphological features predictive of p16 and human papilloma virus (HPV DNA) positivity in oropharyngeal carcinoma have been a matter of much debate. However, only few studies have been done on oral squamous cell carcinoma (OSCC) to correlate the histomorphological features with p16 and HPV DNA positivity. Oral squamous cell carcinoma has distinct etiopathogenesis, treatment and prognosis as compared to oropharyngeal carcinomas.A total of 800 oral squamous cell carcinoma biopsy cases were evaluated for features suggestive of HPV infection like basaloid appearance, absence of stromal reaction, nests and lobules of tumor cells with pushing borders, central necrosis, lympho-epithelial morphology, koilocytes, and non-keratinizing or hybrid morphology. Immunohistochemistry was performed for p16 expression (E6H4 clone, CINtec histology, Roche diagnostics). The cases which showed 2+/3+ (from moderate to high intensity) staining with >75 % cells were considered as p16 immunopositive. All the p16 immunopositive cases were subjected to real-time PCR (polymerase chain reaction) for HPV DNA detection to confirm HPV positivity. A total of 139 (17.37 %) OSCC cases were p16 immunopositive and out of these 104 (104/139, 74.8 %) cases showed HPV-DNA PCR positivity. None of the features were found to be predictive on multivariate logistic regression analysis. However, on bivariable analysis, nest/lobule with pushing border was the only histopathological feature which had a significant correlation with p16 immunopositivity (P value = 0.0001) and p16 and HPV DNA copositivity (P value = 0.0001). (Fisher's exact test -two tailed).To conclude-morphology is not really predictive of HPV positivity in OSCC cases. Only one feature- nests and lobule with pushing border is suggestive on bivariable analysis.

Comparative study of triage strategies for women with atypical squamous cells of undetermined significance in the post-vaccine era.

The research focused on a comparative analysis of triage strategies for women with Atypical Squamous Cells of Undetermined Significance (ASC-US) before and after receiving the HPV vaccine, aiming to optimize cervical cancer prevention strategies, especially in resource-limited healthcare settings. Between September 2018 and December 2023, 7,511 women aged 21 years or older who underwent liquid-based cytology for cervical cancer screening were recruited. Women diagnosed with ASC-US were included in the study. All participants underwent HPV testing and liquid-based cytology examination, and those with abnormal results were referred for colposcopy. Women with abnormal colposcopy findings underwent further histopathological examination. The gold standard for diagnosis was pathological, with cervical intraepithelial neoplasia grade 2 or higher (CIN2+) on histology as the endpoints. In the final analysis, 933 women with ASC-US were enrolled as the unvaccinated group, with 179 of them testing positive for HPV 16/18. Assuming that all women would receive the bivalent vaccine targeting HPV 16/18 in the post-vaccine era, and given that the vaccine protection rate is 100% against HPV 16/18, then 754 women excluding those of HPV 16/18 positive would comprise the vaccinated group. In the unvaccinated group, the overall HPV positivity rate was 59.27% among ASC-US women, with a 100% HPV prevalence rate among those with CIN2+ lesions. The combination genotyping model of HPV16/18 showed the highest specificity (81.77%) and the lowest referral rate (32.37%). In the vaccinated group, the HPV positivity rate was 49.61% among ASC-US women, with a 100% HPV prevalence rate among those with CIN2+ lesions. The specificity of HPV33/58 was the highest (86.99%), and the colposcopy referral rate was lowest (27.54%), with statistical significance. Sensitivity, positive predictive value, and negative predictive value were not statistically significant. HPV16/18 demonstrated a more efficacious triaging effect in the unvaccinated group. HPV33/58 will potentially replace HPV16/18 as the priority screening genotyping among vaccinated populations.

Abstract C017: Race-Specific Patterns of Vaginal Microbiome and HPV Infections Predict Risk of Precancerous Cervical Lesions

Abstract Background: Our previous research indicated that high-grade squamous intraepithelial lesions (HSIL), precursors to cervical cancer (CCa), are influenced by the vaginal microbiome (VMB) differentially by self-reported race. VMB enriched for certain anti-inflammatory Lactobacillus species confer protection against the risk of HSIL for white, but not Black women. Black women exhibit greater VMB diversity (associated with HPV tumorigenesis) and a lower prevalence of anti-inflammatory Lactobacillus species. But not all Black women with diverse VMB develop HSIL. Objective: Given HPV's essential role in cervical cancer, we aimed to assess the influence of sociodemographic, clinical, and microecological HPV attributes on HSIL risk within the context of the vaginal microbiome's variation by race. Methods: The VMB and HPV profiles of 1,168 women were analyzed alongside over 100 sociodemographic and clinical variables. Spearman’s correlation, Point-Biserial Correlation, chi-squared tests, and machine learning models, were employed to evaluate associations between sociodemographic, clinical, and microecological variables with HSIL, with a particular focus on the impact of race on these relationships. Results: The cohort was predominantly Black (72%), showing no significant race-based differences in HPV positivity (45%). However, HSIL incidence was nearly twice as high in Black (7%) compared to White women (4%). Women with HSIL had a higher prevalence of non-Lactobacillus-dominant VMB (68% vs. 51%) and were more likely to be HPV+ (73% vs. 44%) than those without HSIL. Co-infection with multiple HPV strains was associated with a higher risk of CIN3. Specifically, HPV16, HPV56, HPV58, HPV33, and HPV42 infections were significantly associated with higher CIN3 risks. The random forest model identified the relative abundance of Lactobacillus crispatus as the most essential factor in predicting CIN3 risk, followed by the number of HPV strains per sample. However, differential abundance analysis specifically indicated lower L. crispatus levels were associated with HSIL in white but not Black women. Conversely, HPV abundance was only associated with HSIL in Black women. Additionally, Black patients with 2 or more HPV co-infections had a higher likelihood of being diagnosed with CIN3+ compared to white counterparts. ML models highlighted different HPV-specific HSIL predictors by race: HPV number, and HPV 56, 16, and 33 in Blacks; sexual partners, HPV 90, 56, and 16 in whites. Conclusions: Our findings reveal race-specific interplays among individual risk factors, HPV infection, and VMB composition in HSIL development. The identification of key microbial and viral predictors highlights the importance of personalized approaches in preventing and managing HSIL, particularly among racially diverse populations. Citation Format: Katherine Y. Tossas, Bin Zhu, Katarzyna Tyc, Myrna Serrano, Jerome Strauss, Gregory Buck. Race-Specific Patterns of Vaginal Microbiome and HPV Infections Predict Risk of Precancerous Cervical Lesions [abstract]. In: Proceedings of the 17th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2024 Sep 21-24; Los Angeles, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2024;33(9 Suppl):Abstract nr C017.

Oral Mucosal Human Papillomavirus and Epstein-Bar Virus Rates in Patients with Dry Mouth and/or Sjögren's Syndrome in a Hungarian Cohort.

To find an association between oral mucosal human papilloma- and/or Epstein-Barr (HPV, EBV) virus infection in patients with dry mouth and/or Sjögren's syndrome (SS) compared to healthy controls and to find connections with salivary gland histopathological alterations. Ninety-two participants were divided into four groups: 1. healthy controls (n &#61; 32); 2. xerostomia (n &#61; 28); 3. hyposalivation (n &#61; 22); and 4. SS groups (n &#61; 10). To detect virus infection brush biopsy was outlined in all groups. Detections of virus-specific sequences were achieved with polymerase chain reaction (PCR). Lip biopsy and histopathological assessment was performed in groups 2, 3 and 4. HPV positivity of oral mucosal cells was shown in group 1: 1 (3.12%); group 2: 3 (10.7%); group 3: 2 (8.26%); and in group 4: 0 of the samples. EBV was present in group 1: 14 (43.7%); group 2: 17 (60.7%); group 3: 6 (27.3%); and in group 4: 5 (50%) of the cases. There was no statistically significant difference between the attributes. Intact salivary gland in 28.2%, chronic sialadenitis in 28.2%, stromal fibrosis in 6.5%, lipomatous atrophy in 8.6%, fibrous atrophy in 6.5% and positive focus score (SS) in 26.1% were found in the subjects. Neither HPV nor EBV infection caused statistically significantly more histological abnormalities. Orofacial mucosal HPV and/or EBV DNA rates did not differ statistically significantly in patients with xerostomia or hyposalivation or SS compared to healthy controls, therefore, it cannot prove the provocative role of these viruses in dry mouth and/or SS. Neither dry mouth nor SS were accompanied by statistically significantly more salivary gland alterations in HPV- and/or EBV-positive subjects; these alterations are frequent in the virus-negative cases too.

Expression and prognosis of DSG-2, CXADR, CD46 in head and neck squamous cell carcinoma

ObjectivesInvestigating the expression and prognostic significance of adenovirus receptors DSG-2, CXADR and CD46 in head and neck cancer. Methods104 patients with HNSCC (77 OPSCC, 27 LSCC) were retrospectively included in the study. Immunohistochemical staining was performed on all selected slides to detect the expression of DSG-2, CXADR, CD46 and the immunoreactive score (IRS) was determined from the number of positively stained tumor cells and their staining intensity. Furthermore, the respective HPV status was determined by immunohistochemical staining against p16 and HPV-PCR. Results81.7 % of the tumors showed DSG-2, 34.6 % of the tumors showed CXADR and 57.7 % of the tumors showed CD46 expression. A high DSG-2 IRS correlated significantly with an advanced tumor size (p= 0.003), increased grading (p=0.012) and positive HPV status (p=0.024) in OPSCC. A high CXADR IRS was significantly associated with a positive lymph node status (p= 0.041) in LSCC and an advanced AJCC stage (p= 0.012) and a positive HPV status (p= 0.009) in OPSCC. No significant correlation could be shown regarding CD46 expression and clinical tumor data. There was no effect of DSG-2, CXADR, and CD46 expression on 5-year overall and on 5-year disease-free survival. ConclusionNo prognostic significance of the expression of DSG-2, CXADR or CD46 in HNSCC was seen. DSG-2, CXADR and CD46 are expressed in HNSCC, so that optimization of oncotherapy with adenoviral vectors appears promising. Due to the significantly increased expression of DSG-2 and CXADR in advanced OPSCC tumors, there is potential for optimizing oncotherapy here in particular.

Anal high-grade intraepithelial neoplasia and cancer in women living with HIV and HIV-negative women with other risk factors.

To determine the prevalence and the risk factors for anal high-grade intraepithelial neoplasia and anal cancer (HSIL+) in women living with HIV (WLWHIV), and to compare them to HIV-negative women with other risk factors. Prospective cohort study. WLWHIV and HIV-negative women with other risk factors were included. Screening for anal HSIL+ using anal cytology and HPV testing was performed. A high-resolution anoscopy with directed biopsy was also performed in patients with an abnormal cytology result or a positive HPV testing for high-risk (HR) genotypes, and in those with anal symptoms. The period prevalence of anal HR-HPV infection and histological HSIL was 57.9% and 10.9% among WLWHIV, and 60.8% and 9.2% among HIV-negative women. The prevalence of anal HPV 18 infection was higher in WLWHIV. The risk factors for anal HSIL+ in WLWHIV included anal HPV 16, other HR genotypes and low-risk genotypes infection, as well as a history of vulvar HSIL+. In HIV-negative women, the risk factors included anal HPV 16 infection, history of anogenital warts and of vulvar HSIL+, and immunosuppressive treatment. A high prevalence of anal HPV infection and HSIL was observed in WLWHIV and women with other risk factors. Both groups share anal HPV 16 infection and history of vulvar HSIL+ as risk factors for the development of anal HSIL+. Genotyping for anal HPV 16 may help identify women at higher risk of anal cancer.

HPV integration: a precise biomarker for detection of residual/recurrent disease after treatment of CIN2-3

BackgroundThis study aimed to investigate whether persistent human papillomavirus integration at the same loci (PHISL) before and after treatment can predict recurrent/residual disease in women with CIN2-3.MethodsA total of 151 CIN2-3 women treated with conization between August 2020 and September 2021 were included. To investigate the precision of HPV integration, we further analyzed HPV integration-positive patients. Sensitivity, specificity, positive and negative predictive values (PPV and NPV, respectively), and the Youden index for predicting recurrence/residual disease were calculated.ResultsAmong the 151 enrolled CIN2-3 women, 56 were HPV integration-positive and 95 had HPV integration-negative results. Six (10.7%) experienced recurrence among 56 HPV integration-positive patients, which was more than those in HPV integration-negative patients (one patient, 1.1%). In the 56 HPV integration-positive patients, 12 had positive HPV results after treatment, seven had PHISL, and two had positive cone margin. Among the seven patients who tested with PHISL, six (85.7%) had residual/recurrent disease. PHISL was a prominent predictor of persistent/recurrent disease. The HPV test, the HPV integration test, and PHISL all had a sensitivity of 100% and a NPV of 100% for residual/recurrent disease. PHISL showed better specificity (98.0% vs. 82.0%, p = 0.005) and PPV (85.7% vs. 40.0%, p = 0.001) than the HPV test for predicting recurrence.ConclusionsThe HPV-integration-positive CIN2-3 women had much higher relapse rates than HPV-integration-negative CIN2-3 women. The findings indicate that PHISL derived from preoperative and postoperative HPV integration tests may be a precise biomarker for the identification of residual/recurrent CIN 2/3.