Human Papillomavirus Research Articles

Survivorship care plans and adherence to breast and cervical cancer screening guidelines among cancer survivors in a national sample

Abstract Purpose The impact of the components of survivorship care plans on adherence to cancer screening guidelines among cancer survivors is limited. We examined the association of receipt of treatment summaries, follow-up instructions, and type of doctor providing survivorship care with adherence to breast cancer screening (BCS) and cervical cancer screening (CCS) guidelines in female cancer survivors. Methods A cross-sectional analysis using Behavioral Risk Factor Surveillance System (BRFSS) data from 2014, 2016 and 2018 was conducted. BCS and CCS-eligible women were aged 40–74 and 30–64, respectively. BCS adherence was defined as a mammogram within 2 years and CCS adherence as a pap smear within 3 years or HPV test within 5 years. Univariate analysis with chi-square and multivariable logistic regression are reported. Results 5,001 BCS and 3,014 CCS-eligible survivors were identified. In the BCS group, recipients of treatment summaries and follow-up instructions were significantly more adherent with BCS (84.1% vs. 77.4%; 83.4% vs. 74%, respectively, p < 0.001). In the CCS group, recipients of follow-up instructions were significantly more adherent with CCS (78.1% vs. 67.7%, p < 0.001). In both groups, there was no significant difference in BCS or CCS based on type of physician providing care (p = 0.087). On multivariate analysis, receipt of follow-up instructions was the only factor significantly associated with BCS (OR:2.81; 95%CI:1.76–4.49) and CCS (OR:3.14; 95%CI:1.88–5.23). Conclusions Follow-up instructions, as part of survivorship care plans, have the strongest association with BCS and CCS among female cancer survivors. Additional research should focus on improving the distribution of survivorship care plans, particularly follow-up instructions, as a method to increase BCS and CCS among cancer survivors.

Risk of High-Grade Cervical Dysplasia in Older Patients With a Positive Human Papilloma Virus Cervical Screening Test

Introduction The Australian National Cervical Screening Program has mandated management algorithms that are uniform across all age groups, but evidence is emerging that perhaps the risk of high-grade squamous intraepithelial lesion (HSIL) may decrease in the postmenopausal period. Objective The aim of the study is to identify if patients ≥50 years of age referred to a tertiary colposcopy service have a different risk of HSIL or greater (+). Materials and Methods This is a retrospective cohort study of 3239 referrals to a hospital colposcopy clinic with a positive human papillomavirus (HPV) cervical screening test between December 2017 and May 2023. Patients were included if they were aged 25–74 years, had a cervix, and completed an outpatient colposcopic assessment. Medical records were analyzed to determine the incidence of HSIL+ for each age group and compared them using χ2 and multivariable regression. Results The overall incidence of invasive carcinoma was 0.93% and HSIL/adenocarcinoma-in-situ was 20.38%. Patients ≥50 years of age were less likely to have HSIL or greater diagnosed than those <30 (p < .0001) or those 30–49 (p < .0001), even after considering referral HPV subtype and reflex cytology (OR = 0.45, 95% CI = 0.34–0.59). The risk of invasive carcinoma in those aged ≥50 was no different than those aged 30–49 (p = .982). Conclusions We observed a lower incidence of HSIL or greater with increasing age even after considering referral HPV subtype and reflex cytology. As further data emerges regarding the risk of high-grade disease in those ≥50 years of age, refinement of the risk-based screening algorithm to account for advanced age may be required.

The Risk Factors of High-Risk Human Papillomavirus Susceptibility and Clinical Features in HIV-Positive Patients with Anal Condyloma Acuminatum: A Retrospective Cohort Study

The Risk Factors of High-Risk Human Papillomavirus Susceptibility and Clinical Features in HIV-Positive Patients with Anal Condyloma Acuminatum: A Retrospective Cohort Study

Human papillomavirus vaccine uptake among adolescent survivors of hematopoietic stem cell transplant

Human papillomavirus vaccine uptake among adolescent survivors of hematopoietic stem cell transplant

Human umbilical cord mesenchymal stem cells small extracellular vesicles-derived miR-370-3p inhibits cervical precancerous lesions by targeting DHCR24

Abstract Background Cervical cancer is often caused by persistent high-risk human papillomavirus (HPV) infection, causing precancerous lesions. Human umbilical cord mesenchymal stem cells-derived small extracellular vesicles (hucMSC-sEV) exhibit diverse effects on tumors. This study investigates hucMSC-sEV, the impact and mechanisms on HPV-positive cervical precancerous lesion cells to provide new treatment insights. Materials and Methods We previously obtained hucMSC and hucMSC-sEV. In vitro experiments evaluated hucMSC-sEV effects on the proliferation and migration of S12 cells (derived from cervical precancerous lesions). Bioinformatics identified key microRNA components, and their impact on S12 cell proliferation and migration was investigated. The target gene of the microRNA component was predicted and confirmed via bioinformatics and dual-luciferase reporter assays. Lentiviral systems overexpressed target gene in S12 cells to examine the effects on microRNA impacts. SH-42 inhibitor was used to investigate target gene treatment potential. Immunohistochemistry assessed target gene expression in cervical precancerous lesions tissue. Results hucMSC-sEV significantly inhibited S12 cell proliferation and migration. Bioinformatics identified miR-370-3p as an effective cargo, which also suppressed S12 cell proliferation and migration. miR-370-3p was confirmed targeting DHCR24 (24-Dehydrocholesterol Reductase). DHCR24 overexpression reversed miR-370-3p’s inhibitory effects, while SH-42 counteracted DHCR24 overexpression’s promoting effects. Clinical specimen analysis supported these findings, demonstrating a positive correlation between DHCR24 protein expression and cervical precancerous lesions’ progression. Conclusions hucMSC-sEV inhibits S12 cell proliferation and migration, mediated by miR-370-3p targeting DHCR24 to regulate cellular cholesterol content. DHCR24 inhibition reduces the cholesterol level and cell functions, suggesting its potential as a therapeutic target in cervical precancerous lesions.

Validity of Mehrviru® and Sacace® in molecular detection of HPV: a latent class analysis.

Valid diagnostic tests for human papillomavirus (HPV) detection are crucial to identify individuals at high risk of cervical cancer. We assessed and compared the validity of Mehrviru HPV genotyping and Sacace (HPV Genotypes 14 Real-TM Quant) for molecular detection of 14 high-risk human papillomaviruses. We used three HPV test results to identify HPV-positive individuals (14 high-risk genotypes) in a specialist gynecology clinic. The HPV test results were collected using Mehrviru®, Sacace®, and a third kit from the clinical diagnostic laboratory. We used Latent class analysis to determine the actual status of HPV infection in study participants. The sensitivity, specificity, positive and negative predictive values, positive and negative likelihood ratios, Youden, and area under the ROC curve indices of each diagnostic kit and their 95% confidence intervals were calculated. The agreement between the Mehrviru and Sacace kits was determined using the Kappa statistic. We examined 117 women at high risk of HPV infection. The mean age (SD) was 37.2 (9.1). According to LCA, 28.6% of participants had an HPV infection. The sensitivity and specificity (95% CI) of the Mehrviru were 90.8% (73.7-97.2%) and 90.9% (82.1-95.6%), and corresponding figures for Sacace were 92.0% (72.3-98.1%) and 97.4% (90.2-99.3%). The kappa index between the Mehrviru and Sacace kits was 69.7% (55.6-83.9%). The area under the ROC curve for Mehrviru and Sacace test were 91.4% (85.7-97.1%) and 94.4% (89.3-99.5%). There is an excellent agreement between Mehrviru and Sacace test results, and the diagnostic accuracy indices were similar.

Highly sensitive deep panel sequencing of 27 HPV genotypes in prostate cancer biopsies results in very low detection rates and indicates that HPV is not a major etiological driver of this malignancy.

Human papillomavirus (HPV) has been proposed to contribute to the carcinogenesis of prostate cancer. However, previous studies have yielded conflicting results. This study aims to add useful information to the ongoing discussion concerning the association between HPV infection and prostate cancer. We used two high-throughput next-generation sequencing (NGS) approaches to detect HPV RNA in malignant and adjacent normal (AN) prostate tissue (cohorts 1 and 2) and HPV DNA from carcinogenic and probably/possibly carcinogenic-classified HPV types (cohort 3) in malignant prostate, AN prostate, and benign prostatic hyperplasia (BPH) tissues. In total, 0% (cohort 1: 0/83, cohort 2: 0/16) of the malignant prostate tissue samples and 0% (cohort 1: 0/23, cohort 2: 0/8) of the AN prostate tissue samples were positive for HPV RNA. A total of 8.3% (1/12) of the BPH samples, 0% (0/28) of the AN samples, and 0.8% (1/132) of the malignant prostate samples were positive for HPV16 DNA. However, the normalized read count of the HPV16-positive malignant sample was close to the cut-off. In addition, no other carcinogenic-classified HPV types were detected in any of the BPH, AN, or malignant prostate tissue samples. Our study does not support HPV infection as a major contributor to the etiology of prostate cancer.

Prevalence, genotyping and phylogenetic analysis of HPV infecting Palestinian women.

Human Papillomavirus (HPV) is a significant global public health concern due to its association with cervical, other anogenital, and oropharyngeal cancers. This study aimed to evaluate the prevalence and the phylogenetic relationships of HPV among Palestinian in order to inform public health strategies. A cross-sectional study was conducted between September 2023 and April 2024 involving 379 Palestinian women over the age of 18 from 11 governorates in the West Bank. Cervical swabs were collected and analyzed using nested PCR and Sanger sequencing methods to detect and genotype HPV. The study also included phylogenetic analysis to understand the genetic relationships between HPV strains. The overall HPV prevalence was 14.5%. The highest prevalence was observed in the 20-29 age group (19.6%), the Middle region of the West Bank (19.0%), and lower educational attainment. Genotyping revealed a diverse distribution of HPV types, with HPV 11 and HPV 6 being the most common low-risk types, while HPV 16 was the most common high-risk type. About 21.8% of the detected strains were high-risk strains. Phylogenetic analysis indicated significant regional clustering of HPV strains. The study highlights the need for targeted public health interventions, including vaccination and regular screening, particularly for younger women and those with lower educational attainment. Continued surveillance and research are essential to reduce the burden of HPV-related diseases in the West Bank, Palestine.

OCCURRENCE OF HUMAN PAPILLOMAVIRUS (HPV) IN WOMEN OF LGBTQIA+ COMMUNITY TREATED IN MANAUS, AMAZONAS

Human papillomavirus (HPV) is a significant etiological factor for several types of cancer, including cervical carcinoma, with 80% of cases of cervical cancer occurring in Latin America. In Brazil, cervical carcinoma is the third most prevalent cancer among women, and the northern region and the state of Amazonas have the highest incidence in the country. However, not all women benefit from cervical cancer screening measures. Most women in the LGBTQIA+ community do not seek gynecological health services since they often face barriers related to female homosexuality when accesing sexual health services. Using PCR, this study perfomed the molecular detection and genotyping of HPV in thirty-six patients (20 homosexual and 16 bisexual women) from Manaus, Amazonas, Brazil. Samples of the cervical epithelium were taken during the preventive examination and genotyping was perfomed to detect HPV 16, 18, 31 and 58. Of the thirty-six patients analyzed in this study, 88% of bisexual women (n=14) and 60% of homosexual women (n=12) tested positive for HPV. The HPV 18 genotype was the most frequent among the findings. The results indicate that homosexual and bisexual women are more vulnerable to HPV infections due to the lack of more specific gynecological care directed to this group, which includes the lack of training of professionals to deal with the particularities of this population group. Therefore, more HPV screening studies for LGBTQIA+ women and improved training for specialized health professionals are needed.

HPV Vaccine Awareness and Uptake Among Sexually Transmitted Infections Clinic Users: A Cross-Sectional Study in Bologna, Italy

Human Papillomavirus (HPV) infection poses a significant health risk, particularly for high-risk groups such as men who have sex with men (MSM), people living with HIV (PLHIV), and transgender individuals. Despite the availability of effective vaccines, uptake among these groups remains suboptimal due to various social and behavioral barriers (BeSD). A cross-sectional survey was conducted at the Sexually Transmitted Infections (STIs) clinic in Bologna, Italy, from 8 April to 12 April 2024 using a paper questionnaire, investigating HPV vaccine uptake and BeSD factors influencing vaccination decisions. Statistical analyses included descriptive statistics and multivariate logistic regression. Among the 236 respondents, PLHIV and transgender individuals demonstrated lower uptake rates (60.0% and 15.6%) if compared to women under 30 years old (72.7%). Concern about HPV infection varied significantly across groups, with MSM showing the highest worry (48.7%). Perceptions of vaccine safety and access were mixed, influencing vaccination decisions. Multivariate analysis indicated that age inversely correlated with infection worry (OR: 0.94, 95% CI: 0.91–0.98), while being a woman under 30 (OR: 164.0, 95% CI: 17.2–1560.18) or MSM (OR: 3.53, 95% CI: 1.37–9.11) was positively associated with vaccine uptake. The study identifies disparities in HPV vaccine uptake among STI clinic users in Bologna, Italy, emphasizing the need for targeted public health campaigns. These campaigns could engage STI clinics and address awareness, safety perceptions, and access barriers to enhance vaccination coverage among sexual and gender minorities.

Pioneering molecular screening for cervical precursor lesions and cervical cancer in sera

Cervical cancer is a significant public health issue in Mexico and many developing countries. Early detection is crucial for combating this disease. The official screening test for cervical cancer is cytology, but this technique faces several barriers, including methodological, educational, and sociocultural challenges. Liquid-based cytology is an improved version of this test, however it does not address the aforementioned complications. Biomarkers for cervical precursor lesions and cervical cancer can improve timely detection of the disease. A previous study from our group identified four circulating human proteins as potential biomarkers for these conditions. For molecular screening, we selected GAPDH as the biomarker for cervical precursor lesions and HNRNPA1 as the biomarker for cervical cancer -chosen from the three previously identified options based on antibody availability- to be detected in sera. Participants underwent a comprehensive panel of tests, including liquid-based cytology, PCR detection of Human papillomavirus (HPV), colposcopy, and histopathology -when applicable-. The last two tests were used as references for determining sensitivity and specificity, with histopathology being the gold standard for cervical cancer diagnosis. All the participants successfully received colposcopies (n = 99) and only those women with visible or suspected cervical lesions/malignancies were biopsied (n = 62). A subset of randomly selected biopsies underwent p16INK4a immunohistochemistry (n = 36). This study compares the performance of liquid-based cytology with the molecular screening. With colposcopy as reference, liquid-based cytology showed 30% sensitivity and 96% specificity, while the molecular screening showed 90% sensitivity and 43% specificity. With histopathology as reference, liquid-based cytology showed 21% sensitivity and 93% specificity, while the molecular screening showed 85% sensitivity and 61% specificity. The molecular screening outperformed the liquid-based cytology in several areas, including detecting true-positive cases, reducing false-negative cases by 34.62%, application time, simplicity of result´s categories, and acceptance among participants. An ideal screening test requires high sensitivity, maintains moderate specificity, and minimizes false negatives. Our proposed screening test meets these criteria, making it an ideal complement -or alternative- for cervical cancer screening.

Abstract PR003: Preventing and treating HPV-related Cancer with mRNA Therapy Expressing A DC-targeting Antigen

Abstract Persistent human papillomavirus (HPV) infection is linked to several malignancies, highlighting the need for effective therapeutic vaccines. In this study, we tested a lipid nanoparticle-encapsulated mRNA vaccine expressing tHA-mE7-mE6. By mutating the E6 and E7 proteins to reduce their tumorigenicity and fusing them with a truncated influenza hemagglutinin protein (tHA) for better antigen uptake, we improved the vaccine’s efficacy. The tHA-mE7-mE6 mRNA vaccine showed superior results compared to mE7-mE6 mRNA, achieving complete tumor regression and preventing new tumors in an E6 and E7 positive model. It elicited a strong CD8+ T-cell response, with antigen-specific CD8+ T-cells found in the spleen, blood, and tumors. Additionally, the therapy increased DC and NK cell infiltration into tumors. Overall, this study demonstrates that the tHA-mE7-mE6 mRNA vaccine induces a potent anti-tumor immune response, showing promise for treating HPV-induced cancers and preventing recurrence. Citation Format: William Jia. Preventing and treating HPV-related Cancer with mRNA Therapy Expressing A DC-targeting Antigen [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: RNAs as Drivers, Targets, and Therapeutics in Cancer; 2024 Nov 14-17; Bellevue, Washington. Philadelphia (PA): AACR; Mol Cancer Ther 2024;23(11_Suppl):Abstract nr PR003.

Systematic review and meta-analysis of the association between naturally induced IgG, IgM and neutralising antibodies to HPV16 and newly detected cervical HPV16 infection outcomes

BackgroundIt is unclear whether recurrent cervical human papillomavirus type 16 (HPV16) infections can be prevented by naturally induced HPV16 antibodies in unvaccinated healthy women.MethodsWe systematically searched the literature for studies...

Abstract A019: Analytical and clinical validation of HPV-SEQ, an NGS-based liquid biopsy platform for detection and quantification of human papilloma virus circulating tumor DNA

Abstract Human papillomavirus (HPV) infection is the primary causative factor driving the rising incidence of oropharyngeal squamous cell carcinoma (OPSCC). High-risk HPV16 and HPV18 strains are responsible for the majority of HPV related OPSCC cases, with ∼90% and 3-5% respectively. As HPV positivity is associated with improved treatment response and prognosis, correct determination of HPV status is critical for risk stratification and treatment management. While diagnostic tumor biopsy remains a useful tool for HPV profiling, the drawbacks of this approach include pain and discomfort caused by its invasive nature, as well as inability to repeat multiple serial biopsies over time. Noninvasive detection of circulating tumor (ct)HPV-DNA in plasma has emerged as an attractive investigative strategy for early disease diagnosis, real-time response monitoring, and cancer surveillance. However, to date, there is no standard or routine liquid biopsy-based HPV testing for patients with OPSCC. The accuracy of HPV detection in plasma depends on the metrics of the specific method used. Comparison of the most common approaches for detection of ctHPV-DNA shows that next-generation sequencing (NGS)-based assays are superior to that of qPCR and ddPCR, especially in patients with low disease burden. Furthermore, as the majority of current liquid biopsy tests are designed to detect viral E6 and E7 genes, it may comprise a potential challenge for detection of patient-derived ctHPV-DNA among individuals treated with therapeutic vaccines expressing non-oncogenic E6/E7 constructs, actively evaluated in clinical trial settings. To this end, we have developed HPV-SEQ, a novel NGS method designed to achieve maximum sensitivity for HPV16/18 L1 gene in DNA libraries prepared from plasma of OPSCC patients. Here we describe the thorough analytical characterization and clinical validation studies of this lab-developed test conducted and certified under clinical laboratory improvement amendments regulations. HPV-SEQ assay were assessed using both contrived samples and biological specimens by measuring a set of performance attributes including limits of blank, limits of detection, limits of quantitation, accuracy and precision. The assay has demonstrated a robust quantitative detection across a wide ctHPV-DNA range, with low level of background signal (<0.05 copies/sample) in healthy donors and qualitative cut-off of >1.39 copies for both HPV strains (ensuring sufficient differentiation between negative and low positive samples), as well as outstanding sensitivity and specificity, approaching 100%. Given the rapid decrease in sequencing cost and effective integration of sequencing data in clinical care, NGS is likely to displace other HPV detection tests over the next few years. As such, our well-validated, ultra-sensitive, plasma-based HPV profiling method with optimal analytical performance may represent a significant advancement in risk stratification, treatment management, and surveillance for patients with OCSCC and other HPV- driven malignancies. Citation Format: Samaneh Eickelschulte, Anna Starus, David H Murray, Anja K Keyser, Oliver Schauer, Johannes Fredebohm, Frederick Jones, Alexander T Pearson, Everett E Vokes, Ari J Rosenberg, Nishant Agrawal, Evgeny Izumchenko. Analytical and clinical validation of HPV-SEQ, an NGS-based liquid biopsy platform for detection and quantification of human papilloma virus circulating tumor DNA [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr A019.

Human Papillomavirus (HPV) Genotypes in Mixed Squamous Cell Carcinoma of the Penis: A Study of 101 Tumors.

Squamous cell carcinomas with two or more coexisting clearly different histological subtypes of penile carcinomas are designated as mixed carcinomas in current classification models. They represent about 10% of all penile carcinomas. The aim of this study was to detect HPV genotypes in these unusual tumors. Tumors were selected from an international series of 1010 patients with penile carcinomas. Mixed carcinomas were grouped, according to WHO recommendations, as follows: 1. Carcinomas with warty/basaloid features mixed with HPV-independent carcinomas and 2. HPV-independent subtypes mixed with each other. HPV detection and p16INK4a immunostaining were performed. For HPV detection, whole tissue section-PCR analyses were performed by SPF10-DEIA-LiPA25 (version 1). As expected, HPV was detected more frequently in HPV-associated mixed carcinomas than in HPV-independent mixed carcinomas. Carcinomas with basaloid or warty features mixed with other SCC subtypes showed an HPV positivity rate of 46% (33 of 72 tumors) compared with 7% found in tumors with nonwarty/basaloid morphology (2 of 29 tumors). Eleven high-risk HPV genotypes were identified and the most common was HPV16 (65%) usually associated with basaloid morphology. p16INK4a immunostaining was positive in 76% of HPV-positive tumors. As in nonmixed carcinomas, although in lower proportion, a variable array of HPV genotypes was detected in mixed carcinomas. Apparently, the presence of a non-HPV component in an otherwise typical HPV-associated type tumor does adversely affect the prevalence of HPV positivity. Any amount of HPV-associated morphology superior to 20% in a mixed tumor is sufficient to classify them as HPV-associated, a WHO requirement.

Abstract A012: Analytical validation of a cfDNA-based fragmentomic profiling assay for the detection and quantification of 14 HPV types in cervical cancer

Abstract Purpose: Human papillomavirus (HPV) is the primary cause of cervical cancer, one of the most common cancers affecting women globally. Despite advances in prevention and screening, there remains a critical need for reliable, non-invasive assays to detect and monitor HPV-driven cervical cancers. Liquid biopsy offers a promising solution to meet this need. Here, we report the analytical validation of a plasma-based fragmentomic profiling assay designed to detect and characterize circulating tumor tissue modified viral (TTMV)-HPV DNA from 14 HPV types associated with cervical cancer. Methods: A high-complexity, laboratory-developed test was developed that employs 68 primers and 34 probes to target 14 HPV types via droplet digital PCR (ddPCR). A quantitative algorithm was applied to integrate fragment counts to generate a TTMV-HPV DNA Score, reflective of the quantity of circulating tumor-derived HPV DNA. Analytical validation was performed by determining the limit of blank (LOB), limit of quantitation (LOQ), and limit of detection (LOD) for each HPV type. To measure the limits of assay performance, engineered samples were used for reference interval studies. Synthesized target-specific DNA regions of HPV16, HPV18, HPV31, HPV33, HPV35, HPV39, HPV45, HPV51, HPV52, HPV56, HPV58, HPV59, HPV66, and HPV68 were diluted to varying concentrations to perform assay validation. Results: The assay demonstrated excellent specificity, with LOBs ranging from 0 to 0.07 copies/µL across the 14 HPV types. LODs ranged from 0.20 to 1.02 copies/µL, indicating a high level of sensitivity. The assay also showed high accuracy and precision, with coefficients of variation (CV) below 20% for all HPV types across both intra- and inter-assay precision studies. Regression analysis confirmed strong linearity (R2 > 0.99) across a wide range of analyte concentrations. Conclusion: These results demonstrate that TTMV-HPV DNA assay accurately and reproducibly detects circulating HPV DNA for 14 HPV types commonly associated with cervical cancer, offering a valuable tool for both diagnosis and ongoing surveillance in clinical settings. Future studies will present the clinical validation in order to establish the prognostic utility of the assay in predicting the likelihood of HPV-driven cervical cancer in patients. Citation Format: Sunil Kumar, Joshua Hutcheson, David Conway, Michael Horan, Syandan Chakraborty, Chloe Wiseman, Alicia Gunning, Evgeny Skyrypkin, Cassin Williams, Catherine Del Vecchio Fitz, Piyush B Gupta. Analytical validation of a cfDNA-based fragmentomic profiling assay for the detection and quantification of 14 HPV types in cervical cancer [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr A012.

Abstract B018: Plasma-based circulating tumor tissue modified viral (TTMV)-HPV DNA for detection of recurrent HPV-driven anal cancer

Abstract Purpose: Squamous cell carcinoma of the anus (SCCA) is primarily driven by human papillomavirus (HPV) infection. Up to 30% of patients experience recurrence, necessitating improved surveillance strategies. Liquid biopsies have shown promise as a tool in oncology. This study investigates the clinical performance of circulating tumor tissue modified viral (TTMV)- HPV DNA in the post-treatment surveillance of SCCA. Patients and methods: This central IRB-approved retrospective cohort study included 117 patients diagnosed with HPV-driven SCCA from seven U.S. centers who had ≥1 TTMV-HPV DNA tests (NavDx®, Naveris, Inc., Waltham, MA) performed between March 2020 and June 2024. TTMV-HPV DNA for HPV subtypes 16, 18, 31, 33, and 35 were analyzed using droplet digital PCR. The diagnostic accuracy of TTMV-HPV DNA was evaluated by comparing test results with clinical evidence of SCCA recurrence determined through anoscopy, radiology, and/or tissue biopsy. Recurrence is defined as either a positive biopsy or subsequent salvage therapy. Results: Patients were primarily female (73%) with a median age 63 years. Median follow-up after initial treatment was 19 months. Forty-seven patients (40%) had pretreatment TTMV-HPV DNA testing, with a baseline detectability of 85.1% (40/47, 95% CI: 74.9–95.3). Of these, 24 were re-tested within 3 months post-treatment and 18 had undetectable TTMV-HPV DNA (75%). Those patients whose scores became undetectable had significantly longer recurrence-free survival than those whose scores remained detectable (log-rank test, p=0.01). One hundred and four patients (89%) underwent post-treatment testing, of which 91 provided relevant data for the analysis. Of these 91 patients, 27 (29.7%) experienced recurrence. Post-treatment testing demonstrated a per- patient sensitivity of 85.7% (24/28, 95% CI: 72.8-98.7). Of four patients with a false negative test, only one had biopsy-confirmed recurrence, whereas the remaining recurrences were not biopsied prior to starting salvage treatment. The positive predictive value of a single positive test was 97.6% (40/41; 95% CI: 92.8–100.0). One patient remains in active follow-up. Of the true positive post-treatment tests, 58% (14/24) predicted recurrence before clinical detection, with a median lead time of 59 days (range: 10–536). There were 141 negative post-treatment tests across 66 patients. This yielded a per-test specificity and negative predictive value of 99.3% (136/137, 95% CI: 97.8-100.0) and 97.2% (136/140, 95% CI: 94.9-99.9), respectively. Indeterminate TTMV-HPV DNA tests were rare (5/268,1.9%). However, clinically indeterminate findings (CIFs) were common, with 141 post-treatment CIFs identified across 52/117 patients (44.4%). TTMV-HPV DNA testing was able to correctly predict recurrence status in 94.3% of CIF cases. Conclusion: TTMV-HPV DNA testing is a sensitive and specific surveillance method for detecting early recurrence of HPV-driven SCCA. Citation Format: Shane Lloyd, Rafi Kabarriti, James Jabalee, Tyler Slater, Corbin Jacobs, Sean Inocencio, Michael Rutenberg, Chance Matthiesen, Kasha Neff, Gene-Fu F Liu, Tiffany M Juarez, Catherine DV Fitz, Stanley Liauw. Plasma-based circulating tumor tissue modified viral (TTMV)-HPV DNA for detection of recurrent HPV-driven anal cancer [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr B018.

The radiological features of HPV-positive vs HPV-negative OPSCC at a South African hospital

Background: Studies have found that, at presentation, human papillomavirus (HPV)-positive oropharyngeal squamous cell carcinoma (OPSCC) has a less advanced primary tumour, more advanced lymph node spread and commonly has cystic metastatic lymph nodes in comparison to HPV-negative OPSCC.Objectives: To compare the radiological features of HPV-positive and HPV-negative OPSCC in South African patients.Method: A retrospective cross-sectional study was conducted at a large South African hospital. Eligibility required a histologically proven OPSCC between 2007 and 2023; a p16 antigen test and, if positive, a confirmatory HPV DNA PCR test and a baseline pre-treatment contrast enhanced neck CT scan. All eligible HPV-positive OPSCC patients and a random sample of eligible HPV-negative OPSCC patients were enrolled.Results: Twenty-one HPV-positive and 55 HPV-negative OPSCC patients were recruited. There was no statistically significant difference in the tumour epicentre location, local advancement (≥ T3 in 67% and 71%, respectively, p = 0.54), mean primary tumour size (41 mm vs. 39 mm, p = 0.73), lymph node spread (bilateral or more in 67% vs. 82%, p = 0.22) or morphologically cystic lymph nodes (10% and 4%, p = 0.61).Conclusion: There was no statistically significant difference in the CT imaging appearances of HPV-positive and HPV-negative OPSCC in the studied sample of South African patients.Contribution: This study documents the radiological features of OPSCC in a small South African sample population, where HPV-positive and HPV-negative OPSCC could not be distinguished on CT criteria and did not display the classic features described in the literature.

Implementation of a Screening Program for High-Grade Anal Dysplasia in High-Risk Patients in a Tertiary Cancer Center.

The incidence of anal squamous cell carcinoma (SCC) has been increasing over the last decades. Human papillomavirus (HPV) infection accounts for more than 90% of anal cancers, and HIV co-infection increases the risk of invasive cancer. Men who have sex with men (MSM) with HIV are the highest risk group for developing anal high-grade squamous intraepithelial lesions (aHSILs), which can be found in 45%-50% of these patients and are precursor lesions for invasive cancer. Anal cytology is an effective screening tool, but it lacks sensitivity. High-resolution anoscopy (HRA) is the gold standard procedure for diagnosis and treatment of aHSILs. Recent data suggest that early detection and treatment of aHSILs could prevent the development of invasive cancer in this population. The objective of the study was to describe the implementation of an office-based screening program for anal cancer prevention in a Comprehensive Cancer Center in Brazil. Training included participation in the International Anal Neoplasia Society (IANS) HRA course at UCSF Medical Center Mount Zion in San Francisco, CA, USA, by three colorectal cancer surgeons. In-person and hands-on training was provided by a specialist through the AIDS Malignancy Consortium (AMC) of the US NIH. Equipment purchased and provided by the AMC included a colposcope with a digital camera, a hands-free mouse pedal, and a photo documentation imaging software program that allows images to be recorded for documentation and training purposes. The program was implemented in 2022 after a delay of more than two years due to the COVID-19 pandemic. An average of 24 exams are performed monthly. Patients with HIV aged 35 years or older who are undergoing antiretroviral therapy were recruited from the metropolitan area of Rio de Janeiro and referred by primary care providers for screening. Patients diagnosed with aHSILs are scheduled for in-office ablative treatment in the clinic. From March 2022 to June 2024, 324 exams were performed, and aHSIL was found in 38.2% of 220 high-risk patients, including 45 of 129 MSMs (34.9%), 6 of 19 transgender women (31.6%), and 33 of 72 women living with HIV (45.8%). A total of 69 treatments for aHSIL were performed in 62 patients. Patients are followed on a regular basis and long-term results are awaited, including the effectiveness of local therapy for aHSIL. The screening and treatment program was successfully implemented in a tertiary comprehensive Cancer Center. Team training and external proctorship were decisive for the achievement of benchmark standards. The program aims to permanently provide screening for the prevention of anal cancer through the detection and treatment of aHSIL within the National Cancer Institute of Brazil for populations considered at-risk for anal cancer.

Abstract A065: Estimation of HPV16-induced cervical cancer cells in routine Thin-prep cytology samples of cancer patients

Abstract Persistent infection with human papillomavirus has been implicated in the etiology of cervical cancer and sustained expression of its E7 oncoprotein has been rendered suggestive of malignant transformation. However, there are limited E7 protein-based detection assays. This might be due to paucity of data on steady-state expression level of E7 protein and the number of HPV-induced cervical cancer cells in routine Thin-prep cytology samples. So, the objective of the present study was to identify and estimate HPV16-induced cervical cancer cells in cytology samples of cancer patients, with the goal to generate a quantitative immunological method in the cytological diagnosis of cervical lesions. We collected 25 Thin-prep cytology samples of cervical cancer patients (PAP test confirmed) from Obstetrics and Gynecology department, AIIMS, New Delhi. Samples were conditioned to enrich cellular components and were then quantified using hemocytometer for total cell count, PAP staining for cancerous cell count and immunofluorescence using HPV16 E7 antibody for HPV16-induced cancerous cell count. The method devised efficiently found an average number of 7 × 103 cells/ ml to be E7 positive, out of total 27 x 104 cells/ ml. Although PAP test detected 71 x 103 cells/ ml to be abnormal or cancerous, only 10% of them were found to be HPV16 transformed, suggesting HPV E7 as the primary marker for detecting transformed cells and thereby, identifying patients at-risk and prompting a better prognosis. This simple and efficient method deduced allows optimizing the number of HPV16-induced cervical cancer cells from cytology samples which directly correlates to the amount of HPV-16 major oncoprotein- E7 expression, and the disease severity. Citation Format: Srishty Raman, Pranay Tanwar, Neerja Bhatla, Subhash Chandra Yadav. Estimation of HPV16-induced cervical cancer cells in routine Thin-prep cytology samples of cancer patients [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr A065.