Human Papillomavirus Status Research Articles

Role of TORS as De-Escalation Strategy in HPV-Related Oropharyngeal Cancer, What We Need to Know.

Human papillomavirus (HPV)-related oropharyngeal squamous cell carcinoma (OPSCC) presents unique challenges and opportunities for treatment, particularly regarding de-escalation strategies to reduce treatment morbidity without compromising oncological outcomes. This paper examines the role of Transoral Robotic Surgery (TORS) as a de-escalation strategy in managing HPV-related OPSCC. We conducted a comprehensive literature review from January 2010 to June 2023, focusing on studies exploring TORS outcomes in patients with HPV-positive OPSCC. These findings highlight TORS's potential to reduce the need for adjuvant therapy, thereby minimizing treatment-related side effects while maintaining high rates of oncological control. TORS offers advantages such as precise tumor resection and the ability to obtain accurate pathological staging, which can guide the tailoring of adjuvant treatments. Some clinical trials provide evidence supporting the use of TORS in specific patient populations. The MC1273 trial demonstrated promising outcomes with lower doses of adjuvant radiotherapy (RT) following TORS, showing high locoregional tumor control rates and favorable survival outcomes with minimal side effects. ECOG 3311 evaluated upfront TORS followed by histopathologically directed adjuvant therapy, revealing good oncological and functional outcomes, particularly in intermediate-risk patients. The SIRS trial emphasized the benefits of upfront surgery with neck dissection followed by de-escalated RT in patients with favorable survival and excellent functional outcomes. At the same time, the PATHOS trial examined the impact of risk-adapted adjuvant treatment on functional outcomes and survival. The ongoing ADEPT trial investigates reduced-dose adjuvant RT, and the DART-HPV study aims to compare standard adjuvant chemoradiotherapy (CRT) with a reduced dose of adjuvant RT in HPV-positive OPSCC patients. These trials collectively underscore the potential of TORS in facilitating treatment de-escalation while maintaining favorable oncological and functional outcomes in selected patients with HPV-related OPSCC. The aim of this scoping review is to discuss the challenges of risk stratification, the importance of HPV status determination, and the implications of smoking on treatment outcomes. It also explores the evolving criteria for adjuvant therapy following TORS, focusing on reducing radiation dosage and volume without compromising treatment efficacy. In conclusion, TORS emerges as a viable upfront treatment option for carefully selected patients with HPV-positive OPSCC, offering a pathway toward treatment de-escalation. However, selecting the optimal candidate for TORS-based de-escalation strategies is crucial to fully leverage the benefits of treatment de-intensification.

Transcriptionally Active Human Papillomavirus Infection in a Minority of Esophageal Squamous Cell Carcinomas in North America.

The role of Human papillomavirus (HPV) infection in esophageal squamous cell carcinoma (ESCC) is a topic of ongoing debate. This study used two screening approaches to look for evidence of HPV infection in esophageal squamous cell carcinoma. We initially checked for HPV infection in a randomly selected group of 53 ESCC cases. We did not detect any tumors positive for high-risk HPV. However, during clinical practice, we identified an HPV-positive ESCC in the distal esophagus, which tested positive for HPV16. This index case was TP53 wild-type, as determined by next-generation DNA sequencing (NGS). Since TP53 mutations are rare in other HPV-driven cancers, we improved our screening method by limiting our screen to a subset of ESCC cases without TP53 mutations. A second screen of 95 ESCCs (from 93 patients) sequenced by NGS revealed an additional 7 ESCCs with TP53 wild-type status (7.3% of the total). Of the 7 cases, 2 cases were found to be high-risk HPV positive. Both patients also tested positive for circulating cell-free HPV DNA and had a complete response to neoadjuvant chemoradiation. The index patient had microscopic residual tumor following neoadjuvant therapy. The patient underwent adjuvant immunotherapy and remained disease free after 22 months of surveillance. This study affirms the transcriptionally active status of high-risk HPV in a minority of ESCC patients in North America.

Prognostic outcome of cervical laser ablation using a holmium yttrium-aluminum-garnet (Ho:YAG) laser for the treatment of cervical intraepithelial neoplasia: A single-center retrospective study

ObjectiveAlthough cervical conization is considered a standard treatment for cervical intraepithelial neoplasia (CIN) 2/3, laser ablation can compensate for the disadvantages of the former. CO2, semiconductor, and holmium yttrium-aluminum-garnet (Ho:YAG) lasers are applied in ablation, but no previous studies have shown the effectiveness of any of these techniques. Here, we retrospectively analyzed the application of the Ho:YAG laser in our hospital to verify its efficacy, and discussed the methods for optimal recurrence detection. MethodsWe evaluated the recurrence rates of the pathological condition in patients who underwent laser ablation with a Ho:YAG laser for CIN2/3 at our institution from June 2012 to November 2021. We defined the recurrence as histologically confirmed CIN2 or more advanced stage. Age, preoperative diagnosis, human papillomavirus (HPV) genotype, and postoperative high-risk HPV status were recorded to establish their association with recurrence rates. ResultsWe performed surgery in 607 patients and the 2-year recurrence rate after interventions was 5.6%. Five patients were diagnosed with invasive cancer at the time of recurrence. Older age significantly correlated with higher risk of recurrence, but preoperative CIN grade and preoperative HPV 16/18 status did not significantly affect it. The postoperative high-risk HPV test was 100% sensitive for detecting recurrence. ConclusionsLaser ablation with the Ho:YAG laser yields promising results. Together with postinterventional management, high-risk HPV test after laser ablation should be conducted after diagnostic conization.This study received the approval from the Ethics Committee of the NHO Tokyo Medical Center (Ethics Committee approval number: R22-067).

The Prognostic Value of Human Papillomavirus Status in Penile Cancer: Outcomes From a Norwegian Cohort Study

IntroductionPenile squamous cell carcinoma (PSCC) can develop from human papillomavirus (HPV) infection. This study investigates if the prognostic value of the TNM stage groups or the components tumor stage (pT), grade of differentiation (Grade), lymphovascular invasion (LVI), and nodular stage (pN) depend on HPV status. Also, whether the value of tumor parameters (pT, Grade, and LVI) for predicting node-positive disease depends on HPV status was investigated. Patients and MethodsStored tumor tissue from 226 patients treated for PSCC in Western Norway between 1973 and 2023 was investigated for HPV DNA. Histopathological variables were reevaluated according to the current TNM classification. Disease course was registered from hospital records. Inclusion of an interaction term between HPV and TNM stage groups in Cox regression enabled analysis of whether cancer-specific survival (CSS) of the stage groups depended on HPV status. This was also done separately for pT, Grade, LVI, and pN. Logistic regression with interaction terms between HPV and the tumor parameters were used to investigate if their predictive value depended on HPV status. ResultsHPV DNA was detected in 43% of the tumors. Stratified by HPV status, there was no significant interaction term in the Cox regression between HPV status and TNM stage groups (P = .74). Similar results were found for pT (P = .94), Grade (P = .08), LVI (P = .91) and pN (P = .77). Moreover, there were no significant interaction terms in the logistic regression between HPV status and the tumor parameters pT, Grade, and LVI (all P > .2). ConclusionsThis study found that prognosis of the TNM stage groups and the components pT, Grade, LVI, and pN were not modified by HPV in PSCC. The value of pT, Grade, and LVI for predicting lymph node-positive disease was not affected by HPV status.

Association of Human Papilloma Virus, Cytomegalovirus, and Epstein-Barr Virus with Breast Cancer in Jordanian Women.

Background and Objectives: The investigation of oncogenic viruses and their potential association with breast cancer (BC) remains an intriguing area of study. The current work aims to assess evidence of three specific viruses, human papillomavirus (HPV), cytomegalovirus (CMV), and Epstein-Barr virus (EBV) in BC samples and to explore their relationship with relevant clinicopathological variables. Materials and Methods: The analysis involved BC samples from 110 Jordanian female patients diagnosed with BC and breast tissue samples from 30 control patients with no evidence of breast malignancy, investigated using real-time PCR. The findings were then correlated with various clinico-pathological characteristics of BC. Results: HPV was detected in 27 (24.5%), CMV in 15 (13.6%), and EBV in 18 (16.4%) BC patients. None of the control samples was positive for HPV or CMV while EBV was detected in only one (3.3%) sample. While (HPV/EBV), (HPV/CMV), and (EBV/CMV) co-infections were reported in 1.8%, 2.7%, and 5.5%, respectively, coinfection with the three viruses (HPV/CMV/EBV) was not reported in our cohort. A statistically significant association was observed between HPV status and age (p = 0.047), and between clinical stage and CMV infection (p = 0.015). Conclusions: Our findings indicate the presence or co-presence of HPV, CMV, and EBV in the BC subpopulation, suggesting a potential role in its development and/or progression. Further investigation is required to elucidate the underlying mechanisms that account for the exact role of oncoviruses in breast carcinogenesis.

Human Papillomavirus Is Rare and Does Not Correlate with p16INK4A Expression in Non-Small-Cell Lung Cancer in a Jordanian Subpopulation.

Background and Objectives: Human papillomavirus (HPV) was previously investigated in lung cancer with wide inter-geographic discrepancies. p16INK4a has been used as a surrogate for detecting high-risk HPV (HR-HPV) in some cancer types. This study assessed the evidence of HPV in non-small-cell lung cancer (NSCLC) among Jordanian patients, investigated the expression of p16INK4a, and evaluated its prognostic value and association with HPV status. Materials and Methods: The archived samples of 100 patients were used. HPV DNA detection was performed by real-time polymerase chain reaction (RT-PCR). p16INK4a expression was assessed by immunohistochemistry (IHC). The Eighth American Joint Committee on Cancer protocol (AJCC) of head and neck cancer criteria were applied to evaluate p16INK4a positivity considering a moderate/strong nuclear/cytoplasmic expression intensity with a distribution in ≥75% of cells as positive. Results: HPV DNA was detected in 5% of NSCLC cases. Three positive cases showed HR-HPV subtypes (16, 18, 52), and two cases showed the probable HR-HPV 26 subtype. p16INK4a expression was positive in 20 (20%) NSCLC cases. None of the HPV-positive tumors were positive for p16INK4a expression. A statistically significant association was identified between p16INK4a expression and the pathological stage (p = 0.029) but not with other variables. No survival impact of p16INK4a expression was detected in NSCLC cases as a group; however, it showed a statistically significant association with overall survival (OS) in squamous cell carcinoma (SqCC) cases (p = 0.033). Conclusions: This is the first study to assess HPV and p16INK4a expression in a Jordanian population. HPV positivity is rare in NSCLC among a Jordanian subpopulation. P16 INK4a reliability as a surrogate marker for HPV infection in lung cancer must be revisited.

P16 and HPV status in head and neck sarcomas and sarcomatoid carcinomas

Human papillomavirus (HPV)-positive oropharyngeal carcinoma is a distinct type of head and neck carcinoma with improved prognosis. p16 immunostaining is often used as a surrogate marker for HPV infection in this particular setting. The aim of this study is to estimate the prevalence of p16 staining and HPV infection in head and neck sarcomatoid carcinomas as well as head and neck sarcomas. 21 sarcomatoid carcinomas and 28 head and neck sarcomas were tested for p16 positivity using immunohistochemical staining, and for high-risk HPV infection using In situ hybridization (ISH). 24 % of sarcomatoid carcinomas and 21 % of sarcomas were positive for p16 staining. All 49 cases were negative for HPV ISH. The results confirm that p16 staining is not specific and may not be associated with HPV infection in non-oropharyngeal head and neck sites. They also indicate that non-oropharyngeal head and neck sarcomatoid carcinomas are not likely to be HPV related.

Histologic and Genomic Analysis of Conjunctival SCC in African and American Cohorts Reveal UV Light and HPV Signatures and High Tumor Mutation Burden.

Conjunctival squamous cell carcinoma (conjSCC) is more prevalent and aggressive in sub-Saharan African countries compared with the rest of the world. This study aims to compare the genomic, immunophenotypic, and histologic features between patients from the United States and Ethiopia, to identify etiopathogenic mechanisms and unveil potential treatment strategies. We compared histologic features and mutational profiles using whole exome sequencing, high-risk human papillomavirus (HPV) status, PD-L1 expression, and tumor-infiltrating lymphocytes in conjSCC tumors of patients from Ethiopia (ETH; n = 25) and the United States (from MD Anderson [the MDA cohort]; n = 29). Genomic alterations were compared with SCCs from other anatomic sites using data from The Cancer Genome Atlas. Solar elastosis was seen in 78% of ETH and 10% of MDA samples. Thicker tumors had higher density of CD8+ and CD3+ cells. HPV status was similar between the cohorts (ETH = 21% and MDA = 28%). The mean tumor mutation burden (TMB) was significantly higher in conjSCC (3.01/Mb, log10) and cutaneous SCC compared other SCC subtypes. ETH samples had higher TMB compared to the MDA cohort (3.34 vs. 2.73). Mutations in genes associated with ultraviolet light (UV) signature were most frequently encountered (SBS7b = 74% and SBS7a = 72%), with higher prevalence in the ETH cohort, whereas SBS2 and SBS13 signatures were more common among MDA HPV+ conjSCCs. Our findings suggest that UV exposure may play a major role in conjSCC, with a higher prevalence in the ETH cohort compared with the MDA cohort, where HPV also contributes.

Abstract LB137: Comparison of HPV virus-induced oropharyngeal cancer by self-reported race and the potential implications on treatment

Abstract African Americans exhibit lower survival rates from Human Papillomavirus (HPV)-induced squamous cell carcinomas in comparison to other population groups. While socioeconomic status and access to healthcare may contribute to these disparities, mounting evidence indicates that significant biological differences between diverse ancestries play a role in the severity of different cancers such as breast cancer, colorectal cancer, and squamous cell carcinomas. The purpose of the study is to investigate diagnostic and biological differences in HPV-induced squamous cell carcinomas of the oropharynx by self-reported race. The study population consists of oropharynx cases identified at the University of Michigan (non-Hispanic White) and through the Louisiana Tumor Registry (African American). A total of 52 individuals from two study sites were included in this study, 40 of whom are men, two are women and 10 are unknown. Forty-four percent (23/52) of individuals were African Americans (AA), and 46 % (19/52) were Non-Hispanic White (NHW). Formalin-fixed paraffin-embedded (FFPE) tumors from these patients were tested for p16 expression via immunohistochemistry as a proxy for HPV status. Fifty-two percent (27/52) tested negative (p16-) and 48% (25/52) tested positive (p16+). Total RNA from these samples was extracted, sequenced, and bioinformatically analyzed to test for the presence of HPV. Among AA samples, 61% were negative for both p16 and HPV RNA (p16-/RNA-), 26% were p16+/RNA-, 13% were p16+/RNA+ and none were p16-/RNA+. Among NHWs, 37% were p16-/RNA-, 16% were p16-/RNA+, 47% were p16+/RNA+ and none were p16+/RNA-. Interestingly, 6 of 20 HPV RNA- AA samples were p16+, whereas 0 of 7 HPV RNA- NHW samples were p16+. We performed data dimensionality reduction using Principal Component Analysis (PCA) implemented in the PCAtools Bioconductor package. Four groups were separated in a plot of the first two PCs, comprising, (i) all p16+/RNA+ (18/18), (ii) dominantly p16-/RNA- (6/7), (iii) p16-/RNA- mixed with p16+/RNA-, and (iv) p16-/RNA+ in the middle of all the groups and mixing with some p16+/RNA+ samples. A total of 13/17 (76%) p16-/RNA- samples in group (iii) that were mixed with p16+/RNA- samples were AA, 3 (18%) were NHW and 1 was unknown. Based on clinical data, the PCA group (iii) consisted of samples that were characterized as invasive, keratinizing, and moderately differentiated, whereas group (ii) samples were mostly clinically characterized as nonkeratinizing. A total of 13/14 AA samples that tested p16-/RNA- possessed similar clinical attributes as samples classified as p16+/RNA-, suggesting that the p16 expression via immunohistochemistry test might be classifying a significant percent of HPV-negative AA samples as positive. Taken together, these results suggest that the p16 immunohistochemistry test for oropharyngeal cancer may be overly sensitive for classifying AA oropharynx cancer samples. Clinical decisions regarding treatment for AA based on this test have the potential to negatively impact outcomes. Citation Format: Batsirai M. Mabvakure, Shiting Li, Deborah de la Caridad Delgado Herrera, William Benjamin, Lucia Martinez Cruz, Siddhi Patil, Elham Mohebbi, Gregory T. Wolf, Anju Duttargi, Christina Lefante, Mei-Chin Hsieh, Nisha J. D'Silva, Maureen Sartor, Laura Rozek. Comparison of HPV virus-induced oropharyngeal cancer by self-reported race and the potential implications on treatment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB137.

Clinicopathologic and Molecular Characterization of High-Risk Human Papillomavirus-Positive Carcinomas of the Urinary Tract.

Human papillomavirus (HPV) is a well-known cause of squamous cell carcinomas of anogenital and oropharyngeal regions, where treatment strategies and prognosis depend on HPV status. The significance of HPV status in tumors arising along the urinary tract is not well established. To provide detailed clinical, morphologic, immunohistochemical, and molecular analysis of HPV+ urinary tract carcinomas (UTCs). We identified and retrospectively examined 12 HPV+ UTCs, confirmed by high-risk HPV in situ hybridization. The HPV+ UTCs originated from the urethra (9) and urinary bladder (3); 5 of 12 (42%) presented with nodal metastasis. On morphology, HPV+ UTCs were predominantly basaloid; well-differentiated squamous areas were focally seen. Available immunohistochemistry (IHC) showed strong staining for p16 (11 of 11), p63 (12 of 12), cytokeratin (CK) 903 (11 of 11), and CK5/6 (11 of 11); variable staining for GATA3 (8 of 12) and CK7 (4 of 11); and rare uroplakin II staining (1 of 12). Molecular analysis revealed the most frequently altered genes: KMT2C (42%), PIK3CA (42%), and KMT2D (25%). In contrast to published conventional urothelial and squamous cell carcinoma molecular data, TERTp mutation was rare (8%), and no TP53 or CDKN2A aberrations were identified. During available follow-up (11 of 12; median, 39 months), 6 patients required treatment for recurrence; ultimately, 1 died of disease, 2 were alive with disease, and 8 had no evidence of disease. Finally, we provide 11 HPV- squamous predominant UTCs for IHC and molecular comparisons; notably, a subset of HPV- UTCs was positive for p16 IHC (27%), making p16 IHC a less-specific surrogate marker for HPV status at this site. HPV+ UTCs show distinct clinical, morphologic, and molecular characteristics, suggesting important roles for HPV in UTC.

Relating pre-treatment non-Gaussian intravoxel incoherent motion diffusion-weighted imaging to human papillomavirus status and response in oropharyngeal carcinoma

Background and purposeDiffusion-weighted imaging (DWI) is a promising technique for response assessment in head-and-neck cancer. Recently, we optimized Non-Gaussian Intravoxel Incoherent Motion Imaging (NG-IVIM), an extension of the conventional apparent diffusion coefficient (ADC) model, for the head and neck. In the current study, we describe the first application in a group of patients with human papillomavirus (HPV)-positive and HPV-negative oropharyngeal squamous cell carcinoma. The aim of this study was to relate ADC and NG-IVIM DWI parameters to HPV status and clinical treatment response. Materials and methodsThirty-six patients (18 HPV-positive, 18 HPV-negative) were prospectively included. Presence of progressive disease was scored within one year. The mean pre-treatment ADC and NG-IVIM parameters in the gross tumor volume were compared between HPV-positive and HPV-negative patients. In HPV-negative patients, ADC and NG-IVIM parameters were compared between patients with and without progressive disease. ResultsADC, the NG-IVIM diffusion coefficient D, and perfusion fraction f were significantly higher, while pseudo-diffusion coefficient D* and kurtosis K were significantly lower in the HPV-negative compared to HPV-positive patients. In the HPV-negative group, a significantly lower D was found for patients with progressive disease compared to complete responders. No relation with ADC was observed. ConclusionThe results of our single-center study suggest that ADC is related to HPV status, but not an independent response predictor. The NG-IVIM parameter D, however, was independently associated to response in the HPV-negative group. Noteworthy in the opposite direction as previously thought based on ADC.

Radiomics Features in Predicting Human Papillomavirus Status in Oropharyngeal Squamous Cell Carcinoma: A Systematic Review, Quality Appraisal, and Meta-Analysis.

We sought to determine the diagnostic accuracy of radiomics features in predicting HPV status in oropharyngeal squamous cell carcinoma (SCC) compared to routine paraclinical measures used in clinical practice. Twenty-six articles were included in the systematic review, and thirteen were used for the meta-analysis. The overall sensitivity of the included studies was 0.78, the overall specificity was 0.76, and the overall area under the ROC curve was 0.84. The diagnostic odds ratio (DOR) equaled 12 (8, 17). Subgroup analysis showed no significant difference between radiomics features extracted from CT or MR images. Overall, the studies were of low quality in regard to radiomics quality score, although most had a low risk of bias based on the QUADAS-2 tool. Radiomics features showed good overall sensitivity and specificity in determining HPV status in OPSCC, though the low quality of the included studies poses problems for generalizability.

Utilizing the AJCC 8th Staging to Discriminate Survival Outcomes in HPV-associated Oropharyngeal Squamous Cell Carcinoma.

The American Joint Committee on Cancer (AJCC) staging 8th edition introduced major changes in the TNM staging of oropharyngeal squamous cell carcinoma (OPSCC) based on the human papillomavirus (HPV) status. This study aimed to observe how well the AJCC staging 8th edition precisely discriminates survival outcomes in patients with HPV-associated OPSCC using a large population database. Using the Surveillance, Epidemiology, and End Results database between 2010 and 2016, 7,448 patients with HPV-associated OPSCC were enrolled. Patients diagnosed with OPSCC and tested positive for HPV with information on the TNM staging according to the AJCC staging 7th edition were selected. Next, T-, N-, and clinical staging were reconstructed based on the AJCC staging 8th edition. Survival probabilities in both AJCC staging 7th and 8th editions were estimated and compared. Most patients (93.44%) were down-staged from the 7th to the 8th edition. The AJCC staging 8th edition showed more discriminatory power in predicting survival of patients with HPV-associated OPSCC than the AJCC staging 7th edition, regardless of the primary subsites. Additionally, clinical stage I patients with HPV-associated OPSCC according to the AJCC 8th edition showed better prognosis in case of high T staging than high N staging. Clinical staging according to the AJCC 8th edition compared to that of the 7th edition was an independent prognostic factor in patients with HPV-associated OPSCC. This study emphasizes the advantages of the new classification system for discriminating survival in HPV-associated OPSCC according to various factors.

Treatment Delay and HPV Status on OPSCC With Upfront Surgery: Analysis of National Cancer Database.

Evaluate the effect of treatment delay on survival in human papillomavirus (HPV)-positive and HPV-negative oropharyngeal squamous cell carcinoma (OPSCC) patients undergoing primary surgical resection. Retrospective cohort study using the 2010-2017 National Cancer Database. Multicenter database study. Patients >18 years old with OPSCC and known HPV status, treated surgically with or without postoperative radiation/chemotherapy were included. Two cohorts based on HPV status were grouped by time to treatment initiation (TD-TI, ≤30, 31-60, ≥61 days) and surgery to radiotherapy (TS-RT, ≤42, 43-66, ≥67 days). Univariate, Kaplan-Meier, and multivariate analyses assessed correlations between demographic and clinical factors with overall survival in treatment delay groups. Included were 1643 HPV-positive OPSCC patients and 391 HPV-negative OPSCC patients. No associations between survival and gender, age, race, insurance, or radiotherapy length were observed. Regardless of HPV status, larger tumor size (>2 cm) and lymphovascular invasion predicted worse survival. HPV negative patients with >4 lymph nodes involved had 2.5×greater mortality risk (P = .039). Robotic surgery was associated with improved survival only in HPV positive patients (hazard ratio [HR]: 0.41, P < .001). In HPV positive patients, higher TD-TI related to lower mean survival, although this was not significant on multivariate analysis. HPV negative patients with >42 days of TS-RT had decreased survival (43-66 days, HR 1.63, P = .049; ≥67 days, HR 2.10, P = .032). Longer TS-RT was associated with lower overall survival in HPV negative patients. Treatment delay was not associated with survival in HPV positive OPSCC according to multivariate analysis. These findings enhance knowledge about treatment delay effects in OPSCC, aiding providers in decisions and patient communication.

The importance of combined HPV and CINtec® PLUS genotyping testing for p16 in women with cervical squamous cell carcinoma.

Immunohistochemistry (IHC) for p16INK4A (p16) is a reliable surrogate test for the presence of a high-risk, potentially transformative human papillomavirus (HPV) infection in precursor and malignant lesions of the cervix. The purpose of this study was to evaluate changes in cervical cells caused by persistent HPV infection, by IHC (p16 protein) by comparison with HPV genotyping. The study included female patients aged between 26 and 57 years who presented to a public hospital, with complaints related to the genital area, namely vaginal bleeding and dyspareunia. After selecting the patients, samples were subjected to cytological testing and IHC for p16 and for the determination of HPV types. The relationship between HPV status and p16 status was statistically significant (p=0.0001), of the 41 patients, 53.7% were HPV positive, respectively 56.1% were p16 positive, the agreement relationship between the two indicators was very high (weighted kappa: 0.951). The clinical performance of CINtec® PLUS triage for p16 shows a high positive predictive value (PPV) and a high negative predictive value (NPV) of 95.7% and 100%, respectively, as regards HPV. The p16 marker (CINtec® PLUS) can be used as a prognostic biomarker and provides clinical usefulness through increased sensitivity (Se) and specificity (Sp) in the triage of women at risk of developing precancerous lesions, compared to cytology that is based on morphology, but has a rather low Se and high Sp, while HPV testing is very sensitive but slightly more specific.

Abstract 2501: High mutation burden and ultraviolet light signature in conjunctival squamous cell carcinoma

Abstract Purpose: Conjunctival squamous cell carcinoma (conjSCC) is endemic in sub-Saharan Africa and is a major cause of ocular morbidity and mortality. This study aims to evaluate Tumor Mutation Burden (TMB), genomic signature, and histologic characteristics of conjunctival squamous cell carcinoma (conjSCC) in two cohorts of patients; one from the USA and one from Ethiopia. Methods: Whole exome sequencing data, PD-L1 expression, histologic features, and human papillomavirus (HPV) status were analyzed in conj SCC tumors of 25 patients treated at an Ethiopian (ETH) cancer center and 29 patients treated at a US cancer center. Genomic alterations in conjSCC were compared with SCCs from other anatomic sites using data from The Cancer Genome Atlas (TCGA). Results: TMB was found to be relatively high in conjunctival SCC (3.01/Mb, log10), was comparable to TMB values associated with cutaneous SCC, and was higher than most other SCC types available in the TCGA database. ETH samples had higher tumor mutational burden compared to the US cohort (3.34 vs 2.73). Solar elastosis was seen in a higher percentage of tumors from Ethiopia compared with the US cohort (78% vs. 10%). HPV status was similar between the cohorts (ETH: 21%, MDA: 28%). PD-L1 positivity defined as &amp;gt; 1% of tumor cells was seen in 33 of 53 tumors (62%). Mutations associated with ultraviolet light (UV) signature were the most frequently encountered signatures in both cohorts (SBS7b: 74% and SBS7a: 72%). UV signatures were more prevalent in the ETH cohort compared with the US cohort. Conclusions: Conjunctival SCC is associated with a high TMB providing the opportunity for immune checkpoint inhibitor therapy for patients with locally advanced and/or metastatic conjunctival SCC. Clinical trials are needed to assess the efficacy of immune checkpoint inhibitors in patients with conjunctival SCC, particularly in sub-Saharan Africa where this cancer is more frequently seen, and is associated with significant morbidity and mortality. Citation Format: Bita Esmaeli, Frederico O. Gleber-Netto, Oded Sagiv, Curtis R. Pickering, Neil Gross, Jing Ning, Melisachew M. Yeshi, Jonas Mitku, Michael T. Tetzlaff, Priyadharsini Nagarajan. High mutation burden and ultraviolet light signature in conjunctival squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2501.

Abstract 4403: The crosstalk between the tumor microbiome and epigenomic changes in penile squamous cell carcinoma

Abstract Penile squamous cell carcinoma (PSCC) is approximately three times more prevalent in Puerto Rico when compared to other ethnic groups in the United States. Moreover, the mortality rate for PSCC among Puerto Rican men is significantly higher when compared to the US population. Infection with human papillomavirus (HPV) has been identified as a risk factor for an average of 48% of PSCC cases in Puerto Rico. The molecular etiology of HPV+ and HPV- PSCC remains poorly understood. To date, the role of the microbiota in the pathogenesis of PSCC is unknown, as there are no studies that characterize the microbiome in association with penile cancer. Furthermore, the role of DNA methylation, which is an epigenetic alteration that can be found in cancer cells that can lead to changes in gene expression and cellular function, is also unknown. Therefore, there is an urgent need to address this knowledge gap. This study aims to analyze the DNA methylation patterns and bacterial communities in HPV-positive and HPV-negative PSCC tissues from Puerto Rican men. DNA was extracted from twenty-one fresh PSCC tissue samples from Puerto Rican patients. HPV status and genotyping were determined using the DNA ELISA kit HPV SPF10 protocol. DNA methylation assay was performed using the Infinium Methylation EPIC v2.0 BeadChip Kit to identify and compare average DNA methylation levels (%) in 7 HPV-positive and 14 HPV-negative penile cancer samples. Microbiota analyses were performed using 16S rRNA genes with the Illumina MiSeq platform. Demultiplexed data was deposited in QIITA for quality control and bioinformatic analyses and downstream analyses were done in R and Qiime2. Our results demonstrated that 45 loci methylation patterns were significantly different between HPV-positive and HPV-negative samples, with an adjusted p-value &amp;lt;0.1 (Benjamini-Hochberg method). Three loci methylation patterns were significantly affected by species diversity and richness. The abundance of Firmicutes, Proteobacteria, Bacteroidetes, Fusobacteria, and Actinobacteria was individually associated with number of affected loci (27, 19, 12, 6, and 3, respectively). We found that HPV status is associated with methylation patterns and microbiome in PSCC in Puerto Rican men. Both changes may be involved in triggering inflammatory responses and oncogenesis. Although many challenges must be overcome to dissect the specific interactions of coinfecting bacteria and methylation patterns during the penile cancer infectious process, our findings demonstrate that microbes may be involved in these cellular processes. Citation Format: Jorge Viera-Vera, Yarelis M. Roque-Reyes, Daniel Santiago-Negron, María Sánchez-Vázquez, Yakshi Ortiz Maldonado, Gabriel Borges-Vélez, Jasmine Figueroa-Díaz, Carlos A. Rivera-López, Niki M. Zacharias Millward, María J. Marcos-Martínez, Filipa Godoy-Vitorino, Antonio Puras-Baez, Julie Dutil, Josué Pérez-Santiago, Magaly Martínez-Ferrer. The crosstalk between the tumor microbiome and epigenomic changes in penile squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4403.

Abstract 6300: Associations of biological aging with perceived stress in patients with head and neck cancer: A longitudinal study

Abstract Introduction: Biological aging leads to an increased risk of morbidity and mortality in patients with head and neck cancer (HNC) undergoing intensity-modulated radiotherapy (IMRT). This progression can be exacerbated by the burden of stress arising from HNC and IMRT. While stress has been linked to inflammation, the associations of biological aging measures including inflammatory markers and epigenetic age acceleration (EAA) with stress over cancer treatment remain underexplored. The purpose of this study was to examine the associations between biological aging and stress in patients with HNC receiving IMRT across four time points (i.e., pre- (T1), end of (T2), 3 months (T3), and 12 months (T4) post-IMRT). Methods: We conducted a prospective longitudinal study. Stress was assessed using the Perceived Stress Scale (PSS) across four time points along with the assessment of biological aging markers: peripheral inflammatory markers (i.e., interleukin [IL-6], IL-1β, IL-10, IL-1ra, C-reactive protein [CRP], soluble tumor necrosis factor receptor 2 [sTNFR2], and TNF-α) and EAA. EAA was calculated using DNAmPhenoAge, adjusted for chronological age. Liner mixed effects models were used to examine the associations between biological aging measures and stress across time. Results: A total of 176 patients with HNC were enrolled in the study with a mean age of 59.9±10.0 years, 72.7% male, and 77.3% non-Hispanic White. Stress scores increased from T1 to T2 (p&amp;lt;0.001), decreased significantly at T3 (p&amp;lt;0.001) which were lower than the baseline scores (T1), and then gradually decreased at T4 (p=0.029). Significant associations of CRP, IL- 1ra, and TNFR-2 with stress were identified across time (all p values &amp;lt;0.05), controlling for covariates (i.e., age, human papillomavirus status, chemotherapy, and surgery): elevated inflammatory markers were associated with higher levels of stress. In a separate model, higher stress scores were also associated with increased EAA controlling for the same covariates (p=0.015): an increment of 10 units in stress scores was associated with an age acceleration of 7.2 months across time. Conclusions: Findings from our study suggest that biological aging measures (inflammatory markers and EAA) are associated with stress across time in patients with HNC receiving IMRT. Clinicians and researchers may use the findings to develop personalized psychosocial interventions that reduce stress so as to deaccelerate biological aging for patients with HNC during their treatment and survivorship. Citation Format: Yufen Lin, Telisa Spikes, Deborah Bruner, Sudeshna Paul, Andrew Miller, Karen Conneely, Nabil Saba, Canhua Xiao. Associations of biological aging with perceived stress in patients with head and neck cancer: A longitudinal study [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6300.

Abstract 2297: Longitudinal monitoring of HPV circulating tumor DNA in cervical cancer

Abstract Introduction Detecting circulating tumor DNA (ctDNA) in the blood using minimally invasive techniques known as a "liquid biopsy", is gaining tremendous popularity in oncology research. Integration of human papillomavirus (HPV) DNA and the overexpression of the E6 and E7 oncogenes are crucial steps in the development of cervical cancer. We investigated the potential utility of HPV ctDNA as a biomarker for monitoring treatment outcomes in patients with cervical cancer. Material and Methods We use droplet digital PCR (ddPCR) to measure the amount of HPV16/18 ctDNA in plasma of cervical cancer patients. The HPV standard GP5+/6+ primers were used to determine HPV positivity in our patients. Specific primers were designed to amplify specifically the E6 oncogene of HPV16 and HPV18. Plasmid constructs were made using the PCR-purified products of HeLa and SiHa cell lines which would act as a positive control for the assay. HPV16 and 18 plasmids were serially diluted and then tested in qPCR as a template using different primer sets. Standard curves were made from the recombinant plasmids. This will serve as a positive control and will also provide a correlation between the sensitivity and specificity of qPCR vs ddPCR. Results and Discussions We hypothesize that our dPCR test will be able to detect minute quantities of HPV ctDNA in plasma of cervical cancer patients as ctDNA levels are expected to be decrease as treatment progresses. The assay sensitivity and specificity will be obtained by comparing results to QPCR. The study samples involve a tumor biopsy and a blood sample at the baseline prior to the therapy and serial blood sampling in different stages of treatment and follow-ups. These samples will also serve for treatment monitoring of these patients through ddPCR. Tumor samples are used to confirm the HPV status of patients using HPV-specific GP5+/GP6+ qPCR. We have recruited 75 cervical cancer patients with majority of them being squamous cell carcinoma and stage IIB disease. The qPCR results from tumor biopsy revealed HPV 16 was most prevalent (32/43), followed by HPV18 (6/43) and HPV 33(2/43). Conclusion we will now use Qiagen digital PCR system to understand HPV ctDNA levels in our patient cohort. we believe that our comprehensive sample collection after every treatment milestone and thorough follow-up for upto 2 years after diagnosis will allow us to study the utility of this sensitive test in monitoring disease and relapse. Citation Format: Preetiparna Parida, Krishna Sharan, Shirley Lewis, Rama Rao Damerla. Longitudinal monitoring of HPV circulating tumor DNA in cervical cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2297.

Impact of 18F-FDG PET Intensity Normalization on Radiomic Features of Oropharyngeal Squamous Cell Carcinomas and Machine Learning-Generated Biomarkers.

We aimed to investigate the effects of 18F-FDG PET voxel intensity normalization on radiomic features of oropharyngeal squamous cell carcinoma (OPSCC) and machine learning-generated radiomic biomarkers. Methods: We extracted 1,037 18F-FDG PET radiomic features quantifying the shape, intensity, and texture of 430 OPSCC primary tumors. The reproducibility of individual features across 3 intensity-normalized images (body-weight SUV, reference tissue activity ratio to lentiform nucleus of brain and cerebellum) and the raw PET data was assessed using an intraclass correlation coefficient (ICC). We investigated the effects of intensity normalization on the features' utility in predicting the human papillomavirus (HPV) status of OPSCCs in univariate logistic regression, receiver-operating-characteristic analysis, and extreme-gradient-boosting (XGBoost) machine-learning classifiers. Results: Of 1,037 features, a high (ICC ≥ 0.90), medium (0.90 > ICC ≥ 0.75), and low (ICC < 0.75) degree of reproducibility across normalization methods was attained in 356 (34.3%), 608 (58.6%), and 73 (7%) features, respectively. In univariate analysis, features from the PET normalized to the lentiform nucleus had the strongest association with HPV status, with 865 of 1,037 (83.4%) significant features after multiple testing corrections and a median area under the receiver-operating-characteristic curve (AUC) of 0.65 (interquartile range, 0.62-0.68). Similar tendencies were observed in XGBoost models, with the lentiform nucleus-normalized model achieving the numerically highest average AUC of 0.72 (SD, 0.07) in the cross validation within the training cohort. The model generalized well to the validation cohorts, attaining an AUC of 0.73 (95% CI, 0.60-0.85) in independent validation and 0.76 (95% CI, 0.58-0.95) in external validation. The AUCs of the XGBoost models were not significantly different. Conclusion: Only one third of the features demonstrated a high degree of reproducibility across intensity-normalization techniques, making uniform normalization a prerequisite for interindividual comparability of radiomic markers. The choice of normalization technique may affect the radiomic features' predictive value with respect to HPV. Our results show trends that normalization to the lentiform nucleus may improve model performance, although more evidence is needed to draw a firm conclusion.