Pathogens

Gut–Vaginal Microbiome Crosstalk in Ovarian Cancer: Implications for Early Diagnosis

Ovarian cancer remains a formidable global health burden, characterized by frequent late-stage diagnosis and elevated mortality rates attributable to its elusive pathogenesis and the critical lack of reliable early-detection biomarkers. Emerging investigations into the gut–vaginal microbiome axis have unveiled novel pathogenic mechanisms and potential diagnostic targets in ovarian carcinogenesis. This comprehensive review systematically examines the compositional alterations in and functional interplay between vaginal and intestinal microbial communities in ovarian cancer patients. We elucidate three principal mechanistic pathways through which microbial dysbiosis may drive oncogenesis: (1) estrogen-mediated metabolic reprogramming via β-glucuronidase activity; (2) chronic activation of pro-inflammatory cascades (particularly NF-κB and STAT3 signaling); (3) epigenetic silencing of tumor suppressor genes through DNA methyltransferase modulation. We propose an integrative diagnostic framework synthesizing multi-omics data—incorporating microbial profiles, metabolic signatures, pathway-specific molecular alterations, established clinical biomarkers, and imaging findings—within a multifactorial etiological paradigm. This innovative approach aims to enhance early-detection accuracy through machine learning-enabled multidimensional pattern recognition. By bridging microbial ecology with tumor biology, this review provides novel perspectives for understanding ovarian cancer etiology and advancing precision oncology strategies through microbiome-targeted diagnostic innovations.

Trends in Human Papillomavirus-Related Health Burden in Greece from 1996 to 2021 with a Focus on Cervical and Lip, Oral Cavity, and Pharyngeal Cancer

This study aimed to evaluate the burden of HPV-related hospitalization and mortality in Greece, with a focus on invasive cervical cancer and lip, oral cavity, and pharyngeal (LOCP) cancers. A retrospective query using data from the Greek Statistical Office and Eurostat was executed. The query included hospital admission and standardized mortality rates (SDRs) on cervical dysplasia and cervical, vulvar, and vaginal; anal; penile; and LOCP cancers. The hospitalization rate for invasive cervical cancer decreased over time, exhibiting a sharp decrease after 2010, while the hospitalization rate for LOCP cancer decreased after 2011, preceded by a sustained increase from 1996. The hospitalization rate of HPV-attributable diseases in total showed a declining tendency between 2013 and 2017. SDR due to cervical cancer showed a slightly decreasing trend in Greece and the European Union, while SDR due to LOCP cancer showed a slightly increasing trend in Greece, but a decrease in the European Union. The decline in hospitalization rates for HPV-related disease in Greece, especially for cervical cancer and dysplasia, and also the declining SDR for invasive cervical cancer in Greece and the EU, are indications of the positive public health impact of screening programs and the implementation of HPV vaccination.

Molecular Features of HPV-Independent Cervical Cancers

HPV-independent cervical cancers represent a small proportion of these types of cancers, predominantly glandular lesions. It should be noted that some cases may depend on diagnostic problems that lead to false negative cases. However, the most recent classifications distinguish cervical tumors into HPV-associated and HPV-independent cancers. HPV-negative cervical carcinomas (5–11% of all cases) mainly include rare adenocarcinomas (gastric, mesonephric, clear, serous, and endometrioid) and present distinct clinical and molecular features. These tumors usually affect older women and are diagnosed at more advanced stages than HPV-positive tumors, with an overall worse prognosis. This concerning and notably worse prognosis highlights the need for further research and understanding. Unlike HPV-positive carcinomas (which depend on the viral oncogenes E6/E7), HPV-independent tumors accumulate genomic mutations that activate oncogenes and inactivate suppressor genes. Therefore, a comprehensive overview of these aspects can be the key to a better understanding and developing personalized treatments. In the present review, the main mutated genes, the signaling pathways involved, the differences from HPV-positive tumors, the distinctive immunohistochemical markers, and the diagnostic and therapeutic implications are explored in depth.

Genotype-Specific HPV E6/E7 mRNA Triage Improves Risk Stratification and Reduces Referrals in DNA-Positive ASC-US/LSIL: A Real-World Cohort from Nordland, Norway

HPV DNA–positive women with ASC-US/LSIL cytology represent a heterogeneous risk group in cervical screening and require efficient triage. We evaluated a genotype-specific 7-type HPV E6/E7 mRNA assay (PreTect HPV-Proofer 7; types 16/18/31/33/45/52/58) in a real-world quality-assurance cohort at Nordland Hospital (Bodø, Norway). Among HPV-positive women with ASC-US/LSIL reflex cytology, 225 had sufficient residual liquid-based cytology material and a valid mRNA result; 175 had complete follow-up (2022–2025) and were included. Overall, 44.6% (78/175) were mRNA-positive (ASC-US 45.2%; LSIL 43.3%). For CIN2+, sensitivity was 63.4%, specificity 61.2%, PPV 33.3%, and NPV 84.5%; CIN2+ risk was 33.3% in mRNA-positive versus 15.5% in mRNA-negative women (RR 2.16, 95% CI 1.23–3.78). For CIN3+, risk was 14.1% versus 6.2%. Genotype-specific PPVs were highest for HPV33, HPV18, HPV16, and HPV31. In a referral simulation, mRNA-guided triage reduced baseline colposcopy referrals by 55% and decreased colposcopies per detected CIN2+ by ~30%, while 15 CIN2+ and 6 CIN3+ occurred in the mRNA-negative group and would be expected to be detected at 12-month follow-up among women with persistent HPV positivity. Genotype-aware HPV E6/E7 mRNA triage improves risk stratification and may increase screening efficiency.

Patients with Newly Diagnosed Cervical Cancer Should Be Screened for Anal Human Papillomavirus (HPV) and Anal Dysplasia: Results of Cost and Quality Analysis

Background: HPV infections with high-risk subtypes are a risk factor for developing cervical and anal cancer. Despite HPV vaccination, the incidence of cervical and anal cancer is increasing. There has been substantial research regarding the benefits of screening men who have sex with men (MSM) and those diagnosed with HIV for anal HPV and dysplasia, but little data exists for women diagnosed with cervical cancer. Methods: We constructed a Markov model in Python 3.13 to simulate the outcomes and financial impact of screening women newly diagnosed with cervical cancer for anal HPV and dysplasia. Two matrices were used to represent the screened group and the unscreened group. In the screening group, all women received initial anal HPV screening and high-resolution anoscopy with biopsy. If biopsy results confirmed HSIL, each would receive treatment with electrocautery. The screening group would also undergo annual surveillance and follow-up treatment as necessary. In the unscreened group, women did not receive screening or treatment, and the disease process was allowed to progress naturally. Results: The initial cohort consisted of 5555 women diagnosed with cervical cancer and concurrent anal HPV. The incidence of anal cancer in the screening group was 271 vs. 375 in the unscreened group after three years, 642 vs. 1236 after ten years, and 863 vs. 2039 after twenty years. Moreover, anal cancer deaths were 1236 in the screening group vs. 9041 in the unscreened group after 10 years and 31,118 vs. 51,553 after twenty years. The screened group saved 330.1 million dollars after ten years and 1.33 billion dollars after twenty years when compared to the unscreened group. Over the life of the study, the screened group would also accrue 102,000 discounted QALYs when compared to the unscreened group. Conclusions: Our model strongly suggests that screening women diagnosed with cervical cancer for anal HPV and treating anal dysplasia leads to less anal cancer, less deaths from anal cancer, less economic impact on the healthcare system, and more QALYs for patients.

Unlocking the Interactions Between the Whole-Body Microbiome and HPV Infection: A Literature Review

The human microbiome plays a vital role in maintaining human homeostasis, acting as a key regulator of host immunity and defense mechanisms. However, dysbiotic microbial communities may cause disruption of the symbiotic relationship between the host and the local microbiota, leading to the pathogenesis of various diseases, including viral infections and cancers. One of the most common infectious agents causing cancer is the human papilloma virus (HPV), which accounts for more than 90% of cervical cancers. In most cases, the host immune system is activated and clears HPV, whereas in some cases, the infection persists and can lead to precancerous lesions. Over the last two decades, the advent of next-generation sequencing (NGS) technology and bioinformatics has allowed a thorough and in-depth analysis of the microbial composition in various anatomical niches, allowing researchers to unveil the interactions and the underlying mechanisms through which the human microbiota could affect HPV infection establishment, persistence, and progression. Accordingly, the present narrative review aims to shed light on our understanding of the role of the human microbiome in the context of HPV infection and its progression, mainly to cervical cancer. Furthermore, we explore the mechanisms by which the composition and balance of microbial communities exert potential pathogenic or protective effects, leading to either HPV persistence and disease outcomes or clearance. Special interest is given to how the microbiome can modulate host immunity to HPV infection. Lastly, we summarize the latest findings on the therapeutic efficacy of probiotics and prebiotics in preventing and/or treating HPV infections and the potential of vaginal microbiota transplantation while highlighting the significance of personalized medicine approaches emerging from NGS-based microbiome profiling and artificial intelligence (AI) for the optimal management of HPV-related diseases.

Post-Conization HPV Vaccination and Its Impact on Viral Status: A Retrospective Cohort Study in Troms and Finnmark, 2022

Human papillomavirus (HPV) is associated with cellular changes in the cervix leading to cancer, which highlights the importance of vaccination in preventing HPV infections and subsequent cellular changes. Women undergoing the loop electrosurgical excision procedure (LEEP), a treatment for high-grade cervical intraepithelial neoplasia (CIN2+), remain at risk of recurrence. This study assessed the effect of post-conization HPV vaccination on the viral status of women at six months post-conization, aiming to evaluate the vaccine’s effectiveness in preventing recurrence of CIN2+. A retrospective cohort study was conducted among women in Troms and Finnmark who underwent conization in 2022. Using the SymPathy database and the national vaccination register (SYSVAK), we analyzed the vaccination statuses and HPV test results of women born before 1991, who had not received the HPV vaccine prior to conization. Out of 419 women undergoing conization, 243 met the inclusion criteria. A significant association was found between post-conization HPV vaccination and a negative HPV test at six months of follow-up (ARR = 12.1%, p = 0.039). Post-conization HPV vaccination significantly reduced the risk of a positive HPV test at the first follow-up, suggesting its potential in preventing the recurrence of high-grade cellular changes. However, the retrospective design and the insufficient control of confounding variables in this study underscore the need for further studies to confirm these findings.

Genotype-Specific HPV mRNA Triage Improves CIN2+ Detection Efficiency Compared to Cytology: A Population-Based Study of HPV DNA-Positive Women

Background: Effective triage of women testing positive for high-risk HPV DNA is essential to reduce unnecessary colposcopies while preserving cancer prevention. Cytology, the current standard, has limited specificity and reproducibility. The genotype-specific 7-type HPV E6/E7 mRNA test (PreTect HPV-Proofer’7), targeting HPV types 16, 18, 31, 33, 45, 52, and 58, detects transcriptionally active infections and may enhance risk stratification. Methods: Between 2019 and 2023, 34,721 women aged 25–69 underwent primary HPV DNA screening with the Cobas 4800 assay at the University Hospital of North Norway, within the national screening program. Of these, 1896 HPV DNA-positive women were triaged with liquid-based cytology with atypical squamous cells of undetermined significance or worse (≥ASC-US) and the 7-type HPV mRNA test. Histological outcomes were followed through October 2024. Diagnostic performance for CIN2+ was evaluated overall and by genotype. Results: CIN2+ prevalence was 13.3%. The mRNA test reduced test positivity from 50.3% to 33.4% while maintaining comparable sensitivity (70.6% vs. 72.2%) and improving specificity (72.3% vs. 53.0%) and PPV (28.1% vs. 19.1%). Genotype-specific PPVs were highest for HPV16 mRNA (47.7%), followed by HPV33 (39.2%) and HPV31 (32.2%), all exceeding corresponding DNA-based estimates. Conclusion: Genotype-specific HPV mRNA triage offers superior risk discrimination compared to cytology, supporting more targeted, efficient, and accessible cervical cancer screening.

Global Burden and Incidence Trends in Cancers Associated with Human Papillomavirus Infection: A Population-Based Systematic Study

Background: Human papillomavirus (HPV) is responsible for a substantial fraction of anogenital and head and neck cancers (HNC). HPV-related cancers cause a heavy burden globally, with disparities across different cancers. We aimed to present an up-to-date global view of the patterns and incidence trends among HPV-related cancers. Methods: We collected data on HPV-related cancers from the GLOBOCAN 2022 database and the Cancer Incidence in Five Continents plus Compendium. Age-standardized incidence and mortality rate (ASIR and ASMR) were calculated to estimate the cancer burden. Spearman’s correlation tests were used to evaluate the associations with the Human Development Index (HDI). Joinpoint regression was conducted to evaluate the incidence trends in ASIR. Results: In 2022, 1,505,394 HPV-related cancer cases and 755,303 deaths were newly estimated worldwide, corresponding to an overall ASIR and ASMR of 20.9 and 10.2 per 100,000 people, respectively. Africa had the highest ASIR and ASMR compared with Asia, accounting for the most new cases and deaths. The primary cause was cervical cancer (ASIR 14.1 per 100,000 people); however, HNC exhibited the largest number of cases (685,204 cases). The total rates of HPV-related cancers were 1.3 times higher for ASIR and nearly three times higher for ASMR in low-HDI countries than in very high-HDI countries. A decreasing trend was observed for the ASIR of cervical cancer in most studied countries, compared to the increasing trends in HNC in females and anal cancer in both sexes. Conclusions: The global burden and trends of HPV-related cancers vary significantly among different cancer types according to region and sex. Particularly, cervical, HNC, and anal cancers should attract global attention. However, specific cancer types contributing to the heaviest burden should be identified at the country level to adjust resource allocation and improve access to quality health services.

FAM19A4 and hsa-miR124-2 Double Methylation as Screening for ASC-H- and CIN1 HPV-Positive Women

The DNA methylation levels of host cell genes increase with the severity of the cervical intraepithelial neoplasia (CIN) grade and are very high in cervical cancer. Our study aims to evaluate FAM19A4 and hsa-miR124-2 methylation in Atypical Squamous cells with high-grade squamous intraepithelial lesions (ASC-H) and in CIN1, defined as low-grade squamous intraepithelial lesions (LSILs) by the Bethesda classification, as possible early warning biomarkers for managing women with high-risk HPV infections (hrHPV). FAM19A4 and hsa-miR124-2 methylation tests were conducted on fifty-six cervical screening samples from a subset of women aged 30–64 years old. Specimens were collected into ThinPrep PreservCyt Solution. Their HrHPV genotype and cytology diagnosis were known. A Qiasure (Qiagen) was used for FAM19A4 and hsa-miR124-2 methylation testing on bisulfite-converted DNA, according to the manufacturer’s specifications. The reported results were hypermethylation-positive or -negative. We found that FAM194A4 and hsa-miR124-2 methylation was detected in 75% of ASC-H cases with a persistent infection of hrHPV. A total of 60% of CIN1 lesions were found to be positive for methylation, and 83.3% were when the cytology was CIN2/3. In addition, as a novelty of this pilot study, we found that combined FAM19A4 and hsa-miR124-2 methylation positivity rates (both methylated) were associated with the HPV genotypes 16, 18, and 59 and covered 22 and 25% of ASC-H and CIN1 cases, respectively. The methylation of these two genes, in combination with HPV genotyping, can be used as an early warning biomarker in the management and follow-up of women with ASC-H and CIN1 to avoid their progression to cervical cancer.

Unveiling the Dual Threat: How Microbial Infections and Healthcare Deficiencies Fuel Cervical and Prostate Cancer Deaths in Africa

Cervical and prostate cancer account for 7.1 and 7.3 deaths per 100,000 people globally in 2022. These rates increased significantly to 17.6 and 17.3 in Africa, respectively, making them the second and third leading cause of cancer deaths in Africa, only surpassed by breast cancer. The human papillomavirus is the prime risk factor for cervical cancer infection. On the other hand, prostate cancer risks include ageing, genetics, race, geography, and family history. However, these factors alone cannot account for the high mortality rate in Africa, which is more than twice the global mortality rate for the two cancers. We searched PubMed, Embase, Scopus, and Web of Science to select relevant articles using keywords related to microorganisms involved in cervical and prostate cancer and the impact of poor healthcare systems on the mortality rates of these two cancers in Africa by carrying out a detailed synopsis of the studies on microbial agents involved and the contributory factors to the deteriorating healthcare system in Africa. It became apparent that the developed countries come first in terms of the prevalence of cervical and prostate cancer. However, more people per capita in Africa die from these cancers as compared to other continents. Also, microbial infections (bacterial or viral), especially sexually transmitted infections, cause inflammation, which triggers the pathogenesis and progression of these cancers among the African population; this has been linked to the region’s deficient health infrastructure, making it difficult for people with microbial infections to access healthcare and hence making infection control and prevention challenging. Taken together, untreated microbial infections, primarily sexually transmitted infections due to the deficient healthcare systems in Africa, are responsible for the high mortality rate of cervical and prostate cancer.

Human Papillomavirus (HPV) Infection and Risk Behavior in Vaccinated and Non-Vaccinated Paraguayan Young Women

Cervical cancer is a global health concern and ranks fourth among the most prevalent cancers in women worldwide. Human papillomavirus (HPV) infection is a known precursor of cervical cancer and preventive measures include prophylactic vaccines. This study focused on sexually active Paraguayan women aged 18–25 years, exploring the intersection of HPV vaccination and sexual behavior. Among 254 participants, 40.9% received the Gardasil-4 vaccine, with no significant differences in sexual behavior between the vaccinated and unvaccinated sexually active groups. However, a notable decrease in the prevalence of HPV among the vaccinated women highlights the efficacy of this vaccine in reducing infections. The prevalence of any HPV type was 37.5% in vaccinated participants compared to 56.7% in unvaccinated participants (p = 0.0026). High-risk HPV types showed a significant difference, with a prevalence of 26.0% in vaccinated women compared with 52.7% in unvaccinated women (p < 0.001). Although a potential decline in genital warts was observed among the vaccinated individuals, statistical significance (p = 0.0564) was not reached. Despite the challenges in achieving high vaccination coverage, the observed reduction in HPV prevalence underscores the importance of ongoing monitoring, healthcare professional recommendations, and comprehensive risk management. These findings contribute to dispelling concerns about HPV vaccination influencing sexual behavior, advocating further large-scale research to explore the impact of vaccines on various HPV types and potential cross-protection.

Evaluation of the PreTect HPV-Proofer E6/E7 mRNA Assay for the Detection of Precancerous Cervical Lesions in the Greek Female Population

Cervical cancer remains a significant public health concern, ranking as the 10th most common cancer among women in Greece. Current screening programs primarily rely on cytology and HPV DNA testing; however, their positive predictive value (PPV) for detecting high-grade cervical intraepithelial neoplasia (CIN2+) remains limited. This study aimed to compare the clinical performance of the HPV mRNA test with that of the HPV DNA test, focusing on their PPV for detecting CIN1+ lesions in a cohort of Greek women. A total of 114 women undergoing routine cervical cancer screening were tested using an HPV DNA assay (detecting 41 HPV types), Pap smear, and were referred for colposcopy and biopsy when indicated. Among them, 29 women aged 18 to 65 years (mean age: 35.1 ± 10.8 years) who tested positive for one or more of the five high-risk HPV types (16, 18, 31, 33, 45) were further assessed using the PreTect HPV-Proofer® mRNA assay. Of these 29 women, 11 (37.9%) had negative biopsy findings, 16 (55.2%) were diagnosed with CIN1, and 2 (6.9%) with CIN2, corresponding to a positive predictive value (PPV) of 55.2% for CIN1 and 6.9% for CIN2 with the HPV DNA test. Among the 17 women who tested positive for HPV mRNA, 13 were diagnosed with CIN1 and 2 with CIN2. Among the 12 women who tested negative for HPV mRNA, 3 had CIN1 and 9 had negative biopsy results. Based on these findings, the PPV of the HPV mRNA test for CIN1 was 76.5%, the negative predictive value (NPV) was 75.0%, and the clinical sensitivity for CIN1 was 81.3%. For CIN2, the PPV was 11.8%, while the clinical sensitivity and NPV were 100%. These findings highlight the potential of HPV mRNA testing to improve specificity in cervical cancer screening by more accurately identifying clinically significant lesions and reducing unnecessary colposcopies.

Oncolytic Viruses in Ovarian Cancer: Where Do We Stand? A Narrative Review

Ovarian cancer (OC) remains the most lethal gynecologic malignancy with limited effective treatment options. Oncolytic viruses (OVs) have emerged as a promising therapeutic approach for cancer treatment, capable of selectively infecting and lysing cancer cells while stimulating anti-tumor immune responses. Preclinical studies have demonstrated significant tumor regression and prolonged survival in OC models using various OVs, such as herpes simplex. Early-phase clinical trials have shown a favorable safety profile, though the impact on patient survival has been modest. Current research focuses on combining OVs with other treatments like immune checkpoint inhibitors to enhance their efficacy. We provide a comprehensive overview of the current understanding and future directions for utilizing OVs in the management of OC.