Background: Cervical cancer is caused by Human Papilloma viruses (HPV) and is preceded by precursor stages: Cervical Intraepithelial Neoplasia (CIN). CIN is mostly found in women in their reproductive age and treated with a Loop Electrosurgical Excision Procedure (LEEP). The recurrence or residual disease rate after treatment is up to 17%. These women have a lifelong increased risk of recurrent CIN, cervical cancer and other HPV related malignancies. Furthermore, LEEP treatments are associated with complications such as premature birth. Limited data show that prophylactic HPV vaccination at the time of LEEP reduces recurrence rates, therefore leading to a reduction in repeated surgical interventions and side effect like preterm birth. The primary study objective is to evaluate the efficacy of the nonavalent HPV vaccination in women with a CIN II-III (high-grade squamous intraepithelial lesion (HSIL) lesion who will undergo a LEEP in preventing recurrent CIN II-III after 24 months. Methods: This study is a randomised, double blinded, placebo controlled trial in 750 patients without prior HPV vaccination or prior treatment for CIN and with histologically proven CIN II-III (independent of their hrHPV status) for whom a LEEP is planned. Included patients will be randomised to receive either three injections with nonavalent (9 HPV types) HPV vaccine or placebo injections (NaCL 0.9%) as a comparator. Treatment and follow-up will be according the current Dutch guidelines. Primary outcome is recurrence of a CIN II or CIN III lesion at 24 months. A normal PAP smear with negative hrHPV test serves as surrogate for absence of CIN. At the start and throughout the study HPV typing, quality of life and cost effectiveness will be tested. Discussion: Although prophylactic HPV vaccines are highly effective, little is known about the effectivity of HPV vaccines on women with CIN. Multiple LEEP treatments are associated with complications. We would like to evaluate the efficacy of HPV vaccination in addition to LEEP treatment to prevent residual or recurrent cervical dysplasia and decrease risks of repeated surgical treatment. Trial registration: Medical Ethical Committee approval number: NL66775.078.18. Affiliation: Erasmus Medical Centre. Dutch trial register: NL 7938. Date of registration 2019-08-05.

This study aims to refine our understanding of the inherent heterogeneity in cervical cancer by exploring differential gene expression profiles, immune cell infiltration dynamics, and implicated signaling pathways in the two predominant histological types of cervix carcinoma, Squamous Cell Carcinoma (SCC) and Adenocarcinoma (ADC). Targeted gene expression data that were previously generated from samples of primary cervical cancer were re-analyzed. The samples were grouped based on their histopathology, comparing SCC to ADC. Each tumor in the study was confirmed to be high risk human papilloma virus (hrHPV) positive. A total of 21 cervical cancer samples were included, with 11 cases of SCC and 10 of ADC. Data analysis revealed a total of 26 differentially expressed genes, with 19 genes being overexpressed in SCC compared to ADC (Benjamini–Hochberg (BH)-adjusted p-value < 0.05). Importantly, the immune checkpoint markers CD274 and CTLA4 demonstrated significantly higher expression in SCC compared to ADC. In addition, SCC showed a higher infiltration of immune cells, including B and T cells, and cytotoxic cells. Higher activation of a variety of pathways was found in SCC samples including cytotoxicity, interferon signaling, metabolic stress, lymphoid compartment, hypoxia, PI3k-AKT, hedgehog signaling and Notch signaling pathways. Our findings show distinctive gene expression patterns, signaling pathway activations, and trends in immune cell infiltration between SCC and ADC in cervical cancer. This study underscores the heterogeneity within primary cervical cancer, emphasizing the potential benefits of subdividing these tumours based on histological and molecular differences.

Triage methods for cervical cancer detection show moderate accuracy and present considerable false-negative and false-positive result rates. A complementary diagnostic parameter could help improve the accuracy of identifying patients who need treatment. A pilot study was performed using a targeted proteomics approach with opportunistic ThinPrep samples obtained from women collected at the hospital’s outpatient clinic to determine the concentration levels of minichromosome maintenance-3 (MCM3) and envoplakin (EVPL) proteins. Forty samples with ‘negative for intraepithelial lesion or malignancy’ (NILM), 21 samples with ‘atypical squamous cells of undetermined significance’ (ASC-US), and 33 samples with ‘low-grade squamous intraepithelial lesion and worse’ (≥LSIL) were analyzed, using cytology and the patients’ histology reports. Highly accurate concordance was obtained for gold-standard-confirmed samples, demonstrating that the MCM3/EVPL ratio can discriminate between non-dysplastic and dysplastic samples. On that account, we propose that MCM3 and EVPL are promising candidate protein biomarkers for population-based cervical cancer screening.

Objective: Cervical cancer and its precursor are labeled as HIV indicator conditions, justifying the recommendation of HIV testing for all patients. This study aimed to assess the acceptance and patients' and gynecologists' perceptions on HIV testing in patients with cervical dysplasia at the colposcopy outpatient clinic in hospitals in the Netherlands.Methods: A cross‐sectional study was conducted between May 2021 and February 2023 to implement point‐of‐care HIV testing in five hospitals in the Rotterdam region, the Netherlands. We included patients aged 18 years and older, without documented HIV, presenting with cervical dysplasia. The primary outcome was the HIV test acceptance rate. Secondary outcomes were the HIV positivity rate and the association between dysplasia severity and test acceptance. We also assessed patients' and gynecologists' perspectives on this testing strategy with a questionnaire.Results: Of 563 patients, 523 accepted HIV testing, resulting in a test acceptance rate of 92.9% (95% confidence interval, 90.45%–94.88%). Testing rates were consistent among participating hospitals (91.6%–100.0%) None of the patients tested positive for HIV. Severity of dysplasia was not associated with test acceptance (P = 0.768). Most patients (n = 500, 96.0%) reported good or higher experience with this testing strategy. The main barriers for gynecologists to offer HIV testing were lack of time (n = 22, 33.9%) and fear to offend a patient (n = 7, 12.5%). Less than half (n = 19, 39.3%) of gynecologists believed an HIV test should be offered to all patients with cervical dysplasia.Conclusion: Point‐of‐care HIV testing at colposcopy outpatient clinics was well accepted by patients. The data indicate that the primary barrier lies with the physicians to offer such testing.

We conducted a comparative spatial protein analysis of primary cervical tumors and lymph nodes (LNs) with and without metastasis to uncover immune signatures linked to metastatic progression. Spatial protein profiling was conducted using the GeoMx Digital Spatial Profiler using 14 primary cervical cancer (CC) specimens (seven patients with LN metastasis and seven without). In primary tumors, CD8 and CD25 were significantly upregulated in non-metastatic compared to metastatic cases. In LNs, ICOS expression was significantly higher in tumor-free LNs of patients without metastasis compared to tumor-free LNs of patients with metastasis. In metastatic LNs, the significant findings varied depending on the distance between tumor cells and the germinal centers. The distant GCs exhibited significantly higher expression of CD11c, CD68, Ki-67, and PD-1 compared to GCs of non-metastatic LNs. Overall, non-metastatic tumors and LNs exhibit more robust immune activation, whereas metastatic sites demonstrate clear immune suppression and structural disruption. These findings suggest that early immune dysregulation may contribute to metastatic spread and could inform future therapeutic strategies.

Ovarian cancer (OC) is typically treated with cytoreductive surgery (CRS). Hyperspectral imaging (HSI) is an emerging non-invasive, label-free technique that enables whole-area scanning, making it a promising tool for real-time tumour recognition. However, developing tumour recognition algorithms requires a foundational understanding of spectral variability in normal tissues. This study focusses on the in vivo spectral profiles of key abdominal and pelvic organs encountered during CRS, including the uterus, ovaries, intestines, mesentery, omentum, peritoneum, and fallopian tubes, and evaluates the potential for organ recognition using HSI data. Intraoperative HSI data were from healthy patients. Two machine learning models, a support vector machine (SVM) and a 3D convolutional neural network (3DCNN), were trained to classify the organs based on their spectral signatures. In total, 15 patients were included in the dataset. The 3DCNN slightly outperformed the SVM in terms of the average accuracy (0.889 vs. 0.878), sensitivity (0.648 vs. 0.604), specificity (0.936 vs. 0.930), and Dice Similarity Coefficient (0.595 vs. 0.569). This study demonstrates the feasibility of using HSI for organ differentiation in the clinical setting, although in some cases separability remains a challenge, especially when organs have similar spectra. This is a critical step towards a generalizable in vivo abdominal tumour recognition algorithm, by carefully investigating spectral fingerprints of abdominal tissues.