BACKGROUND:The methylation of paired boxed gene 1 (PAX1) and zinc finger protein 582 (ZNF582) has been proved to have good diagnostic efficacy to detect cervical precancerous lesions in Asian populations, but their performance in Dutch populations is still unclear. We assessed PAX1 and ZNF582 methylation in both Dutch and Chinese cohorts. METHODS:Cervical scrapings were evaluated from 2 independent groups. The Dutch cohort (population-based screening) provided 190 abnormal cytology scrapings, and 138 cases tested positive for high-risk human papillomavirus (hrHPV). The hospital-based Chinese cohort included 246 specimens (205 abnormal cytology and 227 hrHPV+). Quantitative methylation-specific PCR was employed to analyze methylation. RESULTS:PAX1 and ZNF582 methylation showed high specificity of 91% to 97% with moderate sensitivity of 43% to 61% for cervical intraepithelial neoplasia (CIN) 2 or worse (CIN2+) in abnormal cytology and hrHPV+ samples in both cohorts. Sensitivity for CIN3+ increased to 55% to 86% but with lower specificity (85%-93%). For detecting CIN2+, PAX1/ZNF582 methylation performed similarly in Dutch and Chinese cohorts. However, for CIN3+, the Chinese cohort showed significantly higher sensitivity, with comparable specificity. We also evaluated combining these genes with our established markers (C13ORF18, JAM3, and ZSCAN1). The best diagnostic performance was achieved by combining PAX1 or ZNF582 with JAM3 in the Dutch cohort. CONCLUSIONS:In conclusion, regardless of race or geography, methylation of PAX1 or ZNF582 is highly specific for detecting CIN2+/CIN3+ lesions in hrHPV+ or cytologically abnormal scrapings. Triage testing using these markers might significantly reduce immediate colposcopies. IMPACT:PAX1/ZNF582 methylation triage for cytology+/hrHPV+ scrapings may reduce immediate colposcopies.

Background For patients with early-stage vulvar squamous cell carcinoma (VSCC), the indication for groin surgery currently depends mainly on the depth of invasion (DOI). In 2021, the International Federation of Gynaecology and Obstretrics (FIGO) revised the DOI measurement method in its updated guidelines, addressing the clinical need to improve the prognostic distinction between stage IA and IB. Our aim was to investigate the extent to which stage migration occurs when the FIGO 2021 method is applied, and whether lymph node involvement may be missed due to the omission of groin exploration in FIGO 2021 stage IA disease. Method Patients who participated in GROINSS-V I or II and treated in the University Medical Center Groningen (UMCG) were included (n= 245). All patients had primary VSCC <4cm, with a DOI >1mm and non-suspicious groin nodes. Tumor thickness, the FIGO 2009 method, and the FIGO 2021 method were annotated on each slide.Findings - In total, 212/245 (86.5%) patients were available for analysis. Twenty-four (24/212,11.3%) had a DOI >1mm according to the FIGO 2009 method and ≤1 mm according to the FIGO 2021 method, which would have resulted in a stage shift to IA. Of the downstaged patients, four had a SN metastasis and one had an isolated groin recurrence after a negative SN (5/24, 20.8% [95% CI: 9.2% - 40.5%]). Interpretation Our results indicate that, although the implementation of the FIGO 2021 method will result in less SN procedures and thereby less treatment related morbidity, this comes at the cost of more missed metastases in microinvasive disease, risking worse outcomes for these patients. We urgently request the FIGO committee to reconsider the recommended DOI method.

Interest in understanding the high chemoresistance and poor prognosis of advanced ovarian clear cell carcinoma (OCCC) is rising. Patient-derived xenografts (PDX) are widely used in vivo models because of their supposedly accurate morphologic and (epi)genetic representation of patient tumors. Here, we established five subcutaneous OCCC PDXs. The PDX.F1 engraftment success rate was over 30% with similar latency time and growth speed of PDX.F2. ARID1A, PTEN, ATM, BRCA1 and PIK3CA mutations were found in matched tumors and PDXs. ARID1A protein loss was further verified by immunohistochemical staining. Cyclophilin A staining depicted the replacement of human stroma by mouse stroma in PDX.F2, while PAS/PAS-D staining confirmed cellular glycogen accumulation in OCCC tumors and PDXs. SNP array and Infinium MethylationEPIC BeadChip array data analysis demonstrated the copy number alterations and DNA methylation signatures of genome-wide and tumor-driver genes in PDXs generally resembled their patients’ tumors. Promoter CpG islands of a small number of genes, enriched in PRC2/histone methylation related gene-sets, gained methylation (△β-value > 0.4) in PDXs vs patient tumors. In conclusion, the high phenotypic and molecular similarity allows the established PDXs to serve as potential preclinical models for future translational research of OCCC.

Early-stage vulvar cancer is managed by a local excision of the primary tumor and, if indicated, a sentinel node (SN) biopsy to assess the need for further groin treatment. With the SN procedure, many patients can be treated less radically and will experience less complications and morbidity compared with an inguinofemoral lymphadenectomy (IFL). Still, the SN procedure can be further optimized. Different tracers for detecting the SN are being investigated, aiming to optimize detection rates and decrease the burden of the procedure and short-term complications. Until now, no standardized protocols exist for the pathologic workup of the SN, possibly leading to discrepancies in detection of metastases between institutes using different methods. New techniques, such as one-step nucleic amplification, seem to have potential in accurately detecting metastases in other cancers, but have not yet been investigated in vulvar squamous cell carcinoma (VSCC). Furthermore, several studies have investigated the possibility to broaden the indications for the SN procedure, such as its use in recurrent disease, larger tumors, or multifocal tumors. Although these studies show encouraging results, cohorts are small and further studies are needed. Prospective studies are currently investigating these subgroups. Lastly, several studies investigated optimization of groin treatment of patients with a metastatic SN. Inguinofemoral radiotherapy is a good alternative to IFL in patients with micrometastases in the SN, with comparable efficacy and less treatment-related morbidity. Reduction of the radicality of groin treatment is also possible in other ways, such as omitting contralateral IFL in patients with lateralized tumors and a unilateral metastatic SN. In conclusion, the SN procedure is an established procedure in early-stage VSCC, although optimization of the technique, pathologic workup, indications, and treatment in the setting of metastatic disease are the subject of ongoing research.

BackgroundAs part of its mission to improve the quality of care for women with gynecological cancers across Europe, the European Society of Gynaecological Oncology (ESGO) first published in 2017 evidence-based guidelines for the management of patients with vulvar cancer. ObjectiveTo update the ESGO guidelines based on the new evidence addressing the management of vulvar cancer and to cover new topics in order to provide comprehensive guidelines on all relevant issues of diagnosis and treatment of vulvar cancer. MethodsThe ESGO Council nominated an international development group comprised of practicing clinicians who provide care to vulvar cancer patients and have demonstrated leadership through their expertize in clinical care and research, national and international engagement and profile as well as dedication to the topics addressed to serve on the expert panel (18 experts across Europe). To ensure that the statements were evidence-based, new data identified from a systematic search were reviewed and critically appraised. In the absence of any clear scientific evidence, judgment was based on the professional experience and consensus of the international development group. Prior to publication, the guidelines were reviewed by 206 international practitioners in cancer care delivery and patient representatives. ResultsThe updated guidelines cover comprehensively diagnosis and referral, staging, pathology, pre-operative investigations, surgical management (local treatment, groin treatment, sentinel lymph node procedure, reconstructive surgery), (chemo)radiotherapy, systemic treatment, treatment of recurrent disease (vulvar, inguinal, pelvic, and distant recurrences), and follow-up. Management algorithms are also defined.

Supplementary Figure 5 | Clustering (n=106) and survival analysis (n=70) on kinome mutations and CNA. (A) Disease-specific survival of patients (n=70) grouped on ARID1A status, (B) PIK3CA status and (C) ARID1A + PIK3CA status. (D) Consensus clustering, with maximum group count set to 10 and clustering optimization with 1000 repetitions maximum, shows adding of tumors (n=106) as consensus index (horizontally) and empirical cumulative distribution (vertically) for 1-10 clusters. (E) Area under the curve plot of decrease in friction from 1-10 clusters. (F) Heatmap distribution of tumors in 8 clusters. (G) Disease-specific survival of patients (n=70) in 8 clusters, (H) cluster 3 vs. other clusters. (I) Disease-specific survival in advanced stage OCCC patients (FIGO 2C-4), Log rank (Mantel-Cox) was used for statistical analysis. (J) Nonsynonymous mutation distribution in genes involved in the frequently mutated PI3K/AKT/mTOR (blue) and MAPK pathway (yellow), ERBB family of receptor tyrosine kinases (green) and DNA repair pathway (red) as well as ARID1A and PALB2 are shown with OncoPrint. CNA in each mutated gene are added. The 106 OCCC tumors that were both kinome sequenced and SNP arrayed are shown on the horizontal axis grouped on tumor clusters and ordered on total event frequency in the subsequently altered pathways. (K) Oncoprint from advanced stage patients in cluster 3 vs. other clusters, shown on the horizontal axis grouped on tumor clusters and ordered on total event frequency in the subsequent altered pathways, ordered as aforementioned.

Supplementary Figure 4 | Whole genome CNA heatmap. (A) Genome-wide CNA heatmap profiles of all 108 OCCC tumors (above), 63 ARID1A wildtype tumors (middle) and 45 ARID1A mutant tumors (below). (B) Significant CNA plot of only ARID1A wildtype tumors (n=63) and (C) only ARID1A mutant tumors (n=45) as determined by GISTIC analysis. All kinases and cancer-related genes from the kinome sequencing gene panel that were focally significantly amplified (red) or deleted (blue) are indicated along the chromosomes vertically. Chromosomal location and total amount of tumors harboring the event are annotated with each gene name. False-discovery rate (FDR) 0.05 threshold, indicated by the green line, and G-score are shown along the horizontal axis.

Supplementary Data from Expression of Epidermal Growth Factor Receptor (EGFR) and Activated EGFR Predict Poor Response to (Chemo)radiation and Survival in Cervical Cancer

Supplementary Table S2 from Involvement of the TGF-β and β-Catenin Pathways in Pelvic Lymph Node Metastasis in Early-Stage Cervical Cancer

Supplementary Table 5 from Genome-Wide Promoter Analysis Uncovers Portions of the Cancer Methylome

Supplementary Table 3 | Binomial testing of kinome sequencing variants.

Supplementary Table 2 from Genome-Wide Promoter Analysis Uncovers Portions of the Cancer Methylome

Supplementary Table 1 | Gene set for kinome sequencing.

Supplementary Table 8 from Genome-Wide Promoter Analysis Uncovers Portions of the Cancer Methylome

Supplementary Table 1 from Genome-Wide Promoter Analysis Uncovers Portions of the Cancer Methylome

Supplementary Figures S1-S2 from Involvement of the TGF-β and β-Catenin Pathways in Pelvic Lymph Node Metastasis in Early-Stage Cervical Cancer

PDF file - 20K, Schematic representation of the study design