APMIS

HERC5: a comprehensive in silico analysis of its diagnostic, prognostic, and therapeutic potential in cancer

HERC5, a vital protein in the HERC family, plays crucial roles in immune response, cancer progression, and antiviral defense. This bioinformatic study comprehensively assessed HERC5's significance across various malignancies by analyzing its gene expression, immune and molecular subtype expressions, target proteins, biological functions, and prognostic and diagnostic values in pan‐cancer. We further examined its correlation with clinical features, co‐expressed and differentially expressed genes, and prognosis in clinical subgroups, focusing on endometrial cancer (UCEC). Our findings showed that HERC5 RNA is expressed at low levels in most cancers and significantly differs across immune and molecular subtypes. HERC5 accurately predicts cancer and correlates with most cancer prognoses. In UCEC, HERC5 was significantly associated with age, hormonal status, clinical stage, treatment status, and metastasis. Elevated HERC5 expression was linked to worse progression‐free interval, disease‐specific survival, and overall survival in UCEC, particularly in diverse clinical subgroups. Significant differences in HERC5 expression were also observed in various human cancer cell line validations. In summary, HERC5 may be a critical biomarker for pan‐cancer prognosis, progression, and diagnosis, as well as a promising new target for cancer therapy.

Dishevelled family proteins in serous ovarian carcinomas: a clinicopathologic and molecular study

Dishevelled family proteins (DVL1, DVL2, and DVL3) are cytoplasmic mediators involved in canonical and non‐canonical Wnt signaling that are important for embryonic development. Since Wnt signaling promotes cell proliferation and invasion, its increased activation is associated with cancer development as well. To get deeper insight into the behavior of Dishevelled proteins in cancer, we studied their expression in serous ovarian carcinomas [both low‐ (LGSC) and high‐grade (HGSC)], and HGSC cell lines OVCAR5, OVCAR8, and OVSAHO. DVL protein expression in serous ovarian carcinomas tissues was analyzed using immunohistochemistry, while DVL protein and mRNA expressions in HGSC cell lines were analyzed using Western blot and quantitative real‐time PCR. DVL1 protein expression was significantly higher in LGSC compared with normal ovarian tissue, while DVL3 was overexpressed in both LGSC and HGSC. DVL2 and DVL3 protein expression was higher in HGSC cell lines when compared with normal control cell line FNE1, while DVL1, DVL2, and DVL3 mRNA expression was significantly increased only in OVSAHO cell line. Survival analysis revealed no significant impact of DVL proteins on patients’ outcome. Our data show an active involvement of Dishevelled family proteins in serous ovarian carcinomas. Further studies should confirm the clinical relevance of these observations.

Immunotyping in tubo‐ovarian high‐grade serous carcinoma by PD‐L1 and CD8+ T‐lymphocytes predicts disease‐free survival

AbstractPD‐L1 immune checkpoint inhibitor expression was evaluated in high‐grade serous carcinoma (HGSC) ovary in the context of the overall immune landscape to determine its prognostic value. Consecutive cases of HGSC, 50 who underwent upfront surgery followed by adjuvant chemotherapy (HGSC‐U) and 50 who underwent neoadjuvant chemotherapy (NACT) followed by interval debulking surgery (HGSC‐PC) were selected. In HGSC‐PC cases, the pre‐NACT ascitic fluid cell blocks were included. Tumor‐infiltrating lymphocytes (TILs) were scored, hotspots chosen for tissue microarray construction and immunohistochemistry performed and scored for CD4 and CD8 lymphocyte subsets, CD68+ tumor‐associated macrophages (TAMs), PD‐1 and PD‐L1 expression. HGSC‐post‐chemotherapy showed increased TILs, predominantly CD8+T‐lymphocytes, compared to HGSC‐U. HGSC showed PD‐L1 expression on tumor cells and/or TAMs in 60% cases with a linear correlation to CD4+, CD8+ TIL levels. Concordant PD‐L1 expression was seen in matched pre‐ and post‐NACT tumor cells. HGSC‐PC showed higher expression of PD‐L1. There was no association of PD‐L1 cumulative proportion score or tumor cell score with outcome. Taking a cutoff for PD‐L1 CPS at 10%, immunotype I (PD‐L1+/CD‐8+), corresponding to tumors with adaptive immune evasion, showed worst disease‐free survival compared to all other immunotypes (p = 0.03) and was more significant (p = 0.01) when compared to immunotype III (PD‐L1+/CD8−). Immunotyping based on PD‐L1/CD8+ expression correlates to prognosis and outcome.

Optimization of Liquid‐Based Cytology Block Preparation Using Modified Adhesive Centrifugal Sedimentation for Improved Diagnostic Accuracy in Cervical Lesions

ABSTRACT Liquid‐Based Cytology (LBC) is widely used for cervical cancer screening, which significantly improves cell dispersion and morphological preservation compared to cytology smears, but its detection rate is still limited, and the LBC block further improves on its technical deficiencies. This study optimized LBC block preparation by evaluating five methods using 90 cervical samples. Metrics included visual assessment, hematoxylin and eosin (H&E) staining, Immunohistochemistry (IHC) performance, and production efficiency. The modified adhesive centrifugal sedimentation method demonstrated superior outcomes, yielding well‐preserved cell clusters with enhanced H&E contrast (reduced impurities) and precise IHC localization. Its preparation success rate reached 92.9%. Validation with 114 cervical cases showed cell blocks achieved 87.5% sensitivity and 96.9% specificity compared to biopsy histology, outperforming standard LBC. The method's simplicity, cost‐effectiveness, and cell preservation advantages support its utility in primary settings for cervical lesion screening. Findings highlight LBC blocks as a reliable adjunct to cytology, improving diagnostic precision while maintaining workflow feasibility.

Large‐Scale Study Comparing Analytical and Diagnostic Quality of Three HPV Self‐Sampling Devices for At‐Home Cervical Cancer Screening

ABSTRACT Elimination of cervical cancer requires broad access to and participation in cervical screening. Self‐sampling has emerged as a robust technology that simplifies access; however, few self‐sampling devices are independently validated regarding sample quality. This study compares the quality of three self‐sampling devices: Evalyn (Rovers), currently used in Denmark, the FLOQSwab (Copan) and the modified FLOQSwab: SensiGrip. Women residing in the Capital Region of Denmark, accepting screening by self‐sampling, were offered a kit containing the following combinations of devices: (1) Evalyn, FLOQSwab, (2) Evalyn, SensiGrip or (3) FLOQSwab, SensiGrip. Returned kits were analyzed using the validated BD Onclarity HPV assay on the COR instrument (BD). A total of 1677 women participated. Sample quality was similar across devices, also when stratifying into age groups. Two hundred thirty‐two women (13.9%) tested positive for one or more oncogenic HPV types. Pairwise concordance analysis for each group showed an overall agreement between 93.5% and 95%. Analytical and diagnostic performance of samples collected by the three self‐sampling devices resulted in similar quality and HPV detection. Hence the sample quality is not a determinant in the choice of sampling device and focus on other factors such as cost, women's preference or size and weight can take precedence.

Cytohistological findings in HPV‐negative cases from HPV primary screening programme: quality assurance study

To evaluate the risk of false HPV‐negative results and possible related morphological abnormalities in HPV primary cervical cancer screening. Out of 53,661 HPV‐negative cases, 5469 (10.2%) randomly selected cytology slides were evaluated as a part of the quality assurance protocol. The Bethesda category negative for intraepithelial lesion or malignancy (NILM) in the HPV‐negative cases given was present in 95.4%. Due to cytology other than NILM, 0.4% of cases were referred to colposcopy and 4.2% to the follow‐up in one year. In the follow‐up HPV negativity and NILM cytology was present in 88.3% of attended women. Cases other than HPV negative and NILM were referred to colposcopy. One biopsy‐proven histological HSIL was found in the first round and one in follow‐up screening. More comprehensive genotyping of HSIL cases revealed genotypes 69 and 11. Only two HPV test negative cases with histological HSIL were revealed forming 0.04% of quality control group. In both cases, HPV genotype not included in screening tests was found. According to the results, the primary HPV test with cytology triage is an efficient and specific method for cervical cancer screening despite of the fact that some non‐high‐risk genotypes are missed.

Are we ready for translational research based on material and data from the Danish CancerBiobank and can we gain new knowledge from biobank registration?

Bio‐and GenomeBank, Denmark (RBGB) is a nationwide infra‐structure. Danish CancerBiobank (DCB) is a biobank in RBGB. The aim is to describe the degree of biological material collected and stored in DCB for patients diagnosed with primary ovarian cancer registered in The Danish Gynecologic Cancer Database (DGCD). Furthermore, to investigate the concordance between predicted organ of disease registered in RBGB at time of sampling (presumed diagnosis) with final diagnosis for patient. Data extraction from DGCD and DCB. Biological materials are present for 1.347 (62%) of 2.172 patients with primary ovarian cancer (OC). The median age of OC patients were 68 years (range: 18–90 years). Median age of patients with biological material in DCB was 67 years and for patients without biological material in DCB 69 years (p ≤ 0.0001). The histological subtypes for the 1347 OC patients with biological material were 911 (68%) serous adenocarcinoma, 97 (7%) endometrioid adenocarcinoma, 80 (6%) mucinous adenocarcinoma, 58 (4%) clear cell carcinoma, and for 201 (15%) no information were registered. For 327 patients (24%), the presumed diagnosis was hematological with a final diagnosis of OC. Using clinical data and biological material including pre‐analytical data regarding the biological material the possibility for translational research is optimal. Furthermore, information registered through daily working procedures may propose the need for additional biomarkers to aid clinicians to stratify patients to treatment in correct fast‐track packages.

No Bacterial Biomass Detected in Tissue From Patients With Ovarian Cancer and Serous Tubal Intraepithelial Carcinomas Using 16S rDNA Sequencing

The ovarian oncobiome is subject to increasing scientific focus, but a potential link between bacterial dysbiosis and ovarian carcinogenesis remains controversial. Our primary aim was to characterize the bacterial microbiota in epithelial ovarian cancer samples. Secondarily, we aimed to compare results from the bacterial microbiota in formalin‐fixed, paraffin‐embedded ovarian tissue samples from 194 patients with epithelial ovarian cancer, fallopian tube tissue samples from 16 patients with serous tubal intraepithelial carcinomas and in benign fallopian tube tissue samples from 25 patients. Bacterial abundance was investigated by means of 16S rDNA Next Generation Sequencing. The 16S rDNA sequencing run produced a very low number of bacterial reads. Only seven samples displayed bacterial reads above 5000. Validation of detected bacterial reads by qPCR was negative and indicative of results being background amplification. Our results indicate no amount of bacterial biomass in the ovarian cancer, benign fallopian tube and in the samples with serous tubal intraepithelial carcinomas. The findings underline the importance of validating results from bacterial microbiota studies with additional technical assays before any conclusion may be drawn.

Vitamin A deficiency in K14E7HPV expressing transgenic mice facilitates the formation of malignant cervical lesions

AbstractInfection with high‐risk human papillomavirus (HR‐HPV) is the main cause of cervical cancer (CC), but viral infection alone does not guarantee the development of this malignancy. Indeed, deficiencies of dietary micronutrients could favor cervical cancer development in individuals that harbor HR‐HPV infections. The status of retinoid levels, natural and synthetic derivatives of vitamin A, is important in maintaining cellular differentiation of the cervical epithelium. Moreover, many studies show a link between deficient intake of retinoids or alteration of the retinoid receptors and CC development. In spite of this, the effect of vitamin A deficiency (VAD) in presence of HR‐HPV oncoproteins on cervical carcinogenesis in vivo has not been reported. Transgenic mice expressing E6 or E7 oncoproteins (K14E6 or K14E7 mice, respectively) were used to evaluate the possible role of VAD in the development of malignant cervical lesions. The survival of the mice in VAD condition was studied, and histopathological analysis and immunohistochemical detection of molecular cancer markers such as the tumor suppressor retinoic acid receptor beta (RARβ), proliferating cell nuclear antigen (PCNA), cleaved caspase 3, and the tumor suppressor protein p16INK4A (inhibitor of CDK4) were performed. Our results show that K14E6/VAD mice showed moderate cervical dysplasia; notably, K14E7/VAD mice developed severe cervical dysplasia and cervical in situ carcinoma at an early age. VAD synergizes with HPV16E7 oncoprotein expression favoring cervical carcinogenesis in vivo.

Immunohistochemical expression of SATB2 and PAX8 in differentiating primary from metastatic ovarian mucinous neoplasms

Accurate stratification of an ovarian mucinous neoplasm as primary or secondary is always challenging as they show overlapping histomorphological and immunohistochemical features. Immunohistochemical staining for SATB2 and PAX8 was performed on 80 cases of mucinous ovarian neoplasms subdivided into 53 primary [25 primary ovarian mucinous carcinomas (POMCs) and 28 mucinous borderline tumors (MBTs)] and 27 secondary (12 of colonic origin, 7 of appendiceal origin, and 8 of gastric origin). Expression was correlated with different clinicopathologic parameters. PAX8‐positive immunostaining was detected in 38 out of 53 cases (71.69%) of primary ovarian mucinous neoplasms (POMNs) with null positivity in the secondary ovarian mucinous tumors (0/27). SATB2‐positive expression was detected in 16 out of 27 cases (59.26%) of the secondary ovarian mucinous tumors. None of the studied POMNs showed any positive immunostaining for SATB2 (0/53). A profile of SATB2−/PAX8+ and SATB2+/PAX8− can be used to differentiate POMNCs from secondary ovarian mucinous tumors of GI origin, respectively, with 100% specificity. PAX8 expression is associated with some clinicopathologic parameters providing the basis for the possible usage of PAX8 as prognostic marker.

Fumarate hydratase–deficient renal cell carcinoma: an oncology care institutional experience

Renal cell carcinoma (RCC) accounts for 2% of all cancer cases worldwide, and majority are sporadic. The latest World Health Organization (WHO) classification of renal cell tumors (fifth edition, 2022) has molecularly defined renal tumor entities, which includes fumarate hydratase (FH)–deficient RCC. FH‐deficient RCC is an aggressive carcinoma caused by pathogenic alterations in FH gene, seen in 15% of patients with hereditary leiomyomatosis and renal cell cancer syndrome (HLRCC) syndrome. These tumors occur more frequently at a younger age and present at an advanced stage, carrying a dismal prognosis. We report a series of 10 cases of FH‐deficient RCC. The mean age was 49.8 years, and all cases presented in advanced stages (III and IV). Morphologically, the cases had varied architectural patterns with characteristic eosinophilic macronucleoli and perinucleolar halo. On immunohistochemistry (IHC), all showed diffuse nucleo‐cytoplasmic expression of S‐(2‐succino)‐cysteine (2‐SC), with loss of FH in seven cases. FH‐deficient RCCs are aggressive neoplasms and can be diagnosed using specific IHC markers (FH and 2‐SC). These patients should undergo germline testing for FH gene mutation, genetic counseling, and surveillance of family members.

Modulation with anti‐Oma87 antibodies of cytotoxicity, adherence, and internalization of Acinetobacter baumannii in human cervical carcinoma epithelial cells

BamA, an Omp85 superfamily member, is universally conserved and essential for cell viability. Using anti‐Oma87 antibodies, we focus on understanding the effect of Oma87 of Acinetobacter baumannii on pathogenicity. Oma87 was expressed, purified, and used to induce anti‐Oma87 antibodies in BALB/c mice. Acute toxicity of the protein was evaluated in mice. HeLa cells were infected with both live and killed A. baumannii 19606 and a clinical isolate. The effects of anti‐Oma87 sera on A. baumannii adherence, internalization, and proliferation in HeLa cells were studied. The roles of microfilaments and microtubules in A. baumannii invasion were demonstrated by Actin disruption. Reduced bacterial population and biofilm formation were noted. The ability of A. baumannii to provoke autophagy through Oma87 induction leads to incomplete autophagy and potentially facilitates bacterial replication. Actin‐mediated uptake, attachment, and invasion demonstrated A. baumannii survival and multiplication within vacuoles in the host cell. The findings underscore the potential of Oma87 as a therapeutic intervention target in infections caused by A. baumannii. This comprehensive analysis contributes valuable information for understanding the virulence mechanisms of A. baumannii, potentially guiding future strategies to combat infections caused by this pathogen.

The role of Pap smear in the diagnostics of endocervical adenocarcinoma

In the high‐income countries, the amount of cervical adenocarcinomas is on the rise. The pap smear sampling has a low sensitivity and a low specificity for endocervical malignancies, and there are only a few cytomorphological features, that are specifically associated with glandular atypia. In this study, 298 pap smears of 60 patients with endocervical adenocarcinoma or adenocarcinoma in situ (AIS) and 30 patients with high‐grade intraepithelial lesion (HSIL) in histology were reviewed. The pap smear type (screening/clinical), the HPV status and the time from sampling to the histological confirmation of diagnosis were recorded for each case. Despite that no cytomorphological features could be associated with adenocarcinoma statistically, 70% of the pap smears were initially correctly diagnosed as an endocervical glandular lesion. Palisading cell borders, nuclear pleomorphism and the lack of single atypical cells present simultaneously were found to be associated with adenocarcinoma and AIS with the corresponding ORs of 5.89 (95% CI 1.96–17.70), 3.71 (95% CI 1.14–12.02) and 10.76 (95% CI 1.20–59.50). This combination of features was seen in smears taken up to 5 years before the histological diagnosis. Of all our screening samples, 10.9% were HPV‐positive. There were no HPV‐negative samples among patients with adenocarcinoma.

SATB2 is Rarely Expressed in Endometrial or Endocervical Carcinoma

ABSTRACT Special AT‐rich sequence‐binding protein 2 (SATB2) can distinguish primary ovarian mucinous tumors from ovarian metastases of colorectal or appendiceal tumors. However, its expression in other gynecological localizations remains underexplored. Whole‐tumor sections of 376 endometrial and 27 endocervical carcinomas were examined for SATB2 expression. Among endometrial carcinomas, we found 10 cases (2.7%) expressing SATB2, all of which were endometrioid adenocarcinomas. In 8 of them, expression concerned only the morules. Only two cases expressed SATB2 in the glandular component, indicating 0.53% positivity in this cancer group. No stromal expression was detected. One case (3.8%) of endocervical adenocarcinoma focally expressed SATB2. SATB2 expression can be reliably used in the differential diagnosis of primary endometrial endometrioid and serous carcinomas as well as primary endocervical adenocarcinomas because nearly all of these tumors are SATB2‐negative. SATB2 positivity in endometrial carcinomas is almost always restricted to morules within endometrioid adenocarcinomas.

Vaginal microecology and its role in human papillomavirus infection and human papillomavirus associated cervical lesions

The vaginal microecology comprises the vaginal microbiome, immune microenvironment, vaginal anatomy, and the cervicovaginal fluid, which is rich in metabolites, enzymes, and cytokines. Investigating its role in the female reproductive system holds paramount significance. The advent of next‐generation sequencing enabled a more profound investigation into the structure of the vaginal microbial community in relation to the female reproductive system. Human papillomavirus infection is prevalent among women of reproductive age, and persistent oncogenic HPV infection is widely recognized as a factor associated with cervical cancer. Extensive previous research has demonstrated that dysbiosis of vaginal microbiota characterized by a reduction in Lactobacillus species, heightens susceptivity to HPV infection, consequently contributing to persistent HPV infection and the progression of cervical lesion. Likewise, HPV infection can exacerbate dysbiosis. This review aims to provide a comprehensive summary of current literatures and to elucidate potential mechanisms underlying the interaction between vaginal microecology and HPV infection, with the intention of offering valuable insights for future clinical interventions.

Circulating miR‐15b, miR‐34a and miR‐218 as promising novel early low‐invasive biomarkers of cervical carcinogenesis

Circulating biological markers, such as miRNAs, hold the greatest possibilities to complement tissue biopsy and clinical diagnostic tests. The objective of this study was to evaluate the relative abundance of three circulating miRNAs in serum from 17 HPV16‐positive patients with early cervical lesions known as Low‐Grade Squamous Intraepithelial Lesions (LSILs). The expression of circulating microRNAs miR‐15b, miR‐34a and miR‐218 in patients with LSILs was compared to 23 HPV‐negative individuals showing normal cervical epithelium (healthy women) and 23 Squamous Cell Carcinoma (SCC) samples. The expression levels of miR‐15b remained unchanged while those of miRNAs 34a and 218 were relatively high in serum obtained from LSIL patients in comparison with healthy women (results were statistically significant with a p of < 0.01 or < 0.001). According to previous findings, miR‐15b was overexpressed and miRNAs 34a and 218 were underexpressed in serum from SCC patients. Additionally, the mRNA expression levels of some selected gene targets were determined [Cyclin D1 (CCND1), Cyclin E1 (CCNE1), B‐cell lymphoma 2 (Bcl‐2) and MutS homolog 2 (MSH‐2)]. All serum results correlated with tissue samples from the same patients. We propose that circulating microRNAs can be valuable as molecular markers for the early follow‐up of cervical carcinogenesis risk.

Undifferentiated–dedifferentiated endometrial carcinoma; the reappearance of an old friend with insights into the new data

Endometrial carcinoma is a common malignancy in women and shows increasing incidence. Except for its two main pathogenetic types I and II, the continuing evolution on molecular genetics have led to a new classification system (TCGA), that includes four main molecular subtypes: (i) POLE‐mutant (ultramutated), (ii) hypermutated (MSI), (iii) copy‐number low/MSS (p53wt) and (iv) copy‐number high/serous‐like (p53mut). The undifferentiated and dedifferentiated endometrial carcinomas are rare and clinically aggressive variants, comprising about 10% of the high‐grade endometrial carcinomas and 2% of the endometrial carcinomas in general. Until recently, they were under‐recognized and not fully described morphologically and immunohistochemically/molecularly. Their recognition diagnostically is crucial because of their poor prognosis; approximately 40% of patients with these subtypes will die within 0.5–20 months after diagnosis, so additional therapeutic strategies are important for an effective management. Because of their rarity, the responsiveness to other than conventional treatment, such as immunotherapy, has not been sufficiently investigated yet. The aim of this review is to provide an update on the knowledge about these two uncommon subtypes according to the current literature.