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ObjectivesVulvar squamous cell carcinoma is subclassified into three prognostically relevant groups: (i) human papillomavirus (HPV) associated, (ii) HPV independent p53 abnormal (mutant pattern), and (iii) HPV independent p53 wild type. Immunohistochemistry for p16 and p53 serve as surrogates for HPV viral integration andTP53mutational status. We assessed the reproducibility of the subclassification based on p16 and p53 immunohistochemistry and evaluated the prognostic significance of vulvar squamous cell carcinoma molecular subgroups in a patient cohort treated by vulvar field resection surgery.MethodsIn this retrospective cohort study, 68 cases treated by vulvar field resection were identified from the Leipzig School of Radical Pelvic Surgery. Immunohistochemistry for p16 and p53 was performed at three different institutions and evaluated independently by seven pathologists and two trainees. Tumors were classified into one of four groups: HPV associated, HPV independent p53 wild type, HPV independent p53 abnormal, and indeterminate. Selected cases were further interrogated by (HPV RNA in situ hybridization,TP53sequencing).ResultsFinal subclassification yielded 22 (32.4%) HPV associated, 41 (60.3%) HPV independent p53 abnormal, and 5 (7.3%) HPV independent p53 wild type tumors. Interobserver agreement (overall Fleiss’ kappa statistic) for the four category classification was 0.74. No statistically significant differences in clinical outcomes between HPV associated and HPV independent vulvar squamous cell carcinoma were observed.ConclusionInterobserver reproducibility of vulvar squamous cell carcinoma subclassification based on p16 and p53 immunohistochemistry may support routine use in clinical practice. Vulvar field resection surgery showed no significant difference in clinical outcomes when stratified based on HPV status.

Rationale and Objectives The present study used diffusion-weighted imaging (DWI) to elucidate possible associations with tumor-infiltrating lymphocytes (TIL), tumor-stroma ratio and vimentin expression in head and neck squamous cell cancer (HNSCC). Materials and Methods 30 patients with primary HNSCC of different localizations were involved in the study. DWI was obtained on a 3 T scanner. Apparent diffusion coefficients (ADC) images were analyzed with a whole lesion measurement using a histogram approach. TIL- and vimentin-expression was calculated on biopsy samples before any form of treatment. Results Tumor-stroma ratio correlated with ADC kurtosis (r = 0.46, p = 0.01) and ADC skewness (r = 0.42, p = 0.02). Several ADC parameters were significantly different between stroma rich und tumor rich tumors. ADC entropy correlated significantly with the expression of TIL within the tumor compartment (r = 0.44, p = 0.01). No associations were identified between ADC parameters and vimentin expression. Conclusion ADC skewness and kurtosis histogram parameters can reflect tumor compartments in HNSCC. ADC entropy was associated with TIL of the tumor compartment but not with those of the stroma compartment, which emphasizes the ability of ADC histogram parameters to reflect distinctive differences of tumors.

Separate depiction of immunohistochemical staining of p16INK4a in situ.

List of transgenic mice used in the study.

Supplementary Figure legends

Reduction of β-catenin (CTNNB1) destroying complex components, for example, adenomatous polyposis coli (APC), induces β-catenin signaling and subsequently triggers activation of genes involved in proliferation and tumorigenesis. Though diminished expression of APC has organ-specific and threshold-dependent influence on the development of liver tumors in mice, the molecular basis is poorly understood. Therefore, a detailed investigation was conducted to determine the underlying mechanism in the development of liver tumors under reduced APC levels. Mouse liver at different developmental stages was analyzed in terms of β-catenin target genes including Cyp2e1, Glul, and Ihh using real-time RT-PCR, reporter gene assays, and immunohistologic methods with consideration of liver zonation. Data from human livers with mutations in APC derived from patients with familial adenomatous polyposis (FAP) were also included. Hepatocyte senescence was investigated by determining p16INK4a expression level, presence of senescence-associated β-galactosidase activity, and assessing ploidy. A β-catenin activation of hepatocytes does not always result in β-catenin positive but unexpectedly also in mixed and β-catenin–negative tumors. In summary, a senescence-inducing program was found in hepatocytes with increased β-catenin levels and a positive selection of hepatocytes lacking p16INK4a, by epigenetic silencing, drives the development of liver tumors in mice with reduced APC expression (Apc580S mice). The lack of p16INK4a was also detected in liver tumors of mice with triggers other than APC reduction.Implications: Epigenetic silencing of p16Ink4a in selected liver cells bypassing senescence is a general principle for development of liver tumors with β-catenin involvement in mice independent of the initial stimulus. Mol Cancer Res; 13(2); 239–49. ©2014 AACR.

Background: Patients with hormone-receptor-positive (HR+) breast cancer are at increased risk for late recurrence. One reason might be disseminated tumor cells (DTCs), which split off in the early stages of the disease and metastasize into the bone marrow (BM). Methods: We developed a novel multi-parameter immunofluorescence staining protocol using releasable and bleachable antibody–fluorochrome-conjugates. This sequential procedure enabled us to analyze six distinct phenotypical and therapy-related markers on the same DTC. We characterized BM aspirates from 29 patients with a HR+ tumor and a known positive DTC status—based on the standardized detection of epithelial cells in BM. Results: Using the immunofluorescence staining, a total of 153 DTCs were detected. Luminal A patients revealed a higher DTC count compared with luminal B. The majority of the detected DTCs were CK-positive (128/153). However, in 16 of 17 luminal A patients we found HER2-positive DTCs. We detected CK-negative DTCs (25/153) in 12 of 29 patients. Of those cells, 76% were Ki67-positive and 68% were HER2-positive. Moreover, we detected DTC clusters consisting of mixed characteristics in 6 of 29 patients. Conclusions: Using sequential multi-parameter imaging made it possible to identify distinct DTC profiles not solely based on epithelial features. Our findings indicate that characterization rather than quantification of DTCs might be relevant for treatment decisions.

ZusammenfassungDie vorliegende Übersicht fasst die relevanten Aspekte der S2k-Leitlinie Endometriose zusammen. Die Empfehlungen umfassen die Aufarbeitung und Befunderhebung bei Biopsien und Resektaten, die bei der klinischen Diagnose einer Endometriose entnommen wurden. Die Leitlinie berücksichtigt neben praktischen Aspekten der Pathologie ebenso die klinischen Notwendigkeiten an die Histopathologie für eine optimale Diagnostik und Therapie der Patientinnen.Basierend auf der in der pathologischen Literatur gebräuchlichsten Definition der Endometriose des Corpus uteri (Adenomyosis uteri) wurde diese in der Leitlinie definiert als der Nachweis des Endometrioseherdes im Myometrium in einem Abstand zur endomyometranen Grenze von einem mittelgroßen Gesichtsfeld (100fache Vergrößerung), was metrisch rund 2,5 mm entspricht. Bei Darmresektaten soll zum Status der Resektionsränder Stellung genommen werden.Ebenso definiert werden Anforderungen im Kontext endometrioseassoziierter Karzinome (z. B. Hormonrezeptorbestimmungen, immunhistochemische Untersuchung auf Ausfall der DNA-Mismatch-Reparaturproteine).

The 2020 WHO classification is focused on the distinction between HPV-associated and HPV-independent squamous cell carcinoma of the lower female genital organs. Differentiating according to HPV association does not replace the process of grading; however, the WHO classification does not recommend any specific grading system. VIN are also differentiated according to whether they are HPV(p16)-associated. HPV-independent adenocarcinoma (AC) of the cervix uteri has an unfavorable prognosis. Immunohistochemical p16 expression is considered to be a surrogate marker for HPV association. HPV-associated AC of the cervix uteri is determined using the prognostically relevant Silva pattern.

Purpose Epithelial-mesenchymal transition (EMT) is associated with increased metastatic spread and poor prognosis. Data on vulvar carcinoma are limited. Methods Thirty-two cases of squamous cell carcinoma of the vulva (16 with and 16 without inguinal lymph node metastases) and their lymph node deposits were evaluated for immunohistochemical expression of EMT markers (vimentin, cyclin D1, e-cadherin), p16, p53 and Ki-67. Results of EMT-immunostainings were compared to lymph node involvement and expression of p53 and p16. The micro-anatomical staining pattern for EMT markers comparing the tumor center with the front of invasion was analysed in each tumor. Results There was no difference in the expression of EMT markers between node negative and node positive tumors. Staining for vimentin and cyclin D1 was seen within tumor cells at the front of invasion in 100 and 84.4% of the tumors, respectively. The majority of cases (68.7%) showed negative or reduced staining for e-cadherin in this micro-anatomical localization. Tumor cells within the lymph node metastases showed positive staining for e-cadherin in 75% and for cyclin D1 in 49% of the cells but were negative for vimentin in 13 out of 16 cases (81.3%). Tumors with aberrant p53 staining represented a non-significant higher vimentin but significantly higher cyclin D1 expression at the front of invasion than those with p53 wild-type pattern. Conclusion The present study shows no differences in the expression of EMT markers between node positive and node negative vulvar cancers. The evaluation of immunostaining within the micro-anatomical context indicates that an EMT-phenotype is restricted to the tumor cells at the front of invasion. Paired analyses of vulvar carcinomas and their lymph node deposits suggest mesenchymal-epithelial transition (MET) in the metastatic deposits. Immunohistochemical staining results may suggest that EMT is more prevalent in vulvar cancer with aberrant p53 staining.

Purpose Histogram analysis can better reflect tumor heterogeneity than conventional imaging analysis. The present study analyzed possible correlations between histogram analysis parameters derived from Intravoxel-incoherent imaging (IVIM) and histopathological features in rectal cancer (RC). Methods Seventeen patients with histopathologically proven rectal adenocarcinomas were retrospectively acquired. In all cases, pelvic MRI was performed. Diffusion weighted imaging was obtained using a multi-slice single-shot echo-planar imaging sequence with b values of 0, 50, 200, 500 and 1000 s/mm2. Simplified IVIM analysis was performed using the IntelliSpace portal, version 10 and the following images were generated: f (perfusion fraction) map, D (true diffusion coefficient) map, and ADC map utilizing all b-values. Histogram based analysis of signal intensities was performed for every IVIM map using an in-house matlab tool. Histopathology was investigated using Ki 67 specimens with calculation of Ki 67-index and cellularity. CD31 stained specimens were used for calculation of microvessel density (MVD). Results There were statistically significant correlations between Ki 67 index and mode derived from ADC as well as entropy from f, r=−0.50, p=.04 and r=−0.55, p=.02, respectively. MVD correlated well with parameters derived from f. Conclusion IVIM histogram analysis parameters can reflect histopathology in RC. ADC and D values are associated with proliferation potential. Perfusion fraction f is associated with MVD.

Background Immunotherapy has become a cornerstone of the modern cancer treatment. It might be crucial to predict its expression non-invasively by imaging. The present study used diffusion-weighted imaging (DWI) quantified by whole lesion apparent diffusion coefficient (ADC) values to elucidate possible associations with programmed cell death ligand 1(PD-L1) expression in head and neck squamous cell cancer (HNSCC). Patients and methods Overall, 29 patients with primary HNSCC of different localizations were involved in the study. DWI was obtained by using a sequence with b - values of 0 and 800 s/mm2 on a 3 T MRI. ADC values were evaluated with a whole lesion measurement and a histogram approach. PD-L1 expression was estimated on bioptic samples before any form of treatment using 3 scores, tumor positive score (TPS), immune cell score (ICS), and combined positive score (CPS). Results An inverse correlation between skewness derived from ADC values and ICS was identified (r = -0.38, p = 0.04). ADCmax tended to correlate with ICS (r = -0.35, p = 0.06). Other ADC parameters did not show any association with the calculated scores. Conclusions There is a weak association between skewness derived from ADC values and PD-L1 expression in HNSCC, which might not be strong enough to predict PD-L1 expression in clinical routine. Presumably, ADC values are more influenced by complex histopathology compartments, comprising cellular and extracellular aspects of tumors than only of a single subset of tumor associated cells.

Primary vaginal carcinoma is rare. There are two pathogenetic pathways, one associated with HPV high-risk infection and another one with inactivation of p53. Vaginal Paget's disease is rare and mostly associated with vulvar disease or represents intravaginal spread of associated locoregional cancer. Diagnostic vaginal biopsies should be examined by step sections on H&E. Sentinel lymph nodes should be processed completely using ultrastaging. Morphology-based prognostic factors with good clinical evidence are tumour stage and lymph node status. Molecular markers are not currently relevant for treatment decision and prognosis.

Accurate disease classification is fundamental for the selection of the treatment approach, prognostication, selection of clinical trials and for research purposes in routine clinical practice. Extrauterine high-grade serous carcinoma (HG-SC) may arise from the ovary, the fallopian tube and rarely from the peritoneal surface epithelium. Regardless of its origin, the vast majority of patients with HG-SC share clinical symptoms, present with advanced stage disease and suffer from a poor prognosis. Recent data suggest that there is an increasing incidence of HG-SC arising from the fallopian tube. Data from the Clinical Cancer Registry of Leipzig of surgically treated non-uterine pelvic carcinomas were analyzed regarding their sites of origin. Depending on the histology, cases were separated into high-grade serous and non-high-grade serous tumors. Based on different approaches in the assessment of the site of origin, three distinct time periods were defined. The frequency of the specific sites of origin was compared to the different time periods and histologic subtypes. The majority of cases (57.9%; 279/482) were high-grade serous carcinomas, 42.1% of the cases presented with endometrioid, clear cell or mucinous histology. Overall, a 1.7-fold decrease of carcinomas with ovarian origin, paralleled by a 10.3-fold increase of tubal carcinomas was noted between 2000 and 2019. Based on the histopathological subtype, there was a 2.1-fold decrease of ovarian and a 7.1-fold increase of tubal carcinomas in patients with HG-SC. In non-high-grade serous tumors, the frequency of the different sites of origin did not change. 83.7% of tumors with non-high-grade serous histology originated from the ovary, whereas 86.8% of the carcinomas with tubal origin were of high-grade serous histology. The present and published data of non-uterine pelvic cancers may suggest an increase of tubal and decrease of ovarian carcinomas. However, there is rising morphologic and molecular evidence that non-uterine HG-SC actually arise from the fallopian tubes via its precursor STIC instead of from the ovary. This evidence has had an impact on the handling and reporting of non-uterine surgical specimens and its definition of the site assessment. In conclusion, the increasing frequency of tubal carcinomas and the associated decrease in ovarian cancer appears to be due to the reclassification of tumors previously classified as ovarian and greater emphasis on examining the resection specimens of non-uterine pelvic carcinomas.

Uterine sarcomas are a heterogeneous group of rare malignancies. Mostly (40-50%), they are leiomyosarcomas, followed by endometrial stromal sarcomas (ESS), low-grade (LG) and high-grade (HG), as well as undifferentiated sarcoma of the uterus (UUS) and adenosarcomas (AS). Other, non-organ-specific tumours such as NTRK-rearranged spindle cell neoplasia, perivascular epithelioid cell tumour (PEComa) and inflammatory myofibroblastic tumour (IMT) are extremely difficult to differentiate.In the most recent WHO classification, endometrial stromal tumours are subdivided as follows: benign, expansively growing endometrial stromal nodule (ESN) with sharp demarcation, the histologically similar-looking LG-ESS with infiltrative growth, the highly malignant HG-ESS and, as a diagnosis of exclusion, the highly aggressive UUS lacking specific lines of differentiation. LG-ESS can be differentiated from HG-ESS in most cases histomorphologically and immunohistochemically, but molecular investigations are necessary in individual cases. HG-ESS can be divided into 4 subtypes (YWHAE/NUTM2 fusion low-grade component, YWHAE/NUTM2 fusion high-grade component, ZC3H7B-BCOR fusion or BCOR-ITD) on the basis of molecular findings. Prognostically unfavourable factors in AS are severe sarcomatous overgrowth, deep myometrial invasion, high-grade histology and lymphatic vessel invasion. Tumours with NTRK fusion are immunohistochemically positive for S100 and TRK. PEComas express cathepsin K and HMB45, as well as TFE3 when translocation is present. Almost every IMT shows an alteration in the ALK gene In the case of overlapping morphology and simultaneous therapeutic and prognostic relevance, it is becoming increasingly important to verify or confirm the suspected histomorphological diagnosis by immunohistochemical and possibly molecular investigations.Uterine Sarkome sind eine heterogene Gruppe seltener Malignome. Meist (40–50 %) handelt es sich um Leiomyosarkome, gefolgt von endometrialen Stromasarkomen (ESS), low-grade (LG) und high-grade (HG), sowie undifferenzierten Sarkomen des Uterus (UUS) und Adenosarkomen (AS). Differenzialdiagnostisch äußerst schwierig sind einzelne, nicht organspezifische Tumoren wie die NTRK-rearrangierte Spindelzellneoplasie, der primäre perivaskuläre epitheloidzellige Tumor (PECom) und der inflammatorische myofibroblastäre Tumor (IMT).In der jüngsten WHO-Klassifikation werden endometriale Stromatumoren unterteilt in: benigner, expansiv wachsender endometrialer Stromaknoten (ESN) mit scharfer Begrenzung, das histologisch gleichartig aussehende LG-ESS mit infiltrativem Wachstum und das hochmaligne HG-ESS sowie als Ausschlussdiagnose das hochaggressive UUS ohne Liniendifferenzierung. Das LG-ESS lässt sich meist histologisch und immunhistochemisch vom HG-ESS unterscheiden, molekulare Untersuchungen sind in Einzelfällen jedoch notwendig. Das HG-ESS kann auf Basis molekularer Befunde in 4 Subtypen (YWHAE/NUTM2-Fusion low-grade Komponente, YWHAE/NUTM2-Fusion high-grade Komponente, ZC3H7B-BCOR-Fusion oder BCOR-ITD) unterteilt werden. Prognostisch ungünstige Faktoren beim AS sind: starke sarkomatöse Überwucherung, tiefe Myometriuminvasion, high-grade Histologie, Lymphgefäßinvasion. Tumoren mit NTRK-Fusion sind immunhistochemisch positiv für S100 und TRK. Das PECom exprimiert Cathepsin K und HMB45 sowie TFE3 bei vorhandener Translokation. Nahezu jedes IMT weist eine Alteration im ALK-Gen auf. Eine Absicherung durch immunhistochemische und eventuell molekulare Untersuchungen ist bei überschneidender Morphologie und gleichzeitiger therapeutisch und prognostischer Relevanz von immer größerer Bedeutung.

The 2020 WHO Classification defines the spindle cell, epithelioid, and myxoid variants as subtypes of uterine leiomyosarcomas (LMS). Presence of cellular atypia (size variation of polymorphic nuclei 2-3:1), tumor cell necroses, and mitotic count (usually ≥ 10 MF/10 HPF) are still the key features for diagnostic separation from uterine leiomyomas. Preanalytic variables, staining quality, as well as intralesional geographic distribution may affect the mitotic count. Smooth muscle tumors of uncertain malignant potential (STUMP) still exist as a not yet well-characterized diagnostic entity. Immunohistochemical stains against p16, p53, Ki-67, and WT‑1 may aid differential diagnosis in selected cases. Diagnostic molecular pathology is not yet relevant for diagnosis.Die WHO-Klassifikation 2020 unterscheidet das spindelzellige, epitheloide und myxoide Leiomyosarkom (LMS) als morphologische Varianten. Diagnostische Parameter zur Unterscheidung zum Leiomyom sind nach wie vor nukleäre Atypien (Kerngrößenschwankungen 2–3: 1), die Mitosezahl (meist ≥ 10 Mitosen/10 HPF bei 0,55 mm Gesichtsfelddurchmesser bzw. 0,24 mm

Congenital anomalies of the kidney and urinary tract (CAKUT) represent the most common cause of chronic kidney failure in children. Despite growing knowledge of the genetic causes of CAKUT, the majority of cases remain etiologically unsolved. Genetic alterations in roundabout guidance receptor 1 (ROBO1) have been associated with neuronal and cardiac developmental defects in living individuals. Although Slit-Robo signaling is pivotal for kidney development, diagnostic ROBO1 variants have not been reported in viable CAKUT to date. By next-generation-sequencing methods, we identified six unrelated individuals and two non-viable fetuses with biallelic truncating or combined missense and truncating variants in ROBO1. Kidney and genitourinary manifestation included unilateral or bilateral kidney agenesis, vesicoureteral junction obstruction, vesicoureteral reflux, posterior urethral valve, genital malformation, and increased kidney echogenicity. Further clinical characteristics were remarkably heterogeneous, including neurodevelopmental defects, intellectual impairment, cerebral malformations, eye anomalies, and cardiac defects. By in silico analysis, we determined the functional significance of identified missense variants and observed absence of kidney ROBO1 expression in both human and murine mutant tissues. While its expression in multiple tissues may explain heterogeneous organ involvement, variability of the kidney disease suggests gene dosage effects due to a combination of null alleles with mild hypomorphic alleles. Thus, comprehensive genetic analysis in CAKUT should include ROBO1 as a new cause of recessively inherited disease. Hence, in patients with already established ROBO1-associated cardiac or neuronal disorders, screening for kidney involvement is indicated.

Texture analysis derived from computed tomography (CT) might be able to provide clinically relevant imaging biomarkers and might be associated with histopathological features in tumors. The present study sought to elucidate the possible associations between texture features derived from CT images with proliferation index Ki-67 and grading in pulmonary neuroendocrine tumors. Overall, 38 patients (n = 22 females, 58%) with a mean age of 60.8 ± 15.2 years were included into this retrospective study. The texture analysis was performed using the free available Mazda software. All tumors were histopathologically confirmed. In discrimination analysis, “S(1,1)SumEntrp” was significantly different between typical and atypical carcinoids (mean 1.74 ± 0.11 versus 1.79 ± 0.14, p = 0.007). The correlation analysis revealed a moderate positive association between Ki-67 index with the first order parameter kurtosis (r = 0.66, p = 0.001). Several other texture features were associated with the Ki-67 index, the highest correlation coefficient showed “S(4,4)InvDfMom” (r = 0.59, p = 0.004). Several texture features derived from CT were associated with the proliferation index Ki-67 and might therefore be a valuable novel biomarker in pulmonary neuroendocrine tumors. “Sumentrp” might be a promising parameter to aid in the discrimination between typical and atypical carcinoids.

In 2021, the 2015 German consensus guideline for the diagnosis and treatment of uterine cervical carcinoma was updated. The present article summarises the new recommendations for pathologists: the incorporation of the International Endocervical Adenocarcinoma Classification (IECC), which morphologically separates HPV-associated and non-HPV-associated adenocarcinomas, as well as the reporting of the prognostic relevant growth pattern of the adenocarcinoma of the endocervical subtype (Silva pattern). Histologically, multifocality has been defined as the presence of clearly invasive foci with a minimum distance between each focus of 0.2 cm. Because of its intratumoural heterogeneity, all carcinomas ≤ 2 cm in their largest dimension should be processed completely, and tumours 2 cm should be processed with one block per centimetre of their greatest dimension. In cases of (radical) trachelectomy/hysterectomy, the distal vaginal resection margin and all parametrial tissue should be processed completely. Sentinel lymph nodes have to be processed completely by lamellation along its long axis in 0.2 cm intervals. Immunohistochemical ultrastaging is mandatory. Staging should be performed using the 2009 FIGO classification and 2017 TNM classification. Reporting the revised 2018 FIGO classification is optional. To date, molecular markers have not been relevant for prognostication and treatment decision making.Im Jahr 2015 wurde die S3-Leitlinie für Diagnostik und Therapie des Zervixkarzinoms erstellt, die 2021 in ihrer revidierten Fassung vorliegt. Neu für die Pathologie sind die Aufnahme der International Endocervical Adenocarcinoma Classification (IECC), die zwischen HPV-assoziierten und non-HPV-assoziierten Karzinomen unterschiedet, sowie des prognostisch relevanten morphologischen Wachstumsmusters bei (HPV-assoziierten) Adenokarzinomen (sog. Silva-Pattern). Die Multifokalität beim mikroinvasiven Zervixkarzinom ist nun definiert als der Nachweis voneinander histologisch klar separierter invasiver Foci mit einem minimalen Abstand von 0,2 cm. Aufgrund einer möglichen intratumoralen Heterogenität sollen Zervixkarzinome ≤ 2 cm vollständig und Tumoren 2 cm mit einem Block pro Zentimeter größter Ausdehnung eingebettet werden. Bei der (radikalen) Trachelektomie/Hysterektomie sollen der vaginale Resektionsrand und das parametrane Gewebe vollständig eingebettet werden. Neu ist die Empfehlung zur Aufarbeitung von Sentinellymphknoten. Diese sollen entlang ihrer Längsachse in 0,2 cm Abstand lamelliert, vollständig eingebettet und mittels Ultrastaging aufgearbeitet werden. Die FIGO-Klassifikation von 2009 bzw. TNM-Kategorisierung von 2017 wird beibehalten, die Angabe der FIGO-Klassifikation 2018/19 ist optional. Molekulare Marker spielen derzeit noch keine relevante Rolle in der Prognose und zum Therapieentscheid.

Introduction/Background* Vulvar squamous cell carcinoma (VSCC) is subclassified into three prognostically relevant groups: i) HPV-Associated (HPV-A), ii) HPV-Independent TP53 mutated (HPV-I p53abn), and iii) HPV-Independent TP53 wild type (HPV-I p53wt). Immunohistochemistry (IHC) for p16 and p53 are used as surrogate markers for HPV viral integration and TP53 mutational status, respectively. We assessed the reproducibility of subclassification based on p16 and p53 IHC and evaluated the prognostic significance of VSCC molecular subgroups in a patient cohort treated by vulvar field resection (VFR) surgery. Methodology VSCC formalin-fixed paraffin-embedded tumour tissue and patient clinicopathologic data from 68 cases treated by vulvar field resection (VFR) were collected from the Leipzig School of Radical Pelvic Surgery. Immunohistochemistry for p16 and p53 was performed at three different institutions and evaluated independently by seven pathologists and two pathology research fellows. Tumors were classified into one of four groups: HPV-A, HPV-I p53wt, HPV-I p53abn, and Indeterminate (requiring additional investigation). Selected cases were further interrogated by HPV RNA in situ hybridisation (ISH) and TP53 sequencing. Result(s)* Final VSCC subclassification yielded 22 tumors (32.4%) were HPV-A, 41 (60.3%) were HPV-I p53abn, and 5 (7.3%) were HPV-I p53wt; the clinicopathologic data is summarised in table 1. Interobserver agreement (overall Fleiss’ kappa statistic) for the four-category classification was 0.74. No statistically significant differences in clinical outcomes between HPV-A and HPV-I VSCC were observed (figure 1). Conclusion* Interobserver reproducibility of VSCC subclassification based on p16 and p53 IHC is very good, supporting their routine use in clinical practice. VSCCs treated by VFR surgery showed no significant difference in clinical outcomes when stratified based on HPV status. This contrasts with the significant prognostic differences between HPV-A and HPV-I VSCC demonstrated in cohorts treated by conventional conservative surgical approaches and suggests that a change in surgical approach can improve outcomes in patients with HPV-I VSCC.

Endometrial mesonephric-like adenocarcinoma (ML-AC) represents a recently recognized subtype of endometrial adenocarcinoma (AC) associated with a subtle immunophenotype with a characteristic KRAS-mutation. Detailed clinico-pathologic analyses and prognostic data on ML-AC are limited. We report a series of four cases with histopathological, immunohistochemical, and molecular analyses. These cases as well as the data of previously published cases were reviewed for clinico-pathologic variables and clinical follow-up information. Forty cases of ML-AC were identified. ML-AC represents about 1% of all endometrial carcinomas. Similar to other types of endometrial AC, vaginal bleeding was the leading presenting symptom, and the mean age was 60.0 years (range 31–91). More than a half of the patients presented with locally advanced disease (≥ FIGO stage II) at time of diagnosis, developed a recurrence or died of the disease within a mean follow-up period of 24.7 months (range 3–144.5 months). The most common site of distant disease was pulmonary involvement. Microscopically, ML-ACs present with mixed morphology and show a co-expression of so-called mesonephric and Mullerian markers, suggesting a Mullerian origin of the tumors. Immunostaining for PD-L1 was negative in all tested cases, using different antibodies against PD-L1. Retained staining for mismatch repair proteins on immunohistochemistry and a POLE-mutation suggest a copy number low phenotype within the molecular classification of endometrial carcinomas. Almost all cases showed a KRAS-mutation at codon 12 (mostly G12V). Uterine ML-AC represents a distinct subtype of invasive endometrial AC, associated with KRAS-mutations and characteristic immunohistochemical findings. Clinically, ML-AC may show an aggressive behavior with a high rate of recurrent disease and a substantial risk for distant metastatic disease, especially to the lungs.

Zahlreiche therapie- und prognoseorientierte Studien zum Zervixkarzinom haben eine Uberarbeitung der FIGO-Klassifikation erforderlich gemacht. Das mikroinvasive Karzinom wird allein durch die Tiefe der Stromainfiltration definiert, die flachige (horizontale) Ausdehnung ist nunmehr irrelevant. Makroinvasive Karzinome, die auf die Cervix uteri begrenzt sind, werden anhand der Tumorgrose subkategorisiert in: FIGO IB1/T1b1 – zervikale Stromainvasion >5,0 mm und ≤2 cm maximale Tumorgrose, FIGO IB2/T1b2 – Tumorgrose >2 cm und ≤4 cm, FIGO IB3/T1b3 – Tumorgrose >4 cm. Sowohl die pelvinen als auch die paraaortalen Lymphknoten werden als regionare Lymphknoten definiert. Der Befall pelviner Lymphknoten wird als FIGO IIIC1/pN1a und eine Metastasierung in paraaortale Lymphknoten mit und ohne Beteiligung pelviner Lymphknoten als FIGO IIIC2/pN1b klassifiziert. Der Nachweis einer Corpusinfiltration und eine adnexale/ovarielle Metastasierung sind nicht stagingrelevant.

Background and objectives Texture analysis derived from morphological magnetic resonance (MR) images might be associated with histopathology in tumors. The present study sought to elucidate possible associations between texture features derived from T1-and T2-weighted images with proliferation index Ki67 in soft tissue sarcomas. Methods Overall, 29 patients (n = 13, 44.8% female) with a median age of 52 years were included into this retrospective study. Several soft tissue sarcomas were investigated. Texture analysis was performed on pre-contrast T1-weighted and T2-weighted images using the free available Mazda software. Results The best correlation coefficients with Ki67 index were identified for the following parameters: T1-weighted images “45dgr_RLNonUni (p = 0.50, P = 0.006), T2-weighted images “S (4,0)SumAverg” (p = −0.45, P = 0.02). A ROC analysis was performed for Ki67-index with a threshold of 10%. The highest area under the curve (AUC) was found for the parameter “T1_WavEnHL_s-7” with an AUC of 0.90. For the threshold of Ki67 = 20% the highest AUC was identified for the parameter „T2_S (1,1)Entropy” with an AUC of 0.77. Conclusion Several texture features derived from T1-and T2-weighted images correlated with proliferation index Ki67 and might be used as valuable novel biomarkers in soft tissue sarcomas.

Objective Diffusion weighted imaging (DWI) can be quantified by apparent diffusion coefficient (ADC) and can predict tissue microstructure. The aim of the present study was to analyze possible associations between ADC histogram based parameters with different histopathological parameters in cervical squamous cell carcinoma. Materials and methods 18 female patients (age range 32–79 years) with squamous cell cervical carcinoma were retrospectively enrolled. In all cases, pelvic MRI was performed with a DWI (b-values 0 and 1000 s/mm2). Histogram analysis was performed as a whole lesion measurement. Histopathological parameters included expression of EGFR, VEGF, Hif1-alpha, Her2 and Histone 3. Spearman's correlation coefficient was used to analyze associations between investigated parameters. Results Analyze of the investigated ADC histogram parameters showed a good interreader variability, ranging from 0.705 for entropy to 0.959 for ADCmedian. EGFR expression correlated statistically significant with several histogram parameters. The highest correlation was observed for p75 (p = −0.562, P = 0.015). There were several correlations with histone 3, the highest with p25 (p = −0.610, P = 0.007). None of the ADC related parameters correlated statistically significant with expression of VEGF, Hif1-alpha and Her2. Conclusion Histogram analysis showed a good interreader agreement. ADC histogram parameters might be able to reflect expression of EGFR and histone 3 in cervical squamous cell carcinomas, but not expression of VEGF, Hif1-alpha and Her2.

Background Pregnant women are also susceptible to SARS-CoV-2. Although an infection of the placenta may be rare, pregnancy may occasionally be affected by intrauterine failure. The knowledge of placental morphology on sudden intrauterine demise is still limited. Methods Fetal and placental tissue of two cases of sudden intrauterine death in the second trimester were analysed morphologically and by immunohistochemistry. One case was evaluated by RT-PCR. Results Both mothers were tested positive for the Alpha variant of SARS-CoV-2 but were oligosymptomatic for COVID-19. Unexpected sudden intrauterine death (SIUD) occurred at 15 + 2 and 27 + 3 weeks of gestation. One fetus demonstrated an intrauterine growth restriction. No malformations nor inflammatory changes were observed in either fetus on autopsy. In contrast to the placentas, the fetal tissue was negative for SARS-CoV-2 on immunohistochemical and RT-PCR analyses. Macroscopically, the placentas showed an increased consistency with a white, reticular cutting surface covering about 95% of the whole placenta. Only very focal histiocytic chronic intervillositis was noted histologically. Massive perivillous fibrin deposits with extensive necroses of the villous trophoblast were present in more than 90% of the placental tissue. Immunohistochemical staining was strong and diffusely positive for SARS-CoV-2 in the villous trophoblast and rarely within the villous stromal cells. Placental SARS-CoV-2 infection was confirmed by RT-PCR. Conclusion Sudden intrauterine death may occur in mothers who are oligosymptomatic for COVID-19. Acute placental failure is responsible for SIUD, demonstrated by massive perivillous fibrin deposits and extensive necroses of the villous trophoblast with SARS-CoV-2-positivity based on immunohistochemical staining and RT-PCR. Detailed histopathological examination of placental and fetal tissue is mandatory to verify SARS-CoV-2 and to evaluate the pathogenesis and functionality of this disease.

The present article summarises the recommendations for the handling, histopathological workup, diagnostics and reporting in surgical pathology of biopsies and resection specimens in patients with the clinical diagnosis of endometriosis. In addition to practical aspects of pathology, the guidelines also take into account the clinical requirements for histopathology for the optimal diagnosis and therapy of the patients.Based on the definition of endometriosis of the corpus uteri (adenomyosis uteri) most commonly used in the pathological literature, this was defined in the guidelines as the detection of the endometriosis focus in the myometrium at a distance from the endomyometrial border of a medium-sized visual field (100× magnification), which in metric units corresponds to around 2.5 mm. In bowel resection specimens, the status of the resection margins had to be documented within the histopathological report.Also mentioned are the requirements for the reporting of carcinomas associated with endometriosis, including the immunohistochemical evaluation of steroid hormone receptors and mismatch repair proteins.Die vorliegende Übersicht fasst die relevanten Aspekte der S2k-Leitlinie Endometriose zusammen. Die Empfehlungen umfassen die Aufarbeitung und Befunderhebung bei Biopsien und Resektaten, die bei der klinischen Diagnose einer Endometriose entnommen wurden. Die Leitlinie berücksichtigt neben praktischen Aspekten der Pathologie ebenso die klinischen Notwendigkeiten an die Histopathologie für eine optimale Diagnostik und Therapie der Patientinnen.Basierend auf der in der pathologischen Literatur gebräuchlichsten Definition der Endometriose des Corpus uteri (Adenomyosis uteri) wurde diese in der Leitlinie definiert als der Nachweis des Endometrioseherdes im Myometrium in einem Abstand zur endomyometranen Grenze von einem mittelgroßen Gesichtsfeld (100fache Vergrößerung), was metrisch rund 2,5 mm entspricht. Bei Darmresektaten soll zum Status der Resektionsränder Stellung genommen werden.Ebenso definiert werden Anforderungen im Kontext endometrioseassoziierter Karzinome (z. B. Hormonrezeptorbestimmungen, immunhistochemische Untersuchung auf Ausfall der DNA-Mismatch-Reparaturproteine).

Purpose The present study used histogram analysis values derived from T1- and T2- weighted (w) images to elucidate possible associations with Tumor-infiltrating lymphocytes (TIL) and Vimentin expression in head and neck squamous cell cancer (HNSCC). Materials and methods Overall, 28 patients (n = 8 female patient, 28.6%) with primary HNSCC of different localizations were involved in the study. Magnetic resonance imaging (MRI) was obtained on a 3 T MRI. The images were analyzed with a whole lesion measurement using a histogram approach. TIL- and vimentin-expression was calculated on biopsy samples before any form of treatment. Results Several T1-derived parameters correlated with the expression of TIL within the stroma compartment: mean (r = 0.42, p = 0.025), p10 (r = 0.50, p = 0.007), p25 (r = 0.42, p = 0.025), median (r = 0.39, p = 0.036), and mode (r = 0.39, p = 0.04). No T2-derived parameter correlated with the TIL within the stroma compartment. Several T2-derived parameters correlated with the expression of TIL within the tumor compartment: mean (r = −0.52, p = 0.004), max (r = −0.43, p = 0.02), p10 (r = −0.38, p = 0.04), p25 (r = −0.53, p = 0.004), p75 (r = −0.52, p = 0.004), p90 (r = −0.48, p = 0.009), median (r = −0.52, p = 0.004), mode (r = −0.40, p = 0.03). Kurtosis derived from T2w images had significant higher values in tumor-rich tumors, compared to stroma-rich tumors, (mean 5.5 ± 0.5 versus 4.2 ± 1.2, p = 0.028). Conclusions Histogram analysis parameters derived from T1w and T2w images might be able to reflect tumor compartments and TIL expression in HNSCC.

The new WHO classification of tumors of the female genitalia entails some changes, especially those of prognostic and therapeutic relevance: there is a return to the term borderline tumor. Implants are again subdivided into noninvasive implants of the epithelial or desmoplastic type as before. Invasive extraovarian implants are classified as low-grade serous carcinoma (LGSC). Former seromucinous carcinomas are now classified as endometrioid carcinomas (seromucinous subtype). New entities of ovarian carcinomas are mesonephric-like adenocarcinoma, undifferentiated and dedifferentiated carcinoma, and mixed carcinoma. The classification of neuroendocrine neoplasms is analogous to that of pulmonary and gastrointestinal neuroendocrine neoplasms, regardless of their location. Endometrioid endometrial carcinoma can be classified into four molecular subtypes, which have significant prognostic significance. New subtypes include mucinous carcinoma of the intestinal type and mesonephric-like adenocarcinoma. Stromasarcomas of the endometrium are further subclassified based on specific molecular alterations. Adenocarcinomas (ACs) and squamous cell carcinomas (PECs) of the lower female genital tract are distinguished from HPV-associated and HPV-independent carcinomas. Block-like staining for p16 is the accepted surrogate immunohistochemical marker. Grading has not been reported for PEC. For HPV-associated AC of the cervix uteri, prognostic assessment is based on the pattern of invasion (so-called Silva pattern). Serous carcinomas in the cervix uteri are endometrial carcinomas with cervical infiltration.Die neue WHO-Klassifikation der Tumoren des weiblichen Genitale bringt einige, insbesondere prognostisch und therapeutisch relevante Änderungen mit sich: Es erfolgt eine Rückkehr zum Begriff Borderlinetumor. Implantate werden wieder wie früher unterteilt in nichtinvasive Implantate vom epithelialen oder vom desmoplastischen Typ. Invasive extraovarielle Implantate werden als low-grade seröses Karzinom (LGSC) klassifiziert. Die ehemaligen seromuzinösen Karzinome gehören jetzt zu den endometrioiden Karzinomen (seromuzinöser Subtyp). Neue Entitäten der Ovarialkarzinome sind das Mesonephric-like-Adenokarzinom, das undifferenzierte und dedifferenzierte Karzinom sowie das gemischtes Karzinom. Die Klassifikation der neuroendokrinen Neoplasien erfolgt unabhängig ihrer Lokalisation analog der pulmonalen und gastrointestinalen neuroendokrinen Neoplasien. Das endometrioide Endometriumkarzinom kann in 4 molekulare Subtypen klassifiziert werden, welchen eine wesentliche prognostische Bedeutung zukommt. Als neue Subtypen sind das muzinöse Karzinom vom intestinalen Typ und das Mesonephric-like-Adenokarzinom hinzugekommen. Stromasarkome des Endometriums werden anhand bestimmter molekularer Alterationen weiter subklassifiziert. Bei Adeno- (AC) und Plattenepithelkarzinomen (PEC) des unteren weiblichen Genitales werden HPV-assoziierte von HPV-unabhängigen Karzinomen unterschieden. Eine blockartige Färbung für p16 ist der akzeptierte immunhistochemische Surrogatmarker. Ein Grading ist für PEC nicht angegeben. Bei HPV-assoziierten AC der Cervix uteri erfolgt die prognostische Einschätzung mittels des Invasionsmusters (sog. Silva-Pattern). Bei serösen Karzinomen in der Cervix uteri handelt es sich um Endometriumkarzinome mit Zervixinfiltration.

Primary vaginal carcinoma is rare. There are two pathogenetic pathways, one associated with HPV high-risk infection and another one with inactivation of p53. Vaginal Paget's disease is rare and mostly associated with vulvar disease or represents intravaginal spread of associated locoregional cancer. Diagnostic vaginal biopsies should be examined by step sections on HE. Sentinel lymph nodes should be processed completely using ultrastaging. Morphology-based prognostic factors with good clinical evidence are tumour stage and lymph node status. Molecular markers are not currently relevant for treatment decision and prognosis.Das Vaginalkarzinom ist ein seltenes Malignom des weiblichen Genitales. Pathogenetisch können ein HPV-high-risk-Pathogeneseweg und ein p53-abhängiger Pathogeneseweg definiert werden. Der vaginale M. Paget ist selten und zumeist assoziiert mit einem M. Paget der Vulva oder lokoregionären Karzinomen. Diagnostische Biopsien sollen in Stufenschnitten aufgearbeitet werden. Sentinel-Lymphknoten sollen vollständig und mittels Ultrastaging untersucht werden. Etablierte morphologische Prognosefaktoren sind das Tumorstadium und lokoregionäre Metastasen. Molekulare Marker spielen derzeit keine Rolle.

Perifocal edema of brain tumors is associated with survival and neurological symptoms. Our aim was to analyze associations between perifocal edema and immunohistochemical features including proliferation potential, microvessel density, neoangiogenesis and invasiveness in brain metastasis (BM).35 patients with BM were included into the retrospective study. The tumors were localized supratentorial in 25 lesions (71.4%) and infratentorial in 10 lesions (28.6%). The following immunohistochemical features were calculated on histopathological specimens: microvessel density, proliferation index Ki 67, matrix-metallopeptidase 9 (MMP9) extracellular matrix metalloproteinase inducer (EMMPRIN) and vascular endothelial growth factor (VEGF) expression. Tumor and edema volumes were estimated semiautomatically on magnetic resonance images.There were no correlations between tumor volume and edema volume. Moreover, no correlation was identified between the investigated immunohistochemical features and tumor/edema volume. In the non-small cell lung cancer subgroup, a positive correlation between tumor volume and VEGF expression was observed (r = 0.52, P = 0.02) and edema volume correlated inversely with MMP9 expression (r = -0.53, P = 0.02).In BM, no linear associations exist between tumor volumes, edema volumes and immunohistochemical features reflecting proliferation potential, neoangiogenesis, microvessel density and MMP9 expression. However, in the subgroup of non-small cell lung cancer, there might be associations between MMP9 expression and edema volume as well as between tumor volume and angiogenesis.

Uterine sarcomas represent a heterogeneous group of rare malignancies, derived from the myometrium, the endometrial stroma, and very rarely from the nonspecialized uterine soft tissue. The actual incidence is about 1.5 for Caucasian and 3.0 for Afro-American women. There is no grading system for leimoysarcoma defined by the WHO classification; however, if clinicians request, the FNCLCC grading can be specified in analogy to soft tissue sarcomas. Adenosarcomas must be distinguished from adenofibromas (the existence of which is questionable)-with the vast majority of these tumors being uterine adenosarcomas. Within adenosarcomas, deep myometrial invasion (50%), sarcomatous overgrowth, and a high-grade heterologous component are associated with a higher recurrence rate and poor survival. The immunohistochemical panel represents a very helpful tool for distinguishing low-grade from high grade endometrial stromal sarcomas (ESS) and may be supplemented by molecular analyses. Steroid hormone receptor analysis should be performed for all ESS due to the possible therapeutic relevance. Undifferentiated uterine sarcomas represent a diagnosis of exclusion and have a very poor prognosis. Carcinosarcomas represent a special subtype of endometrial carcinomas and are in fact not uterine sarcomas. Uterine sarcomas may present substantial intratumoral heterogeneity and adequate embedding is mandatory. Lesions ≤2 cm in the largest dimension should be processed completely and larger tumors should be processed with one block per centimeter for the largest tumor dimension.