The presence of cancer stem cells is linked to aggressive disease and higher risk of recurrence, and multiple markers have been proposed to detect cancer stem cells. However, a detailed evaluation of the expression patterns and the prognostic value of markers relevant for endometrial cancer is lacking. As organoid models are suggested to be enriched in cancer stem cells, such models may prove valuable to define tissue-specific cancer stem cells. To address this, imaging mass cytometry and multiplex single-cell analyses were performed on an endometrial cancer patient series including both tumor biopsies and corresponding patient-derived organoids. An antibody panel focused on cancer stem cell markers was used to identify cancer stem cell phenotypes. Over 70% of epithelial cells in the tumor biopsies expressed at least one putative cancer stem cell marker. We identified distinct cancer cell phenotypes with heterogeneous expression within individual patients and between patient samples. Few differences in the distribution of cancer cell phenotypes were observed between tumor biopsies and corresponding organoids. Cells expressing aldehyde dehydrogenase 1 (ALDH1) were more prevalent in high-grade tumors, while expression of CD44 was more prevalent in grade 1 tumors. Spatial analysis revealed significantly less interaction between ALDH1- and CD44-expressing cells. Gene expression data was used to further investigate selected markers. CD44 gene expression was associated with a favorable prognosis and was further validated using immunohistochemistry. High expression of CD44 was significantly associated with better survival. The general high expression of proposed stem cell markers may indicate alternative roles for these in endometrial cancer.

Objective Although early-detected cervical cancer is associated with good survival, the prognosis for late-stage disease is poor and treatment options are sparse. Mismatch repair deficiency (MMR-D) has surfaced as a predictor of prognosis and response to immune checkpoint inhibitor(s) in several cancer types, but its value in cervical cancer remains unclear. This study aimed to define the prevalence of MMR-D in cervical cancer and assess the prognostic value of MMR protein expression. Methods Expression of the MMR proteins MLH-1, PMS-2, MSH-2, and MSH-6 was investigated by immunohistochemical staining in a prospectively collected cervical cancer cohort (n=508) with corresponding clinicopathological and follow-up data. Sections were scored as either loss or intact expression to define MMR-D, and by a staining index, based on staining intensity and area, evaluating the prognostic potential. RNA and whole exome sequencing data were available for 72 and 75 of the patients and were used for gene set enrichment and mutational analyses, respectively. Results Five (1%) tumors were MMR-deficient, three of which were of neuroendocrine histology. MMR status did not predict survival (HR 1.93, p=0.17). MSH-2 low (n=48) was associated with poor survival (HR 1.94, p=0.02), also when adjusting for tumor stage, tumor type, and patient age (HR 2.06, p=0.013). MSH-2 low tumors had higher tumor mutational burden (p=0.003) and higher frequency of (frameshift) mutations in the double-strand break repair gene RAD50 (p<0.01). Conclusion MMR-D is rare in cervical cancer, yet low MSH-2 expression is an independent predictor of poor survival.

Abstract Background Precise preoperative risk classification of endometrial cancer is crucial for treatment decisions. Existing clinical markers often fail to accurately predict lymph node metastasis and recurrence risk. Loss of vimentin expression has emerged as a potential marker for predicting recurrence in low-risk endometrial cancer patients. We assessed whether vimentin expression in preoperative biopsies predicts poor prognosis and lymph node metastasis in a large multicentre cohort. Methods Vimentin expression was evaluated using immunohistochemistry in 1483 patients diagnosed with endometrial cancer across 14 hospitals in Europe. Expression levels of vimentin were analyzed in conjunction with clinical characteristics for predicting disease-specific survival and lymph node metastases. Results Vimentin loss was significantly associated with aggressive disease and poor survival. Adjusted for clinicopathological variables, vimentin remained independently prognostic with a hazard ratio (HR) of 1.68 (95% CI 1.16–2.42, P = 0.006). Vimentin expression remained independently prognostic in endometrioid endometrial cancer- and FIGO staged 1 patient. Interestingly, vimentin loss independently predicted lymph node metastases, with an HR of 1.83 (95% CI 1.13–2.95, P = 0.014). Conclusions Loss of vimentin in preoperative biopsies serves as an independent predictor of poor prognosis and lymph node metastases. Incorporating vimentin as a clinical marker can improve risk stratification and treatment decisions.

The authors wish to point out an inadvertent error in Fig. 4 in the published article. The Kaplan–Meier curve given in Fig. 4i does not correspond to the clusters in 4 g/h. The correct Fig. 4i is presented below. The authors would like to emphasize that the error does not affect the description, interpretation or original conclusions. The authors sincerely apologize for any inconvenience caused by this error. Single-cell profiling of low-stage endometrial cancers identifies low epithelial vimentin expression as a marker of recurrent diseaseThis study reveals distinct characteristics in low-stage tumors and points to vimentin as a clinically relevant marker that may aid in identifying a here to unidentified subgroup of high-risk patients. Full-Text PDF Open Access

Background Identification of aggressive low-stage endometrial cancers is challenging. So far, studies have failed to pinpoint robust features or biomarkers associated with risk of recurrence for these patients.Methods Imaging mass cytometry was used to examine single-cell expression of 23 proteins in 36 primary FIGO IB endometrial cancers, of which 17 recurred. Single-cell information was extracted for each tumor and unsupervised clustering was used to identify cellular phenotypes. Distinct phenotypes and cellular neighborhoods were compared in relation to recurrence. Cellular differences were validated in a separate gene expression dataset and the TCGA EC dataset. Vimentin protein expression was evaluated by IHC in pre-operative samples from 518 patients to validate its robustness as a prognostic marker.Findings The abundance of epithelial, immune or stromal cell types did not associate with recurrence. Clustering of patients based on tumor single cell marker expression revealed distinct patient clusters associated with outcome. A cell population neighboring CD8+ T cells, defined by vimentin, ER, and PR expressing epithelial cells, was more prevalent in non-recurrent tumors. Importantly, lower epithelial vimentin expression and lower gene expression of VIM associated with worse recurrence-free survival. Loss and low expression of vimentin was validated by IHC as a robust marker for recurrence in FIGO I stage disease and predicted poor prognosis also when including all patients and in endometrioid patients only.Interpretation This study reveals distinct characteristics in low-stage tumors and points to vimentin as a clinically relevant marker that may aid in identifying a here to unidentified subgroup of high-risk patients.Copyright (c) 2023 The Author(s). Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

Introduction/Background Although early detected cervical cancer is associated with good survival, prognosis for late-stage disease is poor and treatment options are sparse. Mismatch-repair (MMR) deficiency has surfaced as a predictor of immune checkpoint inhibitor responses in several cancer types, but its value in cervical cancer remains unclear. This study aimed to define the incidence of MMR deficiency and establish its prognostic significance as well as the value of separate MMR proteins in cervical cancer. Methodology Expression of the MMR proteins MLH-1, PMS-2, MSH-2, MSH-6 was investigated by immunohistochemical staining (IHC) in a prospectively collected, population-based cervical cancer cohort of 508 patients with corresponding clinicopathological and follow-up data. Staining was scored as either negative or positive and was further defined by the staining index (SI), consisting of area and intensity of staining varying from 1–9. MMR deficiency was defined as negative expression in one or more of the proteins. Also, gene set enrichment analyses were performed and differentially expressed or mutated genes were identified, across the RNA and whole-exome sequencing cohorts (n=72 and n=75, respectively), consisting of data obtained from fresh tissue. Results Eight tumours (1.5%) were MMR deficient, four of which were of neuroendocrine histology. MMR status did not independently correlate with survival when adjusted for histologic type (HR 1.93, p=0.222). Low MSH-2 (SI ≤4, n=48) associated with poor survival (HR: 1.94 p=0.02), also when corrected for tumour stage, type and patient age (HR 2.06, p=0.013). Furthermore, the MSH-2 low tumours associated with high tumour mutational burden (p=0.003) and a high frequency of (frameshift) mutations in the double-strand break repair gene RAD50 (p<0.001). Conclusion MMR deficiency is rare in cervical cancer and exhibits no independent relationship to survival in the current study. Low MSH-2 level is an independent prognostic marker for poor survival in cervical cancer.

Background The endometrial cancer mismatch repair (MMR) deficient subgroup is defined by loss of MSH6, MSH2, PMS2 or MLH1. We compare MMR status in paired preoperative and operative samples and investigate the prognostic impact of differential MMR protein expression levels. Methods Tumour lesions from 1058 endometrial cancer patients were immunohistochemically stained for MSH6, MSH2, PMS2 and MLH1. MMR protein expression was evaluated as loss or intact to determine MMR status, or by staining index to evaluate the prognostic potential of differential expression. Gene expression data from a local ( n = 235) and the TCGA ( n = 524) endometrial cancer cohorts was used for validation. Results We identified a substantial agreement in MMR status between paired curettage and hysterectomy samples. Individual high expression of all four MMR markers associated with non-endometrioid subtype, and high MSH6 or MSH2 strongly associated with several aggressive disease characteristics including high tumour grade and FIGO stage, and for MSH6, with lymph node metastasis. In multivariate Cox analysis, MSH6 remained an independent prognostic marker, also within the endometrioid low-grade subgroup ( P < 0.001). Conclusion We demonstrate that in addition to determine MMR status, MMR protein expression levels, particularly MSH6, may add prognostic information in endometrial cancer.

Advanced cervical cancer carries a particularly poor prognosis, and few treatment options exist. Identification of effective molecular markers is vital to improve the individualisation of treatment. We investigated transcriptional data from cervical carcinomas related to patient survival and recurrence to identify potential molecular drivers for aggressive disease.Primary tumour RNA-sequencing profiles from 20 patients with recurrence and 53 patients with cured disease were compared. Protein levels and prognostic impact for selected markers were identified by immunohistochemistry in a population-based patient cohort.Comparison of tumours relative to recurrence status revealed 121 differentially expressed genes. From this gene set, a 10-gene signature with high prognostic significance (p = 0.001) was identified and validated in an independent patient cohort (p = 0.004). Protein levels of two signature genes, HLA-DQB1 (n = 389) and LIMCH1 (LIM and calponin homology domain 1) (n = 410), were independent predictors of survival (hazard ratio 2.50, p = 0.007 for HLA-DQB1 and 3.19, p = 0.007 for LIMCH1) when adjusting for established prognostic markers. HLA-DQB1 protein expression associated with programmed death ligand 1 positivity (p < 0.001). In gene set enrichment analyses, HLA-DQB1high tumours associated with immune activation and response to interferon-γ (IFN-γ).This study revealed a 10-gene signature with high prognostic power in cervical cancer. HLA-DQB1 and LIMCH1 are potential biomarkers guiding cervical cancer treatment.

Obesity is a disease characterized by chronic low-grade systemic inflammation and has been causally linked to the development of 13 cancer types. Several studies have been undertaken to determine whether tumors evolving in obese environments adapt differential interactions with immune cells and whether this can be connected to disease outcome. Most of these studies have been limited to single-cell lines and tumor models and analysis of limited immune cell populations. Given the multicellular complexity of the immune system and its dysregulation in obesity, we applied high-dimensional suspension mass cytometry to investigate how obesity affects tumor immunity. We used a 36-marker immune-focused mass cytometry panel to interrogate the immune landscape of orthotopic syngeneic mouse models of pancreatic and breast cancer. Unanchored batch correction was implemented to enable simultaneous analysis of tumor cohorts to uncover the immunotypes of each cancer model and reveal remarkably model-specific immune regulation. In the E0771 breast cancer model, we demonstrate an important link to obesity with an increase in two T-cell-suppressive cell types and a decrease in CD8 T cells.

Obesity is a disease characterized by chronic low-grade systemic inflammation and has been causally linked to the development of 13 cancer types. Several studies have been undertaken to determine if tumors evolving in obese environments adapt differential interactions with immune cells and if this can be connected to disease outcome. Most of these studies have been limited to single cell lines and tumor models and analysis of limited immune cell populations. Given the multicellular complexity of the immune system and its dysregulation in obesity, we applied high-dimensional suspension mass cytometry to investigate how obesity affects tumor immunity. We used a 36-marker immune-focused mass cytometry panel to interrogate the immune landscape of orthotopic syngeneic mouse models of pancreatic and breast cancer. Unanchored batch correction was implemented to enable simultaneous analysis of tumor cohorts to uncover the immunotypes of each cancer model and reveal remarkably model-specific immune regulation. In the E0771 breast cancer model, we demonstrate an important link to obesity with an increase in two T cell suppressive cell types and a decrease in CD8 T-cells.

Objective PD-L1 and PD-1 are predictive markers for immunotherapy and increasingly relevant in endometrial cancer. The reported fraction of positive primary tumors has been inconsistent. We investigated the expression of PD-L1 and PD-1 in primary tumors, also stratified by MSI. As immunotherapy is foremost relevant for metastatic disease, PD-L1 and PD-1 expression was also assessed in corresponding metastatic lesions. Methods PD-L1 and PD-1 was investigated in a prospective, population based endometrial cancer cohort of 700 patients with corresponding metastatic lesions from 68 and 74 patients respectively. Fresh tissue was used for gene expression analysis. Results In primary tumors, PD-L1 and PD-1 are expressed in 59% and 63%, respectively, but with no impact on survival, nor when stratified for MSS and MSI. Expression patterns of PD-L1 and PD-1 are similar in MSI and MSS tumors. Available metastatic lesions show heterogeneous expression of PD-L1 and PD-1. In gene expression analysis several genes related to immunological activity, including CD274 (encoding for PD-L1), were upregulated in PD-1 positive tumors. Conclusion PD-L1 and PD-1 are frequently expressed in endometrial cancer and expression patterns are similar across MSS and MSI tumors. Expression in corresponding metastatic lesions is discordant compared to primary tumors. These findings are in particular relevant for treatment decisions in advanced and recurrent disease.

BACKGROUND:Bayesian networks (BNs) are machine-learning-based computational models that visualize causal relationships and provide insight into the processes underlying disease progression, closely resembling clinical decision-making. Preoperative identification of patients at risk for lymph node metastasis (LNM) is challenging in endometrial cancer, and although several biomarkers are related to LNM, none of them are incorporated in clinical practice. The aim of this study was to develop and externally validate a preoperative BN to predict LNM and outcome in endometrial cancer patients. METHODS AND FINDINGS:Within the European Network for Individualized Treatment of Endometrial Cancer (ENITEC), we performed a retrospective multicenter cohort study including 763 patients, median age 65 years (interquartile range [IQR] 58-71), surgically treated for endometrial cancer between February 1995 and August 2013 at one of the 10 participating European hospitals. A BN was developed using score-based machine learning in addition to expert knowledge. Our main outcome measures were LNM and 5-year disease-specific survival (DSS). Preoperative clinical, histopathological, and molecular biomarkers were included in the network. External validation was performed using 2 prospective study cohorts: the Molecular Markers in Treatment in Endometrial Cancer (MoMaTEC) study cohort, including 446 Norwegian patients, median age 64 years (IQR 59-74), treated between May 2001 and 2010; and the PIpelle Prospective ENDOmetrial carcinoma (PIPENDO) study cohort, including 384 Dutch patients, median age 66 years (IQR 60-73), treated between September 2011 and December 2013. A BN called ENDORISK (preoperative risk stratification in endometrial cancer) was developed including the following predictors: preoperative tumor grade; immunohistochemical expression of estrogen receptor (ER), progesterone receptor (PR), p53, and L1 cell adhesion molecule (L1CAM); cancer antigen 125 serum level; thrombocyte count; imaging results on lymphadenopathy; and cervical cytology. In the MoMaTEC cohort, the area under the curve (AUC) was 0.82 (95% confidence interval [CI] 0.76-0.88) for LNM and 0.82 (95% CI 0.77-0.87) for 5-year DSS. In the PIPENDO cohort, the AUC for 5-year DSS was 0.84 (95% CI 0.78-0.90). The network was well-calibrated. In the MoMaTEC cohort, 249 patients (55.8%) were classified with <5% risk of LNM, with a false-negative rate of 1.6%. A limitation of the study is the use of imputation to correct for missing predictor variables in the development cohort and the retrospective study design. CONCLUSIONS:In this study, we illustrated how BNs can be used for individualizing clinical decision-making in oncology by incorporating easily accessible and multimodal biomarkers. The network shows the complex interactions underlying the carcinogenetic process of endometrial cancer by its graphical representation. A prospective feasibility study will be needed prior to implementation in the clinic.

ObjectiveBetter biomarkers are needed in order to identify patients with endometrial carcinoma at risk of recurrence and who may profit from a more aggressive treatment regimen. Our objective was to explore the applicability of plasma growth differentiation factor 15 (GDF-15) as a marker for recurrent disease, as well as a marker for poor prognosis and lymph node metastases.MethodsEDTA-blood samples were obtained from 235 patients with endometrial cancer before primary surgery. For 36 of these patients, matching blood samples were collected at time of recurrence. Blood samples were also collected from 78 patients with endometrial hyperplasia. Plasma GDF-15 was measured by an enzyme-linked immunosorbent assay (ELISA). Preoperative pelvic MRI scans for 141 patients were investigated in parallel for imaging variables.ResultsPreoperative plasma level of GDF-15 was significantly higher for patients who experienced recurrence (1780 ng/L; 95% CI; 518-9475 ng/L) than for patients who did not develop recurrent disease (1236 ng/L; 95% CI; 307-7030 ng/L) (p<0.001). Plasma levels of GDF-15 at recurrence (2818 ng/L, 95% CI 2088-3548 ng/L) were significantly higher than plasma levels of GDF-15 measured at time of primary diagnosis (1857 ng/L, 95% CI; 1317-2398 ng/L) (p = 0.001). High plasma level GDF-15 independently predicts recurrent disease (OR = 3.14; 95% CI 2.10-4.76) and lymph node metastases (OR = 2.64; 95% CI 1.52-4.61). Patients with high plasma level of GDF-15 had significantly larger tumor volume (p = 0.008).ConclusionElevated plasma level of GDF-15 is associated with aggressive disease and lymph node metastasis in endometrial carcinoma. GDF-15 may be helpful in indicating recurrent disease.

BACKGROUND:Cervical cancer is a major health problem worldwide. Identification of effective clinicopathologic and molecular markers is vital to improve treatment stratification. OBJECTIVES:The purpose of this study was to validate a set of well-defined clinicopathologic features in a large population-based, prospectively collected cervical cancer cohort to support their use in the clinic. Further, we explored p53 and human epidermal growth factor receptor 2 as potential prognostic markers in cervical cancer. STUDY DESIGN:Tissue was collected from 401 patients with cervical cancer. Clinical data that included follow-up evaluations were collected from patient journals. Histopathologic data were evaluated and revised by an expert pathologist. The prognostic impact of selected clinicopathologic variables was analyzed in the whole cohort. Tissue microarrays were prepared from 292 carcinomas, and p53 and human epidermal growth factor receptor 2 protein levels were evaluated by immunohistochemistry. Fresh frozen samples from overlapping cervical carcinomas previously were subjected to human papilloma virus typing (n=94), whole exome (n=100) and RNA (n=79) sequencing; the results were available for our analyses. RESULTS:Among the clinicopathologic variables, vascular space invasion, histologic type, and tumor size were verified as strong independent prognostic markers. High p53 protein levels were associated significantly with markers for aggressive phenotype and survival, also in multivariate survival analysis, but did not reflect TP53 mutational status. High human epidermal growth factor receptor 2 protein levels were identified in 21% of all tumors. ERBB2 amplification was associated with poor outcome (P=.003); human epidermal growth factor receptor 2 protein level was not. CONCLUSIONS:Our findings support that the Féderation Internationale de Gynécologie et d'Obstétrique s guidelines should include vascular space invasion and tumor size 2-4 cm and that careful selection of histologic type is essential for stratification of patient risk groups. High p53 levels independently predict poor survival yet do not reflect mutational status in cervical cancer. Amplified ERBB2 significantly links to poor survival, while HercepTest does not. With optimal stratification, human epidermal growth factor receptor 2-based therapy may improve cervical cancer treatment.

Abstract Cervical cancer is the third leading cause of cancer in the female population worldwide, causing the death of more than 240,000 patients annually in developing countries. Increased molecular knowledge is crucial to identify robust prognostic and predictive biomarkers that can better guide treatment. The tumor response to trastuzumab is well established and strongly links to HER2 expression status evaluated by the Hercep Test, hence, it is essential to define the level of expression of this receptor using the FDA-approved Hercep Test to stratify cervical cancer patients with potential benefits from trastuzumab treatment. Comprehensive molecular characterization has been conducted on 88 paired normal and tumor cases identifying ERBB2 to be frequently altered in cervical cancers. We here explore the protein expression of HER2 by immunohistochemical staining in a larger validation series (n = 220) and relate HER2 expression to the ERBB2 gene alterations, patients molecular profile and clinicopathological features. We find a highly significant correlation between Hercep Test score and mRNA ERBB2 expression (p<0.001). The level of ERBB2 mRNA was also significantly associated with copy number status (p = 0.007). Further clinocopathological parameters like high FIGO stage, high grade, adenocarcinomas and normal p53 status was significantly linked to high HER2 protein expression. Kaplan Meier survival analysis revealed that within the squamous cell carcinomas, high protein levels of HER2 was linked to poorer disease specific survival. Our results show a link between ERBB2 amplification, high mRNA expression and protein levels for HER2 in aggressive cervical cancers. Further studies of HER2 as a potential predictive marker for response to trastuzumab treatment in cervical cancer are needed. Citation Format: Mari K. Halle, Akinyemi I. Ojesina, Ingvild L. Tangen, Frederik Holst, Hilde R. Engerud, Bjørn I. Bertelsen, Camilla Krakstad, Helga B. Salvesen. HER2 as a potential predictive marker and target for therapy in cervical cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr LB-120. doi:10.1158/1538-7445.AM2015-LB-120

Abstract Background: The transcriptional regulator heat-shock transcription factor 1 (HSF1) has recently been linked to progression of breast cancer (Santagata, PNAS 2011). HSF1 is a regulator of cellular response to stress and drives the production of heat shock proteins (HSPs). The role of HSF1 to promote tumor cell survival has been demonstrated in hsf1 knockout mice (Dai, Cell 2007). Furthermore, recent identification of a specific role of HSF1 in cancer progression has led to new relevance of HSF1 as a marker both for prognosis and as a predictive marker. The role of HSF1 in endometrial cancer has so far been unexplored. Methods: A total of 823 lesions from endometrial carcinoma precursors, primary tumors and metastases were prospectively collected and explored for HSF1 protein expression in relation to established markers for aggressive disease and survival. Transcriptional alterations related to HSF1 protein level were investigated by microarray analysis for 224 freshly frozen samples in parallel. Results: High expression of HSF1 protein in endometrial carcinoma is significantly associated with non-endometrioid histology, high grade, ERα loss and poor survival (all p-values ≤0.02). Also among ERα-positive patients presumed to have good prognosis, high expression of HSF1 is correlated with poor survival (p=0.02). HSF1 mRNA and protein levels increase significantly from primary tumors to metastases (p<0.03). HSF1-related gene signatures were explored. Such signatures increase from complex atypical hyperplasias through primary tumors to metastatic lesion and were also found to have prognostic value. HSF1 immunohistochemistry suggests the use of HSP90 inhibitiors as potential novel therapeutic approach in cases with high protein expression of HSF1. Conclusion: We demonstrate for the first time in endometrial cancer that high expression of HSF1 and measures for transcriptional activation of HSF1 associates with poor outcome, progression from precursor lesions through primary tumors to metastatic lesions. Also, HSP90 inhibitors are suggested as potential new targeted therapeutics for cases with high HSF1 levels in particular. 1. Santagata,S., et al., High levels of nuclear heat-shock factor 1 (HSF1) are associated with poor prognosis in breast cancer. Proc Natl Acad Sci U S A, 2011. 108(45):p. 18378-83. 2. Dai,C., et al., Heat shock factor 1 is a powerful multifaceted modifier of carcinogenesis. Cell, 2007. 130(6): p. 1005-18. Citation Format: Hilde Engerud, Ingvild Løberg Tangen, Anna Berg, Kanthida Kusonmano, Ingunn Stefansson, Helga B. Salvesen, Camilla Krakstad. High level of nuclear heat-shock factor 1 is associated with aggressive disease and suggests targets for therapy in endometrial carcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4731. doi:10.1158/1538-7445.AM2014-4731

Many emerging tools for comprehensive molecular profiling of malignant lesions demand fresh frozen tissue with a high tumor purity. Often, a tumor epithelial content of at least 80% is recommended. This approach may lead to a systematic bias, and therefore we explore if this introduces a selection of cases with a certain phenotype in cervical cancer. Clinicopathologic data for a population-based cohort of 328 patients have been studied. Fresh frozen tumor specimens were available for 151 of these patients and investigated for epithelial tumor cell portion in hematoxylin-stained frozen sections by light microscopy. The estimated tumor purity in the samples was compared with FIGO stage, histopathologic characteristics and survival. High tumor purity was significantly more often found in squamous cell carcinoma (SCC) compared to adenocarcinoma (AC) (P = 0.03). For the subgroup of AC (n = 40), there was a significant association between high tumor purity in the fresh frozen samples and later occurrence of recurrent disease (P = 0.04). In SCC, no significant associations between tumor purity and disease stage, grade or outcome were found. Apparently in line with this, grade was found to influence prognosis in AC, but not in SCC. Our findings suggest that selection of samples based on high tumor purity in fresh frozen tissue may introduce a selection bias toward aggressive disease for the subgroup of AC, but not for SCC of the cervix. Thus, the prevalence of potential molecular biomarkers identified in AC in particular should be validated in a population-based setting to further explore clinical relevance. Also, molecular biomarkers only prevalent in subgroups with low tumor purity may go undetected in sample collections enriched for high tumor purity.

Objectives. The objective of this study is to investigate preoperative hematological parameters for anemia, leukocytosis and thrombocytosis in relation to established prognostic factors and survival in endometrial cancer.Methods. 557 patients treated for endometrial carcinoma were prospectively included in a study focusing on the relationship between preoperative hemoglobin, leukocyte and platelet counts, and a panel of clinicopathological characteristics and outcome.Results. Preoperative anemia was present in 15.8%, leukocytosis in 11.2% and thrombocytosis in 12.1%. Among patients with localized disease (FIGO stage I/II), 18.1% had anemia and/or thrombocytosis at diagnosis. Patients with advanced disease (high FIGO stage and lymph-node metastasis) had significantly lower hemoglobin count, higher leukocyte count and higher platelet count (all p < 0.008). Patients with anemia, leukocytosis and thrombocytosis had significantly shorter 5-year disease-specific survival of 61.3%, 66.0% and 61.0% respectively, compared to 87.7%, 86.3% and 87.3% for patients with normal counts (all p < 0.001). In multivariate Cox regression analysis, lower hemoglobin counts and higher platelet counts were independently associated with poor outcome adjusted for the standardly applied prognostic markers (p < 0.033).Conclusion. Preoperative anemia, leukocytosis or thrombocytosis in women with endometrial carcinoma is associated with advanced disease and poor disease-specific survival. (C) 2013 Elsevier Inc. All rights reserved.