Improving endometrial carcinoma diagnostics

Endometrial carcinoma is the most common gynecological malignancy and greatly affects global disease burden. Despite it being among the most common cancers it is relatively understudied and until recently poorly understood. Diagnostics of endometrial carcinoma have been underdeveloped for many years resulting in perpetual poor interobserver reproducibility in turn inhibiting correct diagnosis and complicating comparisons of clinical trials. In this thesis we used a three-tiered approach in trying to improve endometrial carcinoma diagnostics: strictly improving morphological criteria, adding immunohistochemistry (IHC)/biomarker panels, as well as genomics data in order to improve diagnostic and prognostic ability. In study 1 we compared standard FIGO grading of endometrial carcinoma to a newly developed morphologic system we named cell-type independent (CTI). We compared interobserver reproducibility using both systems in 70 patient`s endometrial biopsies. We could not identify any significant improvement in the form of increased interobserver reproducibility nor increased accuracy when compared to final hysterectomy diagnosis. We conclude that the limitations of the biopsy setting hampered the CTI system`s ability to outperform current FIGO grading system and that the assessment of nuclear grade, no matter what the definitions of a high nuclear grade are, remain subjective. In study 2 12 separate subspecialized gynaecological pathologists reviewed pathology slides of 70 cases of endometrial biopsies. We compared the interobserver agreement when adding a biomarker panel (IHC for p53, ER and PGR and DNA ploidy analysis) to only standard morphology diagnosis with accompanying results of final hysterectomy diagnosis. The addition of a biomarker panel increased interobserver agreement from 75,8%, kappa = 0.52 to 84%, kappa= 0.68. Diagnostic agreement to final hysterectomy diagnosis also increased with use of the panel, going from 83.6% to 88.7% agreement with final diagnosis after incorporating the panel in biopsy diagnosis (p<0.05). The two biomarkers p53 IHC and DNA ploidy analysis showed significant effect on up-or downgrading of tumors. We conclude that selective use of a simple biomarker panel greatly aids in endometrial biopsy diagnostics. Study 3 was a collaborative effort between Bern and Karolinska University Hospital we sought to implement a surrogate molecular marker in 604 patients with endometrial carcinoma in order to try to replicate the TCGA (The cancer genome atlas) molecular markers classifiers and to hopefully aid in endometrial carcinoma diagnostics and prognostics. The panel consisted of IHC for p53 (to define a p53mut group) and MSH2, MSH6, PMS2, MLH1 (to define a MMRd, mismatch repair defective, group) and Sanger sequencing of the POLE gene (to define a POLEmut) group. We found that this simple surrogate molecular marker, using readily available methods, could reproduce the findings of the TCGA and render four prognostically different molecular categories. We could however not prove an added prognostic benefit over current risk assessment prognostic tools, nor any benefit in adding a surrogate molecular marker to current prognostic markers. Moreover only one of the molecular groups (p53 mut) proved to be statistically significant marker for predicting Progression free survival (PFS) and overall survival (OS). From this we concluded that in the setting of non-selected group of patients consisting mostly of low grade tumors proving significant differences between each of the molecular groups couldn’t be done since progression events were so rare and the group as a whole behaved indolently. Nonetheless there are clear benefits with the addition of genomic data for other purposes such as aid in suitable patient selection for adjuvant treatment. In study 4 we conducted sequencing of exons 9-14 of the POLE gene in a cohort of 604 patients with endometrial carcinoma in order to more deeply analyze mutation events and the phenotype of POLE mutated endometrial carcinomas. We found mostly previously described hotspot mutations but also a substantial number of mutations with unknown significance and characterized POLEmut tumors as occurring in mainly younger women with nulliparity and consisting predominantly of endometrioid endometrial carcinomas. This study shows that when POLE mutations are clearly defined as either hotspot mutations and/or POLE mutations with a known hypermutated phenotype this patient group of POLEmut had very good outcomes with only 1 recurrence in 38 patients. The Cox regression couldn’t however show any significant difference between POLEmut and non POLEmut tumors, however we believe again this was because we are dealing with a low risk EC population where both recurrence events and POLE mutations are rare, and thus we believe we were underpowered

Retrospective evaluation of whole exome and genome mutation calls in 746 cancer samples

Funder: NCI U24CA211006Abstract: The Cancer Genome Atlas (TCGA) and International Cancer Genome Consortium (ICGC) curated consensus somatic mutation calls using whole exome sequencing (WES) and whole genome sequencing (WGS), respectively. Here, as part of the ICGC/TCGA Pan-Cancer Analysis of Whole Genomes (PCAWG) Consortium, which aggregated whole genome sequencing data from 2,658 cancers across 38 tumour types, we compare WES and WGS side-by-side from 746 TCGA samples, finding that ~80% of mutations overlap in covered exonic regions. We estimate that low variant allele fraction (VAF < 15%) and clonal heterogeneity contribute up to 68% of private WGS mutations and 71% of private WES mutations. We observe that ~30% of private WGS mutations trace to mutations identified by a single variant caller in WES consensus efforts. WGS captures both ~50% more variation in exonic regions and un-observed mutations in loci with variable GC-content. Together, our analysis highlights technological divergences between two reproducible somatic variant detection efforts

Phenotype of POLE-mutated endometrial cancer.

Background and purposeIndividualized therapy in endometrial cancer, the most common gynaecologic cancer in the developed world, focuses on identifying specific molecular subtypes. Mutations in the exonuclease domain of the DNA polymerase epsilon (POLE) gene define one such subtype, which causes an ultra-mutated tumour phenotype. These tumours may have an improved progression-free survival and may be receptive to specific therapies. However, the clinical phenotype of these tumours is unknown. The objective of this study was to evaluate the clinical and genetic features of POLE-mutated tumours from a large cohort of women whose cases are characterized by: (1) the availability of detailed clinical and lifestyle data; (2) mutation analysis; and (3) long-term follow-up.MethodsA total of 604 patients with endometrial cancer were included in the study. Data from a detailed questionnaire, including lifestyle and family history information, provided extensive pertinent information on the patients. Sequencing of exons 9-14 of the POLE gene was performed. Follow-up data were gathered and analysed.ResultsHotspot pathogenic POLE mutations were identified in N = 38/599 patients (6.3%). Patients with a POLE-mutated tumour were significantly younger, were more often nulliparous, and had a history of smoking. POLE-mutated tumours were more frequently aneuploid. Prognosis for patients with hotspot POLE-mutated tumours was significantly better in comparison with patients with non-mutated tumours; however careful selection of pathogenic mutations is essential to the definition of this prognostically favourable group.ConclusionsThis study demonstrates that POLE-mutated endometrial cancer is significantly associated with previously unknown clinicopathologic characteristics. Outcome in POLE-mutated tumours was excellent in cases with hotspot mutations. Our results suggest that prediction of excellent outcome in cases of POLE-mutated EMCA should be restricted to cases of EMCA with hotspot mutations until further data are available on the rising number of mutations with unknown significance

Implementation of the 2021 molecular ESGO/ESTRO/ESP risk groups in endometrial cancer.

INTRODUCTION In 2021, a joint ESGO/ESTRO/ESP committee updated their evidence-based guidelines for endometrial cancer, recommending a new risk grouping incorporating both clinicopathologic and molecular parameters. We applied the new risk grouping and compared the results to those of the prior 2016 clinicopathologic system. MATERIALS AND METHODS We classified molecularly a cohort of 604 women diagnosed with endometrial cancer using immunohistochemistry for TP53 and MMR proteins on a tissue microarray, as well as Sanger sequencing for POLE mutations. These results, combined with clinicopathologic data, allowed the patients to be risk grouped using both the new 2021 molecular/clinicopathologic parameters and the prior 2016 clinicopathologic system. RESULTS The application of the 2021 molecular markers shows Kaplan-Meier curves with a significant difference between the groups for all survival. Molecular classification under the 2021 guidelines revealed a total of 39 patients (39/594, 7%) with a change in risk group in relation to the 2016 classification system: the shift was alone due to either P53abn or POLEmut molecular marker. In order to ensure correct 2021 molecular risk classification, not all patients with endometrial cancer need a molecular diagnostic: 433 (72.9%) cases would need to be analyzed by TP53 IHC, only 46 (7.7%) by MMR IHC and 286 (48.1%) POLE sequencing reactions. CONCLUSION Application of the 2021 molecular risk groups is feasible and shows significant differences in survival. IHC for TP53 and MMR and applying POLE sequencing is only needed in selected cases and leads to shifting risk groups both upward and downward for a sizeable number of patients. It is possible to significantly reduce the number of analyses required to implement the classification if resources are limited

A Selective Biomarker Panel Increases the Reproducibility and the Accuracy in Endometrial Biopsy Diagnosis

Grading and histologic typing of endometrial cancer in biopsy material has a direct impact on the decision to perform lymphadenectomy and/or omentectomy in many cancer centers. Endometrial biopsies are among the most common general surgical pathology specimens. Multiple studies have shown that biopsy diagnosis suffers from a lack of reproducibility. Although many biomarkers have been proposed, none have been demonstrated to improve the diagnosis in the biopsy setting. In this study, 70 biopsies with endometrial carcinoma were supplemented with a biomarker panel consisting of ER, PR, P53, and DNA ploidy. A representative H&E slide was scanned digitally and made available to 12 gynecologic pathologists in 4 Nordic countries: Finland, Denmark, Sweden, and Norway. Reviewers diagnosed the cases both before and after being provided with the biomarker results. The interobserver percent agreement and Cohen κ improved from 75.8% (κ=0.52, moderate) to 84% (κ=0.68, substantial) with inclusion of the biomarker panel. Agreement with the subsequent hysterectomy diagnosis also improved from 83.6% (κ=0.67) to 88.7% (κ=0.77). There was no statistical improvement between a reflex (84% agreement) and a reflective testing algorithm (82.9% agreement), suggesting that the selective use of biomarkers is appropriate. Difficult cases were almost exclusively high-grade tumors. Finally, a statistical model indicated that only P53 and DNA ploidy, in conjunction with an H&E review, had an impact on the decision to upgrade or downgrade cases

Clinical and Ultrasound Characteristics of the Microcystic Elongated and Fragmented (MELF) Pattern in Endometrial Cancer According to the International Endometrial Tumor Analysis (IETA) criteria

OBJECTIVES: To describe sonographic features of the microcystic elongated and fragmented (MELF) pattern of myometrial invasion (MI) using the International Endometrial Tumor Analysis (IETA) criteria; to assess the effect of the MELF pattern on preoperative ultrasound evaluation of MI; and to determine the relationship of the MELF pattern to more advanced stage ( 65 IB) and lymph node metastases (LNM) in women with endometrioid endometrial cancer (EEC). METHODS/MATERIALS: We included 850 women with EEC from the prospective IETA 4 study. Ultrasound experts performed all ultrasound examinations, accordingto the IETA protocol. Reference pathologists assessed the presence orabsence of the MELF pattern. Sonographic features and accuracy of ultrasound assessment of MI were compared in cases with the presence and the absence of the MELF pattern. The MELF pattern was correlated to more advanced stage ( 65IB) and LNM. RESULTS: The MELF pattern was present in 197 (23.2%) women. On preoperative ultrasound imaging the endometrium was thicker (p = 0.031), more richly vascularized (p = 0.003) with the multiple multifocal vessel pattern (p < 0.001) and the assessment of adenomyosis was more often uncertain (p < 0.001). The presence or the absence of the MELF pattern did not affect the accuracy of the assessment of MI. The MELF pattern was associated with MI 65 50% (p < 0.001), cervical stromal invasion (CSI) (p = 0.037), more advanced stage ( 65 IB) (p < 0.001) and LNM (p = 0.011). CONCLUSIONS: Tumors with the MELF pattern were slightly larger, more richly vascularized with multiple multifocal vessels and assessment of adenomyosis was more uncertain on ultrasound imaging. The MELF pattern did not increase the risk of underestimating MI in preoperative ultrasound staging. Tumors with the MELF pattern were more than twice as likely to have more advanced stage ( 65 IB) and LNM

Advanced Modeling and Simulation Tools to Address Build-Up Issues in Additive Manufacturing by Cold Spray

International audienc

Sex differences in oncogenic mutational processes

Funder: Canadian Network for Research and Innovation in Machining Technology, Natural Sciences and Engineering Research Council of Canada (NSERC Canadian Network for Research and Innovation in Machining Technology); doi: https://doi.org/10.13039/501100002790Funder: Genome Canada (Génome Canada); doi: https://doi.org/10.13039/100008762Funder: Canada Foundation for Innovation (Fondation canadienne pour l'innovation); doi: https://doi.org/10.13039/501100000196Funder: Terry Fox Research Institute (Institut de Recherche Terry Fox); doi: https://doi.org/10.13039/501100004376Abstract: Sex differences have been observed in multiple facets of cancer epidemiology, treatment and biology, and in most cancers outside the sex organs. Efforts to link these clinical differences to specific molecular features have focused on somatic mutations within the coding regions of the genome. Here we report a pan-cancer analysis of sex differences in whole genomes of 1983 tumours of 28 subtypes as part of the ICGC/TCGA Pan-Cancer Analysis of Whole Genomes (PCAWG) Consortium. We both confirm the results of exome studies, and also uncover previously undescribed sex differences. These include sex-biases in coding and non-coding cancer drivers, mutation prevalence and strikingly, in mutational signatures related to underlying mutational processes. These results underline the pervasiveness of molecular sex differences and strengthen the call for increased consideration of sex in molecular cancer research