Clinical evaluation of a low-coverage whole-genome test for detecting homologous recombination deficiency in ovarian cancer.

BACKGROUND: The PAOLA-1/ENGOT-ov25 trial showed that maintenance olaparib plus bevacizumab increases survival of advanced ovarian cancer patients with homologous recombination deficiency (HRD). However, decentralized solutions to test for HRD in clinical routine are scarce. The goal of this study was to retrospectively validate on tumor samples from the PAOLA-1 trial, the decentralized SeqOne assay, which relies on shallow Whole Genome Sequencing (sWGS) to capture genomic instability and targeted sequencing to determine BRCA status. METHODS: The study comprised 368 patients from the PAOLA-1 trial. The SeqOne assay was compared to the Myriad MyChoice HRD test (Myriad Genetics), and results were analyzed with respect to Progression-Free Survival (PFS). RESULTS: We found a 95% concordance between the HRD status of the two tests (95% Confidence Interval (CI); 92%-97%). The Positive Percentage Agreement (PPA) of the sWGS test was 95% (95% CI; 91%-97%) like its Negative Percentage Agreement (NPA) (95% CI; 89%-98%). In patients with HRD-positive tumors treated with olaparib plus bevacizumab, the PFS Hazard Ratio (HR) was 0.38 (95% CI; 0.26-0.54) with SeqOne assay and 0.32 (95% CI; 0.22-0.45) with the Myriad assay. In patients with HRD-negative tumors, HR was 0.99 (95% CI; 0.68-1.42) and 1.05 (95% CI; 0.70-1.57) with SeqOne and Myriad assays. Among patients with BRCA-wildtype tumors, those with HRD-positive tumors, benefited from olaparib plus bevacizumab maintenance, with HR of 0.48 (95% CI: 0.29-0.79) and of 0.38 (95% CI: 0.23 to 0.63) with the SeqOne and Myriad assay. CONCLUSION: The SeqOne assay offers a clinically validated approach to detect HRD.

[The "Conseil national professionnel d'anatomie et cytologie pathologiques" (CNPath): At the service of the profession]. / Le Conseil national professionnel d'anatomie et cytologie pathologiques (CNPath) : une structure au service de la profession.

The « conseils nationaux professionnels ¼ (CNP) are professional boards existing since 2010. Their missions, organization and functioning have been defined by the decree 2019-17 of January 9, 2019. CNPs represent all the members of a medical specialty (or health profession). CNPs must include all the learned societies and all the representative structures and associations of the same medical specialty. Their bodies must strictly respect the parity between public and private health sectors. The main missions of CNPs include the contribution to the elaboration of the national priority directions for continuous medical education and the definition of the individual plan for continuous professional development (DPC) recommended for the specialty. CNPs also behave as a single window for ministries, State agencies, welfare system and colleges of physicians. They are likely to be strongly involved in the process of re-certification of physicians, established in July 2019. The Conseil national professionnel d'anatomie et cytologie pathologiques, termed CNPath, has been created in 2010 and officially recognized by the Ministry of Health in August 2019. The main current actions of CNPath are: the elaboration of the individual DPC scheme for the specialty and the definition of the minimal obligations requested for its validation, the long-expected recognition of the expertal consultation in pathology, the support to the nation-wide effort for the production of structured pathological reports and the launching of a plan for implementing digital pathology. An internet site is under construction, to diffuse all the relevant information and make available the documents useful to all pathologists.

HER2 gene and protein expression status of breast carcinoma can be reliably tested on a single slide.

Human epidermal growth factor receptor 2 (HER2) status in breast carcinomas serves as a predictor of benefit from anti-HER2 therapy. In providing clinicians with the information necessary to decide whether or not to treat with targeted therapy, it might be necessary to choose between methods assessing HER2 protein overexpression or gene amplification. A new diagnostic approach could be a combination of both tests on the same slide. If accurate and reproducible, this approach might optimize patient stratification for therapy. In this study, formalin-fixed paraffin-embedded tumor samples from 77 breast cancer patients were examined for HER2 by immunohistochemistry (IHC) and silver in situ hybridization (SISH) using HER2 IHC (clone 4B5), HER2/CEN17SISH, and combined IHC and SISH assay, called gene protein (GP). Cases were selected to ensure a sufficient number of borderline cases on the basis of IHC scores (0, 1+, 2+, 3+), obtained during diagnostic histopathological workup. The concordance between the HER2 IHC score obtained during diagnostic histopathological workup and GP was 93 %. Discordances had no influence on therapy decisions. The concordance between ISH results using dual ISH (DISH) and GP was 96 %. Of the 77 cases studied by GP, three cases with a ratio close to 2 would have been called amplified by DISH. The use of GP reduced the time for slide reading for a trained pathologist by up to 25 %, relative to sequential reading of IHC followed by SISH. For cases with an IHC score of 2+, the final result was obtained in 1 day, while the sequential technique would have required retesting by ISH on a second day. In conclusion, assessment of HER2 status by GP is an improvement for pathologists and facilitates clinical decision-making for breast cancer management.

[Pertuzumab and solid tumors: perspectives]. / Pertuzumab et tumeurs solides : perspectives.

Over the past decade, trastuzumab was the only available monoclonal anti-HER2 antibody for the treatment of HER2 positive breast and gastric cancer. Recently, pertuzumab added to docetaxel and trastuzumab showed dramatic overall survival improvement in first line treatment of HER2 positive metastatic breast cancer. Pertuzumab is the first approved monoclonal antibody in a new class of drugs called dimerization inhibitors. This agent was also approved in association with trastuzumab for neoadjuvant HER2-positive breast cancer treatment. However, pertuzumab development was not confined to breast cancer and in the present review, we will focus on biological rational, preclinical data and clinical trial results of pertuzumab in solid tumors excluding breast cancer.

CARM1 and PRMT1 are dysregulated in lung cancer without hierarchical features.

CARM1 and PRMT1 are 2 Protein Arginine Methyl Transferases (PRMT) dysregulated in cancer. CARM1 function is contradictory and depicted as facilitating proliferation or differentiation. PRMT1 is required for cell proliferation. CARM1 and PRMT1 cooperate for gene regulation. We report that CARM1 and PRMT1 are significantly overexpressed in 60 patients with Non-Small Cell Lung Carcinomas (NSCLC). CARM1 and PRMT1 correlated in healthy but not tumor tissue. Their levels of expression in tumor tissue were proportional to their levels of expression in the counterpart healthy tissue. Only CARM1 expression was found to be correlated with tumor differentiation and neither CARM1 nor PRMT1 expression was correlated with survival. Accordingly, CARM1 and PRMT1 are overexpressed in 2 NSCLC cell lines, A549 and H1299. Targeting PRMT1 with siRNA reduced proliferation, by decreasing cell growth and inhibiting soft agar colony formation, and promoted differentiation, by increasing the epithelial markers cytokeratin 7 and 8 and decreasing Neuromedin B receptor, which binds a mitogenic factor. siCARM1 yielded similar consequences but the conditions with siCARM1 reflected inhibition of both CARM1 and PRMT1. Together these results suggest that CARM1 and PRMT1 are involved in proliferation in lung cancer with no hierarchy of one protein over the other. The fact that CARM1 targeting suppresses PRMT1 in addition to CARM1 reinforces the functional importance of CARM1/PRMT1 interaction.

Submucosal plexitis as a predictor of postoperative surgical recurrence in Crohn's disease.

BACKGROUND: The presence of submucosal or myenteric plexitis was associated with clinical and endoscopic Crohn's disease (CD) recurrence after ileocolonic resection. We assessed the value of both submucosal and myenteric plexitis for predicting postoperative surgical recurrence in CD. METHODS: We performed a retrospective study using the database of the Department of Pathology of Nancy University Hospital. All patients who underwent CD-related resection between 1996 and 2008 were analyzed. The proximal resection margin was analyzed blindly by 2 expert pathologists. Plexitis was evaluated by counting each cell type (mast cell, plasmocyte, lymphocyte, eosinophil, and neutrophil) in both submucosal and myenteric plexuses. The optimal cut-off value for each cell type was determined by using receiver operating characteristic analysis. Cox proportional hazards regression analysis was used to identify independent predictors of the second CD-related surgery. RESULTS: Sixty-seven patients were included in the study. Median duration of follow-up was 46 months. Using Kaplan-Meier survival analysis, the proportion of patients without second surgery was 68% at 5 years. In multivariate analysis, using Cox proportional hazards regression analysis, early surgical revision after the first ileocecal resection (hazard ratio = 9.56; 95% confidence interval, 2.02-45.19; P = 0.0046), the presence of at least one eosinophil in the submucosal plexus (hazard ratio = 8.02; 95% confidence interval, 1.87-34.47; P = 0.0054), and the presence of more than 6 lymphocytes in the submucosal plexus (hazard ratio = 5.84; 95% confidence interval, 1.23-27.65; P = 0.0269) were independently associated with risk of surgical recurrence. CONCLUSIONS: Early surgical revision and submucosal plexitis in proximal margins of ileocolonic resection specimens are independently associated with CD surgical recurrence.

FVIIIra, CD15, and tryptase performance in the diagnosis of skin stab wound vitality in forensic pathology.

The timing of skin wounds is one of the most challenging problems in forensic pathology. In the first minutes or hours after infliction, histological examination fails to determine whether a wound was sustained before or after death. The aim of this study was to evaluate the use of three immunohistochemical markers (FVIIIra, CD15, and tryptase) for the interpretation of the timing of cutaneous stab wounds. We evaluated these markers in intravital wounds from autopsy cases (n = 12) and surgical specimens (n = 58). As controls, we used normal skin samples from autopsies (n = 8) and an original ex vivo surgical human model of recent postmortem wounds (n = 24). We found overexpression of FVIIIra in 100 % of vital wounds, but also in 53 % of the controls. The number of CD15-positive cells was higher in wound margins than in internal controls (p < 0.0001) and was significantly correlated with the time interval between incision and devascularization (p = 0.0005; minimal time for positivity, 9 min). Using the anti-tryptase antibody, we found that the mast cell degranulation rate was higher in wound margins (p < 0.0001) and correlated with the time interval (minimal time, 1 min). The sensitivity and specificity for the diagnosis of vitality were respectively 100 and 47 % for FVIIIra, 47 and 100 % for CD15, and 60 and 100 % for tryptase. The inter-observer agreement coefficients were 0.68 for FVIIIra, 0.90 for CD15, and 0.46 for tryptase. Finally, we demonstrated that these markers were not reliable in putrefied or desiccated specimens. In conclusion, CD15 and tryptase, but not FVIIIra, may be useful markers for differentiating recent antemortem from postmortem injuries.

Desquamative interstitial pneumonia: an analytic review with an emphasis on aetiology.

Desquamative interstitial pneumonia (DIP) is characterised by the accumulation of numerous pigmented macrophages within most of the distal airspace of the lung and, sometimes, the presence of giant cells. Diagnosis of DIP is not easy and requires surgical lung biopsy. DIP is usually associated with tobacco smoke. However, the association between smoking and DIP is less robust than that with respiratory bronchiolitis with interstitial lung disease or pulmonary Langerhans' cell histiocytosis; approximately 10-42% of patients with DIP are nonsmokers. DIP can also occur in patients following exposure to certain inhaled toxins (occupational exposure) and drugs, and may occur in the context of certain viral illnesses and autoimmune diseases. In the context of DIP, occupational exposure should be systematically investigated.

A combination of smears and cell block preparations provides high diagnostic accuracy for endobronchial ultrasound-guided transbronchial needle aspiration.

Endobronchial ultrasound-guided transbronchial needle aspiration has demonstrated its accuracy in the diagnostic workup of enlarged mediastinal lymph nodes. In addition to conventional smears, the use of liquid-based cytology (LBC) and cell block preparations (CBP) has been introduced more recently. The aim of our study was to determine the performance of each of the different techniques, separately and combined, in terms of diagnostic yield and sensitivity. A total of 290 consecutive patients were included. The pathological examination was based on smear cytology, LBC, and CBP. Adequate sampling was defined by the presence of pathological material or lymphocytes. The global diagnostic yield was 82.7 % and the sensitivity was 89.1 %. The diagnostic yield was 72.8 % for smears, 78.8 % for LBC, and 69.9 % for CBP. The combination of smears with CBP significantly increased diagnostic yield (p = 0.01) and sensitivity (p = 0.006), but not the combination of smears with LBC (yield: p = 0.07; sensitivity: p = 0.13). The combination of the three techniques further increased yield (p = 0.007) and sensitivity (p = 0.006), compared with smears alone. CBP were more sensitive than smears for both diagnoses of carcinoma (p = 0.01) and granulomatous inflammation (p = 0.048). Conversely, LBC was less sensitive than smears for granulomatous inflammation (p = 0.004), but the difference was not significant for carcinoma (p = 0.42). CBP, as a complement to smears, increases diagnostic yield and sensitivity for both diagnoses of carcinoma and granulomatous inflammation. LBC, if used alone, increases the risk of a false-negative result.

BRAF, p53 and SOX2 in anaplastic thyroid carcinoma: evidence for multistep carcinogenesis.

AIMS: The aim of this study was to genotype a series of papillary thyroid carcinomas (PTCs) and anaplastic thyroid carcinomas (ATCs) for BRAF mutation, and to evaluate p53 and SOX2 expression as factors implicated in tumour progression. METHODS: The study included 17 PTCs and 14 ATCs. Analysis of the exon 15 of BRAF was based on direct sequencing. Immunohistochemistry was used to evaluate p53 and SOX2 expression. RESULTS: V600E (c.1799T>A) mutation was observed in 53% (9/17) of PTCs. Two cases of ATCs (2/14; 14%), both with PTC component, harboured BRAF mutation: the classical V600E mutation and an undocumented duplication of codon 599 (c.1795_1797dup; p.Thr599dup). These mutations were present in ATC as well as PTC tumour cells. Overexpression of p53 and SOX2 was depicted respectively in 64% (9/14) and 29% (4/14) of ATCs, and absent in PTCs. CONCLUSION: We confirm that V600E mutation is a frequent and specific event in PTC. BRAF-mutated ATCs are associated with a PTC component displaying the same mutation. We describe a new mutation of BRAF, T599dup, in a case of ATC with tall cell PTC component. Moreover, progression from PTC to ATC could be favoured by further TP53 mutation and SOX2 expression.

[Diagnostic accuracy of endobronchial ultrasound-guided transbronchial needle aspiration in mediastinal lymph nodes]. / Performances diagnostiques de la ponction ganglionnaire médiastinale transbronchique guidée par vidéo écho-endoscopie.

INTRODUCTION: Endobronchial ultrasound-guided transbronchial needle aspiration in mediastinal lymph nodes is a safe method that can be performed for mediastinal and hilar lymph nodes sampling. Because it allows collecting only a small amount of materiel, an optimal processing of the samples is needed. MATERIAL AND METHODS: Based on 150 consecutive procedures, we evaluate the overall diagnostic performances and of each technical methods used for the exploitation of the samples. RESULTS: The global diagnostic yield is 64.0% for the 50 first exams (learning phase), 88.0% for the next 100 exams. The maximal sensitivity and negative predictive value (NPV) are 79.3%, 96.3% and 92.7%, respectively. The yield, sensitivity and NPV of smears are 68.0%, 75.0% and 66.0%, of monolayer preparation 77.8%, 62.1% and 50.0%, of sections from tissue cores of 65.8%, 94.4% and 86.7%. The combination of the different methods increases the yield comparing to tissue cores and smears when taken alone (P < 0.05), and the sensibility and the NPV comparing to smears (P < 0.0005 and P < 0.01, respectively) and monolayer preparation (P < 0.0001 and P < 0.0005, respectively). The sensitivity of tissue cores is greater than smears (P < 0.005) and monolayer preparations (P < 0.0001). This increase in sensitivity is significant for granuloma (sarcoidosis), but not for carcinoma. CONCLUSION: Endobronchial ultrasound-guided transbronchial needle aspiration is an accurate and sensitive technique. Liquid based conditioning of samples and paraffin embedded tissue cores increases the diagnostic performances comparing to smears, notably for the diagnosis of sarcoidosis.