Duplications of KIAA1549 and BRAF screening by Droplet Digital PCR from formalin-fixed paraffin-embedded DNA is an accurate alternative for KIAA1549-BRAF fusion detection in pilocytic astrocytomas

Pilocytic astrocytomas represent the most common glioma subtype in young patients and account for 5.4% of all gliomas. They are characterized by alterations in the RAS–MAP kinase pathway, the most frequent being a tandem duplication on chromosome 7q34 involving the BRAF gene, resulting in oncogenic BRAF fusion proteins. BRAF fusion involving the KIAA1549 gene is a hallmark of pilocytic astrocytoma, but it has also been recorded in rare cases of gangliogliomas, 1p/19q co-deleted oligodendroglial tumors, and it is also a common feature of disseminated oligodendroglial-like leptomeningeal neoplasm. In some difficult cases, evidence for KIAA1549-BRAF fusion is of utmost importance for the diagnosis. Moreover, because the KIAA1549-BRAF fusion constitutively activates the MAP kinase pathway, it represents a target for drugs such as MEK inhibitors, and therefore, the detection of this genetic abnormality is highly relevant in the context of clinical trials applying such new approaches. In the present study, we aimed to use the high sensitivity of Droplet Digital PCR (DDPCR™) to predict KIAA1549-BRAF fusion on very small amounts of formalin-fixed paraffin-embedded tissue in routine practice. Therefore, we analyzed a training cohort of 55 pilocytic astrocytomas in which the KIAA1549-BRAF fusion status was known by RNA sequencing used as our gold standard technique. Then, we analyzed a prospective cohort of 40 pilocytic astrocytomas, 27 neuroepithelial tumors remaining difficult to classify (pilocytic astrocytoma versus ganglioglioma or diffuse glioma), 15 dysembryoplastic neuroepithelial tumors, and 18 gangliogliomas. We could demonstrate the usefulness and high accuracy (100% sensitivity and specificity when compared to RNA sequencing) of DDPCR™ to assess the KIAA1549-BRAF fusion from very low amounts of DNA isolated from formalin-fixed paraffin-embedded specimens. BRAF duplication is both necessary and sufficient to predict this fusion in most cases and we propose that this single analysis could be used in routine practice to save time, money, and precious tissue

Negative Survival Impact of High Radiation Doses to Neural Stem Cells Niches in an IDH-Wild-Type Glioblastoma Population

Aims: Assess the impact of radiation doses to neural stem cell (NSC) niches in patients with IDH-wild-type glioblastoma.Materials and Methods: Fifty patients were included in the study. NSC niches [SubVentricular Zone (SVZ) and Sub Granular Zone (SGZ)] were contoured by fusing CT scans and pre-therapy MRI, Tumor location defined ipsilateral and contralateral SVZ and SGZ. Prognostic significance of clinical, biological and dosimetric parameters were examined. We generated a Recursive Partitioning Analysis (RPA) model with independent prognostic classes.Results: Median follow-up: 23.8 months. Event free and overall survival (OS): 10 and 19.1 months. Incomplete surgery, PTV (planning target volume), ipsilateral SVZ or NSC niche mean dose > 57.4 Gy, contralateral NSC niche mean dose > 35 Gy and bilateral NSC niche mean dose > 44 Gy were significantly correlated with reduced OS. Only EGFR amplification was an independent prognostic factor (p = 0.019) for OS. RPA generated independent risk groups: 1 (low risk): [ipsilateral NSC mean dose (INMD) < 58.01 Gy and methylated MGMT promoter], 2: (INMD < 58.01 Gy and unmethylated MGMT promoter and contralateral SVZ mean dose < 18.6 Gy; p = 0.43), 3: (INMD < 58.01 Gy and unmethylated MGMT promoter and contralateral SVZ mean dose > 18.6 Gy; p = 0.002) and 4: (very high risk) (INMD > 58.01 Gy; p < 0.001).Conclusion: High radiation doses to ipsilateral NSC and contralateral SVZ could have a negative impact on overall survival in IDH-wild-type glioblastoma population

AIRE-Deficient Patients Harbor Unique High-Affinity Disease-Ameliorating Autoantibodies

APS1/APECED patients are defined by defects in the autoimmune regulator (AIRE) that mediates central T cell tolerance to many self-antigens. AIRE deficiency also affects B cell tolerance, but this is incompletely understood. Here we show that most APS1/APECED patients displayed B cell autoreactivity toward unique sets of approximately 100 self-proteins. Thereby, autoantibodies from 81 patients collectively detected many thousands of human proteins. The loss of B cell tolerance seemingly occurred during antibody affinity maturation, an obligatorily T cell-dependent step. Consistent with this, many APS1/APECED patients harbored extremely high-affinity, neutralizing autoantibodies, particularly against specific cytokines. Such antibodies were biologically active in vitro and in vivo, and those neutralizing type I interferons (IFNs) showed a striking inverse correlation with type I diabetes, not shown by other anti-cytokine antibodies. Thus, naturally occurring human autoantibodies may actively limit disease and be of therapeutic utility.Peer reviewe

Biology of mesenchymal tumors of intermediate malignancy : the meningeal solitary fibrous tumor model

Le concept de tumeur conjonctive de malignité intermédiaire a été introduit par les classifications proposées par l’organisation mondiale de la santé (OMS): la tumeur fibreuse solitaire, une néoplasie conjonctive rare, est un modèle intéressant pour cette problématique. L’objectif général de ces travaux était de mieux définir la biologie de ces tumeurs par une approche multimodale, intégrant histopathologie, immunohistochimie et biologie moléculaire afin d'affiner leur pronostic et diagnostic.D’un point de vue diagnostique, nos travaux ont confirmé les excellentes performances de l’immunohistochimie STAT6 et sa sensibilité supérieure par rapport à la RT-PCR ciblée. D’un point de vue pronostique, nous avons précisé les paramètres cliniques, histologiques et moléculaires qui impactent la survie des patients : l’état des limites d’exérèse, l’index mitotique et la nécrose sont déterminants, d’autant plus s’ils sont combinés. Ainsi, nous avons montré qu’une révision du grade MGS, basée sur une combinaison facteurs histologiques permettaient d’affiner le pronostic des formes méningées. Nos résultats suggèrent que l’ensemble du spectre est de malignité intermédiaire, dès lors que la durée de suivi est suffisante. Nos travaux ont aussi montré que le type de transcrit de fusion NAB2-STAT6 et les mutations du promoteur de TERT ne présentent pas de valeur pronostique.Ainsi, le diagnostic et le pronostic des tumeurs fibreuses solitaires méningées peut se baser sur des critères histologiques accessibles bien que leur évaluation pronostique ne soit pas encore complètement résolue.Mesenchymal neoplasms of intermediate malignancy were introduced by the World Health Or-ganization (WHO) classification of soft-tissue and bone tumors. This terminology accounts for a heterogeneous group of neoplasms, which common denominator is to fail usual prognostic approaches. Solitary fibrous tumor, a rare neoplasia, is an interesting model of such tumors with a potential for local recurrence and a low risk of metastasis. In this context, the main goal of this work was to define the biology of these neoplasms, using a multimodal approach. Considering their diagnosis, our work confirmed the excellent performance of STAT6 immunohistochemistry and its greater sensitivity compared to targeted RT-PCR. Considering the prognosis, we further specified the clinical, histological and molecular parameters that impact survival: surgical margin, mitotic index and necrosis. We revised the criteria of the Marseille Grading Scheme (MGS) accordingly, which resulted in high prognosis value for meningeal forms. Our results point that the whole spectrum is of intermediate malignancy as long as the follow-up is sufficient to allow detection of delayed adverse events. Our works also showed that the type of NAB2-STAT6 tran-script and the mutation of TERT promoter do not carry an independent prognostic value, albeit certain types of transcripts were associated with particular clinical and histological criteria. To conclude, the diagnosis and prognosis assessment of solitary fibrous tumors can be based on accessible histological criteria albeit their prognosis assessment is not fully satisfactory yet

Cutaneous Melanomas Arising during Childhood: An Overview of the Main Entities

International audienceCutaneous melanomas are exceptional in children and represent a variety of clinical situations, each with a different prognosis. In congenital nevi, the risk of transformation is correlated with the size of the nevus. The most frequent type is lateral transformation, extremely rare before puberty, reminiscent of a superficial spreading melanoma (SSM) ex-nevus. Deep nodular transformation is much rarer, can occur before puberty, and must be distinguished from benign proliferative nodules. Superficial spreading melanoma can also arise within small nevi, which were not visible at birth, usually after puberty, and can reveal a cancer predisposition syndrome (CDKN2A or CDK4 germline mutations). Prognosis is correlated with classical histoprognostic features (mainly Breslow thickness). Spitz tumors are frequent in adolescents and encompass benign (Spitz nevus), intermediate (atypical Spitz tumor), and malignant forms (malignant Spitz tumor). The whole spectrum is characterized by specific morphology with spindled and epithelioid cells, genetic features, and an overall favorable outcome even if a regional lymph node is involved. Nevoid melanomas are rare and difficult to diagnose clinically and histologically. They can arise in late adolescence. Their prognosis is currently not very well ascertained. A small group of melanomas remains unclassified after histological and molecular assessment

Acral FibroChondroMyxoid tumor: imaging features of a new entity

International audience"Acral FibroChondroMyxoid tumor" (AFCMT) is a recently described distinctive subtype of acral soft tissue tumor that typically arises on the fingers and toes. We herein present the unreported imaging features of AFCMT in a 44-year-old woman. This otherwise healthy patient was referred for a painful, slow-growing, soft tissue mass in the middle finger of her right hand. Initial radiographs and computed tomography showed a small lesion centered in the soft tissue of the ulnar aspect of the proximal phalanx, associated with scalloping of the underlying bone. Magnetic resonance imaging confirmed the presence of a well-circumscribed soft tissue tumor that exhibited relatively high T2-weighted signal intensity and marked enhancement after contrast administration. Subsequent excisional biopsy was performed. Histologically, the tumor was characterized by an abundant stroma displaying fibrous, chondroid, and myxoid areas. By immunohistochemistry, tumor cells stained for CD34, ERG, and focally S100 protein. RNA-sequencing allowed detection of THBS1-ADGFR5 gene fusion which confirmed the diagnosis of AFCMT. At 2-year follow-up, the patient remains free of recurrence. AFCMT is a previously unrecognized entity that may mimic chondroma and should be considered in the differential diagnosis of soft tissue tumors with cartilaginous or myxoid stroma in the extremities

Point de vue : une leçon tirée du confinement, l’histopathologie en visioconférence

International audienceThe use of videoconferencing had increased significantly during lockdown. During this period, videoconferencing has been used in the pathological department of pathology (Timone university hospital, Marseille, France) for academic, diagnosis and referral. We provide our point of view regarding the use of this tool. As discussing slides through videoconferencing is a new and specific activity, we have also summarised specific recommendations for practical remote histopathology meetings.Le confinement et la distanciation sociale ont généralisé l’usage de la visioconférence, tant dans la sphère privée que professionnelle. Ainsi, l’utilisation de la visioconférence a considérablement augmenté durant la période du confinement et cette technologie a été utilisée dans le service d’anatomie pathologique du centre hospitalo-universitaire Timone, à Marseille, à visée universitaire, hospitalière et de recours. Nous apportons notre point de vue concernant l’utilisation de cet outil informatique. La discussion de lames par visioconférence est un exercice nouveau et particulier ; plusieurs recommandations sont émises pour un bon déroulement de ces réunions d’histopathologie à distance

De multiples nodules abdominaux calcifiés chez une femme

International audienc