High-throughput somatic mutation profiling in pulmonary sarcomatoid carcinomas using the LungCarta™ Panel: exploring therapeutic targets.

BACKGROUND: Pulmonary sarcomatoid carcinomas (SC) are tumors characterized by poor prognosis and resistance to conventional platinum-based chemotherapy. This study sought to describe the mutational profile of SC using high-throughput genotyping technology. PATIENTS AND METHODS: We used mass spectrometry to test 114 surgical biopsies from 81 patients with SC for 214 mutations affecting 26 oncogenes and tumor suppressor genes. RESULTS: In total, 75 (92.6%) patients were smokers. Within the total 81 tumors, 67 distinct somatic alterations were identified, with 56 tumors (69.1%) harboring at least one mutation. The most frequent mutations were KRAS (27.2%), EGFR (22.2%), TP53 (22.2%), STK11 (7.4%), NOTCH1 (4.9%), NRAS (4.9%), and PI3KCA (4.9%). The EGFR mutations were almost always rare mutations (89%). In 32 tumors (39.5%), two or more mutations co-existed, with up to four mutations in a single case. In six different cases, comparative genetic analysis of different histological areas from the same tumor (giant, spindle, or epithelial component) revealed a 61% concordance rate for all the mutations with a 10% detection threshold, compared with 91.7% with a 20% detection threshold. CONCLUSION: Our results demonstrated a high mutation rate and frequent co-mutations. Despite SC tumors exhibiting a high histological heterogeneity, some intratumoral molecular homogeneity was found. Now with newly developed targeted therapies, SC patients may be eligible for new target mutations, and can now therefore be screened for clinical trials.

[The French mesothelioma network from 1998 to 2013]. / Mésothéliome: les dispositifs en place en France « le réseau mésothéliome ¼ 1998-2013.

Mesothelioma is a rare disease less than 0.3% of cancers in France, very aggressive and resistant to the majority of conventional therapies. Asbestos exposure is nearly the only recognized cause of mesothelioma in men observed in 80% of case. In 1990, the projections based on mortality predicted a raise of incidence in mesothelioma for the next three decades. Nowadays, the diagnosis of this cancer is based on pathology, but the histological presentation frequently heterogeneous, is responsible for numerous pitfalls and major problems of early detection toward effective therapy. Facing such a diagnostic, epidemiological and medico-legal context, a national and international multidisciplinary network has been progressively set up in order to answer to epidemiological survey, translational or academic research questions. Moreover, in response to the action of the French Cancer Program (action 23.1) a network of pathologists was organized for expert pathological second opinion using a standardized procedure of certification for mesothelioma diagnosis. We describe the network organization and show the results during this last 15years period of time from 1998-2013. These results show the major impact on patient's management, and confirm the interest of this second opinion to provide accuracy of epidemiological data, quality of medico-legal acknowledgement and accuracy of clinical diagnostic for the benefit of patients. We also show the impact of these collaborative efforts for creating a high quality clinicobiological, epidemiological and therapeutic data collection for improvement of the knowledge of this dramatic disease.

Identification and characterization of two genes (MIP-1beta, VE-CADHERIN) implicated in acute rejection in human heart transplantation: use of murine models in tandem with cDNA arrays.

BACKGROUND: Genes and mechanisms of action involved in human acute rejection after allogeneic heart transplantation remain to be elucidated. The use of a murine allograft model in tandem with cDNA arrays and quantitative real-time polymerase chain reaction (Q-PCR) can greatly help in identifying key genes implicated in human heart acute rejection. METHODS AND RESULTS: Hearts from Balb/c mice were either not transplanted or transplanted heterotopically in the abdomen of Balb/c (isografts) and C57BL/6 (allografts) mice. Histological analysis showed acute rejection only in allografts. Total RNA was extracted from isografts (n=3), allografts (n=4), and not transplanted hearts (n=4); reverse transcribed; and labeled with P32. Each probe was hybridized to cDNA macroarrays. Eight genes were overexpressed and 7 genes were underexpressed in allografts compared with isografts. Macrophage inflammatory protein-1beta (MIP-1beta), an overexpressed gene, and VE-cadherin, an underexpressed gene, were validated by immunohistochemistry and Q-PCR in the murine models. Genes of interest, validated in the 3 murine groups, were then investigated in human heart tissues. Immunohistochemistry and Q-PCR performed on endomyocardial biopsies after heart transplantation showing no rejection (n=10) or grade IB (n=10) or IIIA (n=10) rejection, according to International Society of Heart and Lung Transplantation criteria, confirmed the results obtained from the murine model. CONCLUSIONS: We have demonstrated that the upregulation of MIP-1beta and downregulation of VE-cadherin may strongly participate in human acute heart rejection.

Polishing surgical metal pieces, granulomatosis and mineralogical analysis.

This report describes the case of a 44-year-old man with pulmonary nodules whose histological analysis initially suggested tuberculosis. The Mycobacterium tuberculosis (MT) culture was negative and a questionnaire revealed a professional activity of brushing and polishing surgical instruments without any protection for 7 years.  A mineralogical analysis by optical and electron microscopy was performed on both a healthy lung tissue biopsy and a lung nodule in a paraffin block. Electron microscopy analysis revealed the presence of metal particles (iron oxide, titanium oxide, aluminum oxide and steel) in both samples. This study suggests that mineralogical analysis combined with a questionnaire on dust exposure could help redirect the diagnosis of a dust-related disease.

[Pulmonary lymphangioleiomyomatosis: From pathogenesis to management]. / Lymphangioléiomyomatose pulmonaire : de la physiopathologie à la prise en charge.

INTRODUCTION: Pulmonary lymphangioleiomyomatosis (LAM) is a rare disease affecting mainly young women. BACKGROUND: The respiratory manifestations are characterized by a progressive cystic destruction of the lung parenchyma. Extrapulmonary involvement includes benign renal tumours called angiomyolipomas and abdominal lymphatic masses called lymphangioleiomyomas. At the pathological level, the cellular proliferation found in LAM is in part due to the presence of mutations in the tumour suppressor genes TSC1 and TSC2 (Tuberous Sclerosis Complex). These mutations lead to the activation of the mTOR pathway, which is currently the main therapeutic target. mTOR inhibitors such as sirolimus or everolimus have shown a beneficial effect on the decline in pulmonary function and a reduction of angiomyolipoma size, but are necessary in only some patients. PERSPECTIVES: LAM cells have migratory properties mediated by the formation of new lymphatic vessels. They are also able to secrete metalloproteases, which enhance their invasiveness. Moreover, the expression of estrogen and progesterone receptors by LAM cells suggests a possible role for sex hormones in the pathogenesis of the disease. CONCLUSION: A better understanding of mTOR-independent mechanisms would allow the development of novel therapeutic approaches.

[Elaboration of a national biobank for the study of gestational trophoblastic diseases]. / Création d'une biobanque nationale pour l'étude des maladies trophoblastiques gestationnelles.

AIM: To generate a national biobank made up of samples of the highest quality for the purpose of inciting basic research on gestational trophoblastic diseases (GTD). MATERIAL AND METHODS: Three priority axes of research were defined to optimize the nature, method of collection, and storage of the samples. These are: to enhance our understanding of GTD, develop new diagnostic tests, and identify new therapeutic targets. The protocol for patient inclusion and sample processing was determined after extensive literature review and collaboration with international experts in the field of GTD. RESULTS: For each patient with a GTD and for control patients (legally induced abortions), chorionic villi, decidua and tumor samples (fresh, immersed in RNA-protective solution and fixed in formaldehyde), blood (serum, plasma, RNA, and peripheral blood mononuclear cells), urine (supernatant), and cell cultures of villous cytotrophoblasts are prospectively collected. Associations are then made between the collected samples and numerous clinical and biological data, such as human chorionic gonadotropic plasma levels following curettage in the case of a hydatidiform mole. CONCLUSION: Such a collection of high quality samples and their associated data open up new perspectives for both national and international collaborative research projects.

Value of ultrasound-guided fine-needle aspiration in the management of ovarian and paraovarian cysts.

This study was designed to assess if cytology was accurate for an appropriate diagnosis of ovarian and paraovarian cysts, and if the ultrasound-cytology-estradiol (UCE) triad was sufficient to discriminate functional vs. nonfunctional cysts, the latter requiring surgical resection. One hundred twenty-two ultrasound-diagnosed adnexal cysts were punctured and surgically removed, and then subjected to cytologic and histologic examinations; 90 of these fluids were assayed for estradiol. Histologically, 30 cysts were functional and 92 were nonfunctional. A correct discrimination between functional and nonfunctional origin was obtained in 54.9% of cases with cytology, in 94.4% with estradiol assay, in 50.8% with ultrasonography, and in 97.8% with these three examinations combined (UCE triad). Among the 34 patients with no criteria of neoplastic origin (age >40, ultrasonographic findings), the UCE triad diagnosed six functional cysts. Therefore, 17.6% (6/34) of these young women could have avoided unnecessary surgery. Diagn. Cytopathol. 2000;22:70-80.

Cytological aspiration of intraocular retinoblastoma in an 11-year-old boy.

Only a few cases of retinoblastomas in older children have been reported and the clinical diagnosis may be difficult. In case, fine-needle aspiration from an atypical retinal mass of an 11-yr-old boy was performed. The vitreous fluid was stained with Diff-Quik for an immediate cytological examination and the diagnosis of retinoblastoma was suggested. The rest of the specimen was separated into two parts. One was stained with May-Grünwald-Giemsa and the other was centrifuged, embedded in paraffin, and finally stained with hematoxylin-eosin-safran. The undifferentiated blue cells were associated with abundant necrotic debris and portions of capillaries with perivascular tumor cells around. The cytoplasm of the tumor cells was strongly stained with neuron-specific enolase antibody. The diagnosis of retinoblastoma was confirmed. The specimen of enucleation confirmed the diagnosis. In conclusion, cytological aspiration can categorically diagnose suspected intraocular tumors of older children in whom clinical and noninvasive investigations have failed to establish the diagnosis.

Fine needle aspiration cytology of a solid papillary carcinoma of the breast. A case report with immunohistochemical studies.

BACKGROUND: Solid papillary carcinoma of the breast was recently described. This tumor display distinctive clinical and morphologic features. It is an intraductal papillary carcinoma frequently associated with mucinous carcinoma and infiltrating ductal carcinoma not otherwise specified. CASE: Fine needle aspiration cytology (FNAC) of a solid papillary carcinoma of the right breast in an 82-year-old female, demonstrated a cellular specimen of discohesive, small, uniform and ovoid tumor cells, with occasional loose cluster. The cytoplasm was abundant and finely granular. The nuclei were round, without marked atypia. The nucleoli were inconspicuous. SCant mucinous material was present. The diagnosis was confirmed histologically and immunohistochemically. CONCLUSION: The cytologic aspects of solid papillary carcinoma of the breast suggest a carcinoma with endocrine differentiation and a weak mucinous component.

[Primary pulmonary synovial sarcoma. Apropos of 1 case with cytogenetic study]. / Synovialosarcome pulmonaire primitif. A propos d'un cas avec étude cytogénétique.

Synovial sarcoma most commonly occurs in the peri-articular regions of the extremities. We report a case of primary pulmonary monophasic synovial sarcoma. This tumor is extremely rare and shows a particular immunohistochemical pattern of great help for the diagnosis. Cytogenetic study confirm the diagnosis by showing the specific t (X; 18) chromosomal translocation, characteristic of synovial sarcoma in all anatomic locations.

[Malignant mesothelioma with osteoblastic heterologous elements]. / Mésothéliome malin avec éléments hétérologues ostéoblastiques.

A 59 year-old man with a history of asbestos exposure presented with a right pleural effusion and a diffuse pleural thickening with focal calcifications on chest X-ray. Cytological examination of pleural fluid indicated malignant mesothelioma. A biopsy specimen showed malignant mesothelioma surrounding a fragment of mature bone. The patient was treated with intrapleural interferon, but relapsed 3 years later. A fresh biopsy specimen showed round tumor cells surrounding osteoid substance. Only ten cases of this rare variant of malignant mesothelioma with osteoblastic heterologous elements have been reported in the literature. The most difficult differential diagnosis is primary pleural osteosarcoma.

[Glomangioma: an unusual ethmoid-nasal tumor]. / Glomangiome: une tumeur ethmoïdo-nasale inhabituelle.

Glomus tumors are benign vascular tumors usually located in the fingertip dermis. Extracutaneous glomus tumors are extremely rare. A new case of sino-nasal glomangioma is reported and both clinical and histological features and biological behavior of glomus tumors in this exceptional site are described.

[Transthoracic aspiration. Evaluation of cytologic and histologic diagnosis in a pulmonary nodule by retrospective comparison of 2 series of 267 cytoaspirations and 292 coaxial needle aspirations]. / La ponction trans-thoracique. Evaluation du diagnostic cytologique et histologique d'un nodule pulmonaire par comparaison rétrospective de 2 séries de 267 cytoponctions et de 292 ponctions à l'aiguille coaxiale.

Percutaneous fine-needle aspiration is a well established method for the diagnosis of peripheral lung lesion. In order to compare different methods of aspiration, we analyze retrospectively two different series: 267 fine needle aspirations (FNA) compared with the histological diagnosis on surgical specimens and 292 lung biopsies using a coaxial technique with comparison between cytological diagnosis--smears and imprints--and histological diagnosis simultaneously obtained on the same specimen. The sensitivity (91%), the specificity (90%) and the overall typing accuracy related to the histological types obtained by FNAB are equivalent to those of the literature. The low rate of pneumothorax in the series (6%) is related to the use of immediate interpretation of the specimen. Automated biopsy with a coaxial cutting needle provide cytological specimens--smears and imprint--with a high rate of sensibility (95.3%) and of sensibility (98%). The overall sensitivity of the cytological methods alone is better than biopsy (95.3% vs. 92.9%), but the typing accuracy is not as good as biopsy alone (98% vs. 100%). False-positive and false-negative diagnoses are the same in both series. In conclusion the percutaneous aspiration method choose to establish a morphological diagnosis in lung lesion depends now on the habits of the radiologist and the pathologist.

[Histopathologic study of melanoma of the choroid after proton therapy]. / Etude histopathologique des mélanomes choroïdens après protonthérapie.

To evaluate irradiation effects on choroidal melanomas, histopathologic findings of 18 eyes whose primary treatment was enucleation were compared to 15 eyes enucleated after proton beam irradiation. Irradiated tumors showed more likely necrosis (p = 0.01) had balloon cells (p = 0.01), and inflammatory infiltrate (p = 0.05). In the irradiated group, the prevalence of tumor blood vessel damage was higher (p = 0.0002) and mitotic figures were fewer (p = 0.01). These findings suggest that proton beam irradiation damages tumor cells and alters the tumor's capacity for cellular reproduction. It damages blood vessels leading to tumor necrosis. It induces an inflammatory response of unknown effects. Radiosensitivity of choroidal melanomas cannot be assessed using conventional histologic methods. However, tumor necrosis, mitotic activity and rate of balloon cells can help to establish tumoral sensitivity to irradiation.

[Sarcoma of the pulmonary artery]. / Sarcomes de l'artére pulmonaire.

Two cases of pulmonary artery angiosarcomas are reported. Pulmonary artery tumor was diagnosed during exploratory thoracotomy in the first case. It was suspected on magnetic resonance imaging and confirmed by surgery in the second case. The therapeutic was a right pneumonectomy for the first patient and a tumor resection followed by chemotherapy and radiotherapy on cerebral metastases for the second patient. Both our patients died, 13 and 44 months after surgery. On the basis of histological and immunohistochemical findings both tumors were angiosarcomas. Ultrastructural examination in the second case revealed an endothelial differentiation. The review of 142 previously reported cases of pulmonary artery sarcomas and these ultrastructural findings suggest that pulmonary artery sarcomas may have multiple origins; leiomyosarcomas deriving from arterial smooth muscle; angiosarcomas deriving from the endothelium and undifferentiated sarcomas or "myointimal sarcomas" deriving from myointimal cells.

[Bronchiolo-alveolar carcinoma and lung transplantation]. / Carcinome bronchioloalvéolaire et transplantation pulmonaire.

INTRODUCTION: Bronchiolo-alveolar carcinoma is a controversial indication for lung transplantation because of the risk of recurrence. We report three cases and propose some risk factors for recurrence. CASE REPORTS: Our study concerns three patients transplanted at the Louis-Pradel Hospital between 1991 and 2010. The first patient relapsed 86 months after transplantation, benefited from surgical treatment, then died of renal failure. A second patient died of infection, without recurrence, 72 months after transplantation. The third had an early recurrence at 7 months and died 27 months after transplantation. The risk factors for recurrence appear to be: clinically "aggressive" presentation and histological stromal pulmonary invasion by the carcinoma. CONCLUSION: Diffuse bronchiolo-alveolar carcinoma is a possible indication of lung transplantation. The risk of recurrence imposes some requirements: a precise histological diagnosis and a slow clinical course.