Alice Parisi

Phospho-proteomic analysis of mantle cell lymphoma cells suggests a pro-survival role of B-cell receptor signaling.

BackgroundMantle cell lymphoma (MCL) is currently an incurable entity, and new therapeutic approaches are needed. We have applied a high-throughput phospho-proteomic technique to MCL cell lines to identify activated pathways and we have then validated our data in both cell lines and tumor tissues.MethodsPhosphoScan analysis was performed on MCL cell lines. Results were validated by flow cytometry and western blotting. Functional validation was performed by blocking the most active pathway in MCL cell lines.ResultsPhosphoScan identified more than 300 tyrosine-phosporylated proteins, among which many protein kinases. The most abundant peptides belonged to proteins connected with B-cell receptor (BCR) signaling. Active BCR signaling was demonstrated by flow cytometry in MCL cells and by western blotting in MCL tumor tissues. Blocking BCR signaling by Syk inhibitor piceatannol induced dose/time-dependent apoptosis in MCL cell lines, as well as several modifications in the phosphorylation status of BCR pathway members and a collapse of cyclin D1 protein levels.ConclusionOur data support a pro-survival role of BCR signaling in MCL and suggest that this pathway might be a candidate for therapy. Our findings also suggest that Syk activation patterns might be different in MCL compared to other lymphoma subtypes.

Flat epithelial atypia on core needle biopsy: which is the right management?

The clinical significance and management (surgical excision vs. follow-up) of the patients with the diagnosis of flat epithelial atypia (FEA) on core needle biopsy (CNB) are actually under discussion. Using standardized criteria and precise terminology, we analyzed retrospectively our CNB diagnosis of FEA, dividing patients with pure FEA as the most advanced pathologic lesion from patients with FEA associated to atypical ductal hyperplasia (FEA+ADH). Both the categories were correlated with radiologic data and findings on subsequent surgery. We evaluated 875 core needle biopsies (11-gauge stereotactic vacuum-assisted procedure), performed over a 5-year period. A CNB diagnosis of pure FEA was made in 33/875 (3.7%) cases; in other 11 (1.2%) cases we observed the coexistence of FEA and ADH. Subsequent surgical excisions were available in 20/33 pure FEA and in 10/11 FEA+ADH: of the 20 patients with pure FEA on CNB, none had either ductal carcinoma in situ or invasive carcinoma in their excisional biopsy, whereas 3/10 (30%) FEA+ADH on CNB showed, at subsequent surgery, more advanced lesions (2 ductal carcinoma in situ, 1 invasive carcinoma). Our results suggest that patients with an 11-gauge vacuum-assisted CNB diagnosis of pure FEA (especially if related to a small radiologic target, completely or almost completely removed by the needle biopsy procedure) could be spared surgical excision and managed with close radiologic follow-up.

Short-term outcome after high-intensity focused ultrasound in the treatment of patients with high-risk prostate cancer

To assess the short‐term outcome in patients with high‐risk prostate cancer treated by transrectal high‐intensity focused ultrasound (HIFU).

Signal transduction pathways of mantle cell lymphoma: A phosphoproteome-based study

Mantle cell lymphoma (MCL) is an incurable hematologic malignancy whose pathogenesis is only partly understood. The aim of the present study was to define a “core phosphoproteome” in MCL cell lines that is representative of primary MCL in order to improve knowledge of the signal transduction pathways involved in its tumorigenesis. We have analyzed phosphorylated proteins in several MCL cell lines by immobilized metal affinity chromatography and separation by 2‐D PAGE, followed by RP‐HPLC coupled with MS/MS identification. These data were correlated with information on copy number gains obtained by SNP‐chip analysis. Several of the proteins identified could be linked to a specific signal transduction pathway, and have been recently recognized as important players in MCL pathogenesis, such as nuclear factor‐kappaB (NF‐κB) and phosphoinositide‐3 kinase‐mammalian target of rapamycin (PI3K‐mTOR). However, our data also implicate a number of novel proteins and pathways in the pathobiology of MCL, one of which is mitochondrial signaling. A second‐level analysis identified MAPK1, CK2, CK1, PKCzeta, and PKCepsilon as candidate upstream molecules. Our study provides new insights in MCL pathogenesis and helps to form the basis for testing new target‐specific therapeutics.

BRCA somatic and germline mutation detection in paraffin embedded ovarian cancers by next-generation sequencing

BRCA mutated ovarian cancers respond better to platinum-based therapy and to the recently approved PARP-inhibitors. There is the need for efficient and timely methods to detect both somatic and germline mutations using formalin-fixed paraffin-embedded (FFPE) tissues and commercially available technology. We used a commercial kit exploring all exons and 50bp exon-intron junctions of BRCA1 and BRCA2 genes, and semiconductor next-generation sequencing (NGS) on DNA from 47 FFPE samples of high-grade serous ovarian cancers. Pathogenic mutations were found in 13/47 (28%) cancers: eight in BRCA1 and five in BRCA2. All BRCA1 and two BRCA2 mutations were germline; three BRCA2 mutations were somatic. All mutations were confirmed by Sanger sequencing. To evaluate the performance of the NGS panel, we assessed its capability to detect the 6,953 variants described for BRCA1 and BRCA2 in ClinVar and COSMIC databases using callability analysis. 6,059 (87.1%) variants were identified automatically by the software; 829 (12.0%) required visual verification. The remaining 65 (0.9%) variants were uncallable, and would require 15 Sanger reactions to be resolved. Thus, the sensitivity of the NGS-panel was 99.1%. In conclusion, NGS performed with a commercial kit is highly efficient for detection of germline and somatic mutations in BRCA genes using routine FFPE tissue.

Induction of apoptosis in Jeko-1 mantle cell lymphoma cell line by resveratrol: a proteomic analysis.

Therapies for mantle cell lymphoma (MCL) are clinically unsatisfactory, and the search for effective drugs in vitro might foster the evaluation of their activity in vivo. We have investigated the effects of the polyphenolic compound resveratrol on the MCL cell line Jeko-1 using a combination of flow cytometry, Western blotting and two-dimensional electrophoresis to identify the molecules involved in the induction of apoptosis and cell growth regulation. We show that resveratrol induces apoptosis in Jeko-1 cells and modulates several key molecules, including cyclin D1 (CCND1), p53 (TP53), p21 (CDKN1A), BCL2, BAX, Bcl XL (BCL2L1), caspase 9 (CASP9) and p27 (CDKN1B). By high-resolution 2D-PAGE and nano-reverse phase-high performance liquid chromatography coupled with tandem mass spectrometry, we identified 32 differentially expressed proteins in response to resveratrol treatment that belong to important cell death related networks (including c-myc, NF-kappaB and the mitochondrial apoptotic pathway). These findings may improve the understanding of mechanisms mediating the pro-apoptotic effects of resveratrol on MCL cells, and form the basis for its potential use as a therapeutic agent.

Pulmonary Glomus Tumor

Glomus tumor, also known as glomangioma, is a neoplasm derived from cells of the neuromyoarterial glomus or glomus body. We report a case of glomus tumor of the lung arising in the left lower lobe, incidentally found in a patient who underwent right bilobectomy for a carcinoma localized in the right upper lobe.

Pancreatic granulomatous necrotizing vasculitis: a case report and review of the literature

This paper describes a 48-year-old woman admitted to surgical department with midepigastric pain. Ultrasonography of the abdomen and magnetic resonance imaging revealed an ipoechoic mass in the head of pancreas suggestive for malignant tumor. She underwent pancreaticoduodenectomy; histologic examination showed a granulomatous inflammation with necrosis and destruction of the wall of small-medium size blood vessels very suggestive for Wegener’s granulomatosis (WG). No other visceral involvement was found; test for antineutrophil cytoplasmic antibodies was negative. Initial and symptomatic involvement of pancreas was reported in very few cases of WG.

Endoscopic ultrasound features of pancreatic schwannoma.

A 59-year-old female underwent abdominal ultrasound for dyspepsia. A 2 cm pancreatic nodule was incidentally discovered. Computer tomography scanning confirmed a solid mass of the pancreatic uncinate process; at endoscopic ultrasound (EUS) examination, it appeared as a round, well-demarked, solid homogeneous, hypoechoic mass [Figure 1]. In contrast-enhanced EUS (CE-EUS) images, the lesion showed poor contrast intake, with a hypoenhanced pattern compared to the surrounding pancreatic parenchyma [Figure 2]. Fine-needle aspiration (FNA) with a standard 25G needle revealed spindle cells [Figure 3] that expressed S-100 protein, suggestive for schwannoma. Surgical enucleation was performed, and diagnosis was confirmed on a surgical specimen [Figure 4].

Pancreatic Lesions: Pathologic Correlations

In recent years the evolution of imaging technology has made possible a more detailed identification of the structural features of pancreatic cancers, facilitating the differential diagnosis between different tumor histologies and pancreatic diseases in relation to other tumor-forming lesions. At the same time pathologists have provided a most accurate definition of the different tumor histologies with the development of the most recent WHO classification (2010) [1].