Riccardo Valli

Reproducibility of the WHO histological criteria for the diagnosis of Philadelphia chromosome-negative myeloproliferative neoplasms

This study, performed on behalf of the Italian Registry of Thrombocythaemias (Registro Italiano Trombocitemie), aimed to test the inter-observer reproducibility of the histological parameters proposed by the WHO classification for the diagnosis of the Philadelphia chromosome-negative myeloproliferative neoplasms. A series of 103 bone marrow biopsy samples of Philadelphia chromosome-negative myeloproliferative neoplasms consecutively collected in 2004 were classified according to the WHO criteria as follows: essential thrombocythaemia (n=34), primary myelofibrosis (n=44) and polycythaemia vera (n=25). Two independent groups of pathologists reviewed the bone marrow biopsies. The first group was asked to reach a collegial ‘consensus’ diagnosis. The second group reviewed individually all the cases to recognize the main morphological parameters indicated by the WHO classification and report their results in a database. They were subsequently instructed to individually build a ‘personal’ diagnosis of myeloproliferative neoplasms subtype just assembling the parameters collected in the database. Our results indicate that high levels of agreement (≥70%) have been reached for about all of the morphological features. Moreover, among the 18 evaluated histological features, 11 resulted statistically more useful for the differential diagnosis among the different Philadelphia chromosome-negative myeloproliferative neoplasms. Finally, we found a high percentage of agreement (76%) between the ‘personal’ and ‘consensus’ diagnosis (Cohen’s kappa statistic >0.40). In conclusion, our results support the use of the histological criteria proposed by the WHO classification for the Philadelphia chromosome-negative myeloproliferative neoplasms to ensure a more precise and early diagnosis for these patients.

Methylation changes of SIRT1, KLF4, DAPK1 and SPG20 in B-lymphocytes derived from follicular and diffuse large B-cell lymphoma

Diffuse large-B cell lymphomas (DLBCL) and follicular lymphomas (FL) are the most represented subtypes among mature B-cell neoplasms and originate from malignant B lymphocytes. Methylation represents one of the major epigenetic mechanisms of gene regulation. Silent information regulator 1 (SIRT1) is a class III lysine-deacetylase playing several functions and considered to be a context-dependent tumor promoter. We present the quantitative methylation, gene expression and tissue distribution of SIRT1 and some key mediators related to lymphoma pathogenesis in B lymphocytes purified from biopsies of follicular hyperplasias, FL and DLBCL. SIRT1 mRNA levels are higher in FL than follicular hyperplasias and DLBCL. B cell lymphoma 6 (BCL6) positively correlates with SIRT1. SIRT1 promoter shows a methylation decrease in the order: follicular hyperplasia - FL - DLBCL. Kruppel-like factor 4 (KLF4), Death-associated protein kinase 1 (DAPK1) and Spastic Paraplegia 20 (SPG20) methylation increase significantly in FL and DLBCL compared to follicular hyperplasias. Gene expression of DAPK1 and SPG20 inversely correlates with their degree of methylation. Our findings evidence a positive correlation between SIRT1 and BCL6 expression increase in FL. SIRT1 methylation decreases in FL and DLBCL accordingly and this parallels the increase of KLF4, DAPK1 and SPG20 methylation.

Anaplastic Lymphoma Kinase–Positive Large B-Cell Lymphoma Description of a Case With an Unexpected Clinical Outcome

Anaplastic lymphoma kinase–positive (ALK-positive) large B-cell lymphoma is a rare and aggressive variant of large B-cell lymphoma (LBCL), first reported by Delsol et al in 1997, showing distinctive morphologic, immunophenotypic and cytogenetic features. The latest 2008 World Health Organization Classification of Tumours of Haematopoietic and Lymphoid tissues recognizes ALK-positive LBCL as a separate entity. Here, we report a case of ALK-positive large B-cell lymphoma in a 53-year-old man with diffuse abdominal and mediastinal lymph-nodes involvement. According to the Ann Arbor staging system, the patient had a stage IIIB lymphoma. The age-adjusted International Prognostic Index was 2 (stage III and elevated lactate dehydrogenase), so the disease was considered high risk. The patient underwent chemotherapy, radiotherapy, and an autologous stem cell transplantation. The patient is alive and free of disease 35 months after diagnosis.

A unique case of an indolent myometrial T-Cell lymphoproliferative disorder with phenotypic features resembling uterine CD8+ resident memory T cells

Objective: Indolent extranodal T-cell lymphoproliferative disorders have recently been described as new entities in the gastrointestinal tract and acral sites displaying clonal T-cell receptor (TCR) rearrangement and nonactivated cytotoxic CD8+ T-cell phenotypes. Methods/Results: We report a unique case of an atypical myometrial T-cell lymphoproliferation in a 39-year-old multiparous woman, which shares many of the features mentioned above: CD8+/TIA1+/granzyme B- phenotype, clonal TCR rearrangement and indolent course. Conclusion(s): We hypothesize that it might derive from a subset of uterine nonrecirculating CD8+ resident memory T cells expanded after repeated exposure to allo-extravillous trophoblastic antigen.

Comparison of JAK2V617F-positive essential thrombocythaemia and early primary myelofibrosis: The impact of mutation burden and histology

An accurate histological diagnosis may distinguish essential thrombocythaemia (ET) from early primary myelofibrosis (early‐PMF), which is associated with worse outcome. Outcome of ET is also negatively affected by the presence of the JAK2V617F mutation. To investigate the impact of JAK2V617F mutation burden and histology on outcome, we collected 475 WHO‐diagnosed ET (69.2%) or early‐PMF JAK2V617F‐positive patients followed in 4 Italian haematology centers. JAK2V617F allele burden was ≤50% in 90% and 87% of ET and early‐PMF patients, respectively (P = .34). During follow‐up, 32 (9.7%) ET and 18 (12.3%) early‐PMF patients experienced 59 thrombotic events, and 27 patients (5.6%) and 6 (1.2%) patients evolved to myelofibrosis and acute leukemia, respectively. At last contact, 28 (5.8%) patients had died. In early‐PMF compared to ET, the 10‐year mortality rates (6.7% and 4.3%, P = .73), leukemic transformation rates (1.4% and 1.2%, P = .45), and thrombosis rates (16.7% and 12.2%, P = .12) were comparable. Only progression to overt myelofibrosis at 10 years was significantly worse (11.4% and 1.5%, P = .004). In multivariate analysis, a higher (>50%) JAK2V617F burden was significantly correlated with fibrotic progression and histology. Considering JAK2V617F‐positive disease, a higher (>50%) JAK2V617F burden and histological classification are independent prognostic risk factors for disease progression. These findings reinforce the need for standardized detection of this mutation.

Abstract 4451: Different patterns of SIRT1, KLF4, DAPK1 and SPG20 methylation in B lymphocytes correlate with the clinical parameters of non-Hodgkin lymphoma subtypes

Background. DNA methylation is one of the best studied epigenetic modifications and one major constituent of the epigenome of a cell. It contributes to normal development as well to carcinogenesis. Nowadays, many efforts are being made in order to use DNA methylation as a biomarker. The aim of our work is to characterize the expression and methylation of SIRT1, HIC1, BCL6, KLF4 and other genes relevant for Non-Hodgkin lymphomas (NHL) pathogenesis. Methods. Immunohistochemistry (IHC) on 72 formalin-fixed paraffin embedded tissue sections (FFPE). B-lymphocytes were purified from 36 biopsies of follicular hyperplasias (non-malignant B-lymphocytes), follicular lymphomas (FL) and diffuse large B-cell lymphomas (DLBCL). Gene expression were analysed by quantitative retrotranscribedPCR (qRTPCR). Quantitative CpG promoter methylation analysis was performed by pyrosequencing after bisulfite conversion or by Methyl II array qPCR on genomic DNA. Results. In a total of 72 FFPE samples of follicular hyperplasias (n = 17), FL (n = 36) and DLBCL (n = 19), SIRT1 staining is localized in the germinal center of the majority of follicular hyperplasias and FL samples. SIRT1 localizes preferentially in the centroblasts of the GC of the follicles where it correlates with Ki67. BCL6 is uniformly positive in follicular hyperplasias and FL, but heterogeneously distributed in DLBCL. Interestingly, SIRT1 and BCL6 expression correlate in FL. By quantitative pyrosequencing we analyzed 3 CpG sites for the SIRT1 promoter (corresponding to the binding sites for CREB, ARID and PPARG transcription factors). Follicular hyperplasias display higher methylation levels (52.88%) than FL (38.36%) and DLBCL (32.65%) on SIRT1 promoter suggesting a possible inverse correlation between tumor aggressiveness and SIRT1 methylation. Next, we selected a panel of genes whose expression is linked to lymphoma pathogenesis. By Methyl II array qPCR, we show that BCL6 methylation does not vary among samples. KLF4, DAPK1 and SPG20, show statistically significant methylation increases in FL and DLBCL compared to follicular hyperplasias, indicating a possible role of these proteins in lymphoma pathogenesis. On the contrary, no significant differences are observed for the other markers MZB1, MGMT, LMO2 and ASXL1. Notably, KLF4, DAPK1 and SPG20 mRNA expression levels anti-correlate with their promoter methylation in FL. Conclusions. Epigenetic changes in SIRT1 methylation inversely correlate with NHL aggressiveness (decreasing in the order: follicular hyperplasias - FL - DLBCL), while KLF4, DAPK1 and SPG20 show a methylation increase that correlates with tumor aggressiveness. These data suggest that different patterns of methylation correlate with the clinical and prognostic parameters of these NHL subtypes. Citation Format: Raffaele Frazzi, Eleonora Zanetti, Mariaelena Pistoni, Ione Tamagnini, Riccardo Valli, Francesco Merli. Different patterns of SIRT1, KLF4, DAPK1 and SPG20 methylation in B lymphocytes correlate with the clinical parameters of non-Hodgkin lymphoma subtypes. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4451.

Unique presentation of a plasmablastic lymphoma superficially involving the entire large bowel.

Plasmablastic lymphoma (PBL) is an uncommon, aggressive B-cell lymphoma mostly occurring in the oral cavity of human immunodeficiency virus (HIV) positive patients, but also described in extraoral sites and in HIV negative patients. One of the relatively common extraoral sites of PBL is the gastrointestinal (GI) tract. Few cases of PBL have been reported in association with inflammatory bowel disease (IBD). Here, we describe the unique presentation of a PBL involving the large bowel superficially along its entire length and without forming a tumor mass in an HIV negative patient with a recent diagnosis of ulcerative colitis.

Epithelioid hemangioma of brachial artery: report of a case and review of the literature

Abstract Epithelioid hemangioma (EH) is an uncommon benign vascular lesion, also known as angioblastic lymphoid (or angiolymphoid) hyperplasia with eosinophilia, characterized by an unclear etiopathogenesis. It usually affects young to middle-aged adults and develops in the head and neck region, as painless cutaneous or subcutaneous reddish papules or nodules. Large vessels involvement is extremely rare, and to date only two cases affecting the brachial artery have been cited in literature. In this report we present a further case of EH of the brachial artery and review the pertinent literature.

Enteropathy-associated T cell lymphoma of the jejunum associated with colonic lymphocytosis with aberrant phenotype

Enteropathy-associated T cell lymphoma is an uncommon variant of T cell lymphoma and it is often associated with coeliac disease (CD) (80–90% of cases).1 A multifocal increase of intraepithelial lymphocytes (IELs) can occur either in the stomach or in the large bowel in the setting of CD, but, to our knowledge, the co-existence of an enteropathy-associated T cell lymphoma of the small bowel and intraepithelial colonic neoplastic low-grade lesions in a patient without CD has not been described.nnA 48-year-old woman came to our attention because of a vague abdominal discomfort. An oesophagogastroduodenoscopy performed to exclude CD turned out to be negative, as did serological testing and HLA DQ2 and DQ8 genotyping. Ten days later, the patient underwent a surgical resection of the jejunum because of intestinal perforation. After diagnosis, staging procedures (bone marrow biopsy, and positron emission tomography and CT scans) were negative for further sites of involvement, and the patient was treated with intensive chemotherapy and autologous stem cell transplantation. Ten months after the diagnosis, the patient was alive, with a poor performance status, but without active disease.nnMacroscopically, the surgical specimen consisted of a tract of jejunum with …