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Management of systemic mastocytosis is an emerging challenge which requires a multidisciplinary diagnostic approach and personalized treatment strategy. Midostaurin can rapidly reduce the disease burden, also in cladribine refractory cases.

In spite of an increasing array of investigations, the relationships between viral infections and allogeneic hematopoietic stem cell transplantation (HSCT) are still controversial, and almost exclusively regard DNA viruses. Viral infections per se account for a considerable risk of morbidity and mortality among HSCT recipients, and available antiviral agents have proven to be of limited effectiveness. Therefore, an optimal management of viral infection represents a key point in HSCT strategies. On the other hand, viruses bear the potential of shaping immunologic recovery after HSCT, possibly interfering with control of the underlying disease and graft-versus-host disease (GvHD), and eventually with HSCT outcome. Moreover, preliminary data are available about the possible role of some virome components as markers of immunologic recovery after HSCT. Lastly, HSCT may exert an immunotherapeutic effect against some viral infections, notably HIV and HTLV-1, and has been considered as an eradicating approach in these indications.

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7563Background: Metronomic chemotherapy (MC) is an emerging approach in solid tumours. Since MC has been poorly investigated in aggressive lymphomas in 2011, we formulated a new oral MC regimen ter...

Amyloidosis is a potentially fatal condition characterized by deposition of extracellular protein in an abnormal fibrillary form with a β-sheet structure, named amyloid, in several organs, especially in heart, kidney and liver. The tissue biopsy, stained with Congo red and demonstrating amyloid deposits with apple-green birefringence, is required for diagnosis. The biopsy of visceral organs is characterized by higher sensitivity although it requires invasive procedures bearing higher risk of complications, such as bleeding, hematoma and perforation. Therefore, fine-needle abdominal fat aspiration is the most common biopsy site. More recently an additional procedure has been performed and is at this time is under evaluation, being represented by minor salivary labial glands biopsy (MSGB) which is characterized by easy accessibility, low complication rate, and lower costs. Reliability of this procedure is under evaluation. We have analyzed all patients referred at our institution between March 2006 and April 2015, with the clinical or laboratory symptoms suggestive for amyloidosis (proteinuria, renal impairment, neuropathy and restrictive cardiomyopathy). In the first three years we performed biopsy by abdominal fat aspiration as first diagnostic step. Patients failing to obtain diagnostic material underwent a second biopsy, including surgical approaches. In order to minimize the use of invasive procedures, we have introduced MSGB, obtained by a small incision inside the lower lip. The sample was then collected and fixed in formalin, stained with Congo red and analyzed by polarized light microscopy. Immunohistochemical examination was performed whenever indicated to discriminate AA and AL amyloidosis. In our retrospective study we have examined results of MSGB from 44 patients during the time period between January 2008 and April 2015. All patient were affected with AL amyloidosis and 51% of them was associated with multiple myeloma. The deposits were characterized as AL λ in 80% of the patients and AL κ in 20%. The median age was 65 years (range 26-97), 30% were female and 70% male. In 38 patients (86%) the MSGB was positive, while in six (14%) was negative. Of these six patients, two presented localized amyloidosis (diagnosed by tongue and conjunctiva biopsy) and four underwent visceral organ biopsy (3 renal biopsies, 1 myocardial). MSGB is therefore a simple, safe, and reliable tool for the diagnosis of systemic amyloidosis. The advantages of MSGB include avoidance of more invasive methods, need for a small incision, quite a low risk of bleeding and nerve damage, applicability in the outpatient setting and rapid healing. In our experience, it was performed in all cases without adverse effects, demonstrating an overall diagnostic sensitivity of 86%. For this reason we introduce this procedure in our clinical practice. Disclosures No relevant conflicts of interest to declare.

Chronic myelomonocytic leukemia (CMML) represents a diagnostic and therapeutic challenge characterized by highly heterogeneous clinical and laboratory aspects, contrasting from mainly dysplastic (MD) to predominantly proliferative (MP) in different patients. Although no specific cytogenetic or molecular aberration has been associated to CMML, next generation sequencing (NGS) has recently led to the discovery of at least one lesion in up to 90% of patients. Nonetheless, the role of the identified genetic aberrations in CMML onset and progression remains to be clarified. In a series of 40 consecutive patients we previously reported a higher frequency of RAS and JAK2 mutations and a shorter survival in those with MP- than in those with MD-disease. Furthermore, paired samples analysis showed RAS mutations acquisition in concomitance with progression from MD- to MP-CMML, suggesting these lesions as second hits that confer a proliferative advantage to the malignant clone, leading to poor outcome. In addition to these findings, a highly significant shorter life expectation in the MP-variant of CMML was more recently confirmed in an extended population of 74 patients (p=0.0005), further supporting the association of molecular acquisition of gene aberrations with disease progression. By comprehensive next generation sequencing (NGS) of selected genes, here we aimed to further investigate the spectrum of aberrations contributing to CMML development and progression and to examine whether MD- and MP-CMML may be also discriminated at the molecular level. We designed a NGS study (Oxford Gene Technology, Oxford UK) of 44 genes in DNA prepared from MNCs from 12 CMML patients after obtaining informed consent. Of the 21 samples analyzed, 17 were consecutively collected from 9 patients at the time of MD-CMML and later on during the disease course, showing either long lasting stable MD-CMML disease (median follow-up of 102 month), or progression to MP-CMML or AML, and 4 more were obtained from patients with MP-CMML (2 with previous MD-phase). In some patients, DNA prepared from purified CD3+ cells selected by FACS cell sorting was also analyzed. Candidate mutations were validated by Sanger sequencing. Deep sequencing analysis confirmed TET2 mutations as the most frequent (10/12 patients, 83%) and, the earliest known event in CMML, being present since time of referral in 100% of our cases with sequential samples, supporting their possible role of initiating lesions in CMML. Overall, 9 patients harbored frameshift/nonsense mutations and 1 had an essential splice site substitution. Non-synonymous variations of yet unknown origin were detected in 3 cases while in 1 case the substitution found in MNCs DNA was identified by direct sequencing also in DNA from buccal swab and thus annotated as a SNP. Other documented mutations in variable proportions involved ASXL1, SRSF2, SF3B1, EZH2, CBL, DNMT3A, MPL, NOTCH1, NOTCH2, N- and K-RAS. Among patients who were investigated with sequential samples collected at different time points and/or different disease phases, TET2, SRSF2 and ASXL1 mutations were documented from the first presentation in all cases, suggesting their acquisition as early events possibly driving molecular mechanisms of disease onset. In contrast, besides RAS mutations, which were detected at the time of disease progression from the MD- to the MP-variant in 2 patients, other aberrations possibly associated with disease evolution included EZH2 and CBL mutations, both detected in a small fraction of cells at diagnosis but significantly expanding after progression to MP-CMML. Of note, in one case harboring TET2, ASXL1, EZH2 and CBL concomitant mutations the sequencing of DNA from purified CD3+ cells unveiled the presence of TET2, ASLX1 and CBL mutations also in a significant fraction of T-lymphocytes, suggesting the aberration to possibly arise in a multipotent progenitor, whereas the EZH2 mutation appeared restricted to the myeloid lineage. A combined analysis of sequential samples and single-cell-derived colonies is currently ongoing to better elucidate clonal evolution in CMML, which in turn could help the improvement of disease classification as well as the early identification of patients at risk of disease evolution. Disclosures: No relevant conflicts of interest to declare.