Mar García

MicroRNA expression profiling and DNA methylation signature for deregulated microRNA in cutaneous T-cell lymphoma

MicroRNAs usually regulate gene expression negatively, and aberrant expression has been involved in the development of several types of cancers. Microarray profiling of microRNA expression was performed to define a microRNA signature in a series of mycosis fungoides tumor stage (MFt, n=21) and CD30+ primary cutaneous anaplastic large cell lymphoma (CD30+ cALCL, n=11) samples in comparison with inflammatory dermatoses (ID, n=5). Supervised clustering confirmed a distinctive microRNA profile for cutaneous T-cell lymphoma (CTCL) with respect to ID. A 40 microRNA signature was found in MFt including upregulated onco-microRNAs (miR-146a, miR-142-3p/5p, miR-21, miR-181a/b, and miR-155) and downregulated tumor-suppressor microRNAs (miR-200ab/429 cluster, miR-10b, miR-193b, miR-141/200c, and miR-23b/27b). Regarding CD30+ cALCL, 39 differentially expressed microRNAs were identified. Particularly, overexpression of miR-155, miR-21, or miR-142-3p/5p and downregulation of the miR-141/200c clusters were observed. DNA methylation in microRNA gene promoters, as expression regulatory mechanism for deregulated microRNAs, was analyzed using Infinium 450K array and approximately one-third of the differentially expressed microRNAs showed significant DNA methylation differences. Two different microRNA methylation signatures for MFt and CD30+ cALCL were found. Correlation analysis showed an inverse relationship for microRNA promoter methylation and microRNA expression. These results reveal a subgroup-specific epigenetically regulated microRNA signatures for MFt and CD30+ cALCL patients.

First-line response-adapted treatment with the combination of bendamustine and rituximab in patients with mucosa-associated lymphoid tissue lymphoma (MALT2008-01): a multicentre, single-arm, phase 2 trial

BACKGROUND No standard first-line systemic treatment for mucosa-associated lymphoid tissue (MALT) lymphoma is available. In a phase 2 study we aimed to assess the safety and activity of a response-adapted combination of bendamustine plus rituximab as upfront treatment for this type of lymphoma. METHODS In a multicentre, single-arm, non-randomised, phase 2 trial, we enrolled patients with MALT lymphoma at any site and stage and treated them with bendamustine (90 mg/m(2) on days 1 and 2) plus rituximab (375 mg/m(2) on day 1), every 4 weeks. Inclusion criteria were measurable or evaluable disease, age 18-85 years, and unequivocal active lymphoma; we also enrolled patients with MALT lymphoma arising in the stomach after failure of Helicobacter pylori eradication and primary cutaneous cases after failure of local therapies. Exclusion criteria included evidence of histological transformation, CNS involvement, and active hepatitis B or C virus or HIV infection. After three cycles, patients achieving complete response received one additional cycle (total four cycles) and those achieving partial response received three additional cycles (total six cycles). The primary endpoint was 2-year event-free survival. Analysis was by modified intention to treat. This trial is registered with ClinicalTrials.gov, number NCT01015248. FINDINGS 60 patients from 19 centres in Spain were enrolled between May 27, 2009, and May 23, 2011, and received treatment; 57 patients were evaluable for the primary endpoint. Only 14 (25%) patients needed more than four cycles of treatment. After a median follow-up of 43 months (IQR 37-51), median event-free survival was not reached. Event-free survival at 2 years was 93% (95% CI 84-97) and at 4 years was 88% (95% CI 74-95). The most frequently observed grade 3-4 adverse events were haematological: lymphopenia in 20 (33%) patients, neutropenia in 12 (20%) patients, and leucopenia in three (5%) patients. Grade 3-4 febrile neutropenia or infections were reported in three (5%) and four (7%) patients, respectively. INTERPRETATION This response-adapted schedule of bendamustine plus rituximab appears to be an active and well tolerated first-line treatment for patients with MALT lymphoma. FUNDING Grupo Español de Linfomas/Trasplante de Médula Ósea (GELTAMO), Mundipharma Spain, and Roche Pharma Spain.

Study of Regulatory T-Cells in Patients with Gastric Malt Lymphoma: Influence on Treatment Response and Outcome

Purpose FOXP3+ regulatory T cells (Treg) play an essential role in modulating host responses to tumors and infections. The role of these cells in the pathogenesis of MALT lymphomas remains unknown. The aims of the study were to quantify the number of infiltrating FOXP3+ and CD3+ cells in patients with gastric MALT lymphoma at diagnosis and to study kinetics of these cells and CD20+ tumor cells after treatment and during long-term follow-up. Methods FOXP3+, CD3+ and CD20+ cells were analyzed by immunohistochemistry and the number of cells was quantified using a micrometric ocular. Samples of 35 patients with gastric MALT lymphoma at diagnosis and after treatment were included. Diagnostic samples were compared to 19 cases of chronic gastritis and diffuse large B-cell lymphoma (DLBCL) of the stomach. Results The median number of FOXP3+ infiltrating cells was higher (27 cells/cm2) in gastric MALT patients than in DLBCL (10 cells; p = 0.162) but similar to chronic gastritis (20 cells; p = 0.605). No characteristic or specific distribution pattern of infiltrating FOXP3+ cells was found. Gastric MALT lymphoma patients responding to bacterial eradication therapy had higher number of FOXP3+ cells at study entry. Kinetics of both infiltrating FOXP3+ cells and tumor CD20+ cells were strongly dependent on the treatment administered. Discussion Gastric MALT lymphomas have a number of Treg cells more similar to chronic gastritis than to DLBCL. Patients with higher number of tumor infiltrating FOXP3+ cells at study entry seem to have better response to antibiotics. Kinetics of Treg and tumor cells are influenced by type of treatment.

Linfoma primario de colon

BACKGROUND primary colorectal lymphoma is a very rare disease, representing less than 0.5 % of all primary colorectal neoplasms. The gastrointestinal tract is the most frequently involved site of all extranodal lymphomas, the most common type of that is non-Hodgkin s lymphoma. Early diagnosis is often difficult because of unspecific symptoms. Therapeutic approaches have classically included radical resection, chemotherapy and radiotherapy. MATERIALS AND METHODS we present our experience in the management of primary colorectal lymphomas over a 17-year period (1994-20011). RESULTS in this period 7 cases of primary colorectal lymphoma were diagnosed in our institution. Abdominal pain and change in bowel habit were the most frequent symptoms. Five patients underwent emergency surgery because of bleeding or bowel obstruction. All primary intestinal lymphomas studied were of the Bcell phenotype. Patients were followed up for a median of 59 months (range 1-180). Three of them are alive with no evidence of recurrence. CONCLUSION combination treatment with chemotherapy and surgery can obtain good remission rate. Surgery can resolve complications such bleeding or intestinal perforation that are implicated in lymphoma mortality.

Cryptic IGH/BCL2 rearrangements with variant FISH patterns in follicular lymphoma

Follicular lymphoma (FL) is one of the most common non-Hodgkin lymphomas (NHL). Translocation t(14;18)(q32;q21) involving IGH and BCL2 genes represents its genetic hallmark. We present six cases of a series of 75 well diagnosed FL patients in which variant fluorescence in situ hybridization (FISH) patterns for this rearrangement were found. Moreover, G-banding cytogenetics and polymerase chain reaction (PCR) methods were unable to detect t(14;18)(q32;q21). According to our results, FISH is the best technique to define variant rearrangements of IGH/BCL2 genes and is important to detect it in cases with non-conclusive FL characteristics to avoid misdiagnosis with other NHL.

Pathology reporting of bone marrow biopsy in myelofibrosis; application of the Delphi consensus process to the development of a standardised diagnostic report

Aims The diagnosis of primary myelofibrosis (PMF) strongly relies on the bone marrow biopsy findings, but a report model has not been standardised. Our aim was to establish general recommendations for bone marrow evaluation and standardised reporting in a case suspicious of PMF. Methods The Delphi method was employed to obtain expert consensus. An advisory panel of 10 leading members identifies a total of 37 haematopathology experts to participate. The first Delphi round included a questionnaire with three main groups of items: minimal clinical and laboratory data considered necessary before reporting, minimal descriptive aspects to record and main histological differential diagnosis. The final report content was based on consensus obtained after the second Delphi round. Results The minimal data considered necessary were age, splenomegaly, haemoglobin, leucocyte and platelet counts, differential blood cell count, leucoerythroblastic blood picture, lactate dehydrogenase (LDH) level, BCR-ABL and JAK2 mutational status, reticulin stain and the internal control for the reticulin staining. The minimal descriptive aspects to report were cellularity, osteosclerosis, megakaryocytic morphology and localisation, dense megakaryocytic clusters, quantity of granulocytic precursors, grade of myelofibrosis in a scale of 4, and a proposed final diagnostic approach. The entities to be considered for differential diagnosis were mainly the other classical chronic myeloproliferative neoplasms. Conclusions The Delphi method is a robust tool to determine essential information to be included in a pathology report. A standardised good-quality histopathological report form may help to homogenise PMF diagnosis. A close collaboration between the pathologist and the haematologist is desirable according to our survey.

Bendamustine-containing immunochemotherapy is active in transformed follicular lymphoma with overexpression of p53

Bendamustine is a purine analogue/alkylator hybrid [1] that has demonstrated clinical activity in patients with relapsed indolent non-Hodgkin lymphoma (NHL), including those refractory to other alkylating or purine analogue agents [2]. Bendamustine has been investigated both as a single agent and in combination with monoclonal antibodies in patients with NHL [3,4]. Recently, Friedberg et al. [5] reported the clinical activity of bendamustine in patients with rituximab-refractory indolent and transformed non-Hodgkin lymphoma from phase II multicentre study. In this report, bendamustine as single-agent produced a high rate of responses with acceptable toxicity in this heavily pretreated group of patients. Of note, bendamustine achieved responses even in patients with transformed disease, although time to progression was relatively short (p53 mutation data not shown). We report a case of transformed NHL with overexpression of p53 who remained in complete remission with bendamustine-containing chemotherapy after 6 months. A 70-year-old female patient with stage II NHL, follicular small cleaved type, was diagnosed in May 1993 when she presented with inguinal lymph nodes. Computed tomography (CT) scan revealed a bulky retroperitoneal mass and inguinal lymphadenopathy. Bone marrow was not involved. The patient received chemotherapy with six cycles of CHOP and radiotherapy to the bulky mass, achieving complete remission. Six years later, the patient relapsed in a left-side cervical node and, after local radiotherapy, she went into a second complete remission. The patient remained free of disease for 4 years but she had a second relapse with pleural effusion, lymphadenopathy at multiple regions and bone marrow involvement. She was treated with six cycles of FCM and achieved a third complete remission. Peripheral stem cell harvest was tried but stem cells could not be collected and maintenance with anti-CD20 monoclonal antibody (rituximab) was started. She relapsed again one-and-a-half years after starting rituximab maintenance. Four cycles of R-GEMOX were administered and she achieved a partial response, but a further progressive lymphoma was seen 3 months later. A lymph node biopsy showed large cell lymphoma transformation with immunohistochemistry positive for CD20 and overexpression of p53. Then she was treated with four cycles of modified R-CHOP (with non-pegylated liposomal doxorubicin) followed by a consolidation with 0.4 mCi/kg of 90 ibritumomab tiuxetan (Zevalin) but she progressed 3 months later. At this time, we considered treatment with bendamustine and she received six cycles of R-BOP [6] (rituximab 375 mg/m day 1, bendamustine 90 mg/m daily on days 1–2, vincristine 2 mg on day 1 and prednisone 100 mg daily on days 1–5). Tolerance was excellent, the main toxicities being vincristine-associated neurotoxicity of grade 1 and mild neutropenia that was treated with pegfilgastrim in the last two cycles in order to maintain the dose intensity. At the last

Long-term results of a phase 2 study of rituximab and bendamustine for mucosa-associated lymphoid tissue lymphoma

To the editor: Extranodal marginal zone B-cell lymphomas of mucosa-associated lymphoid tissue (MALT) present in most cases as localized disease and can be successfully treated with local therapies or antibiotics.[1][1][⇓][2][⇓][3]-[4][4] Moreover, patients with multicentric or extensive disease

Diagnostic Accuracy of the Enhanced Liver Fibrosis (ELF®) Score Using HCV-Infected Serum Samples Cryopreserved for up to 25 Years

Introduction & Aims Cryopreservation of serum samples is a standard procedure for biomedical research in tertiary centers. However, studies evaluating the long-term biological stability of direct liver fibrosis markers using cryopreserved samples are scarce. Methods We compared the stability of hyaluronic acid (HA), tissue inhibitor of metalloproteinases (TIMP-1) and amino-terminal propeptide of type III procollagen (PIIINP) in 225 frozen serum samples of HCV-infected patients with a paired liver biopsy for up to 25 years (1990–2014). Moreover, we assessed the diagnostic accuracy (AUROC) of the Enhanced Liver Fibrosis (ELF®) score to identify significant fibrosis (F2-4) and its predictive capacity to identify clinical events during follow-up. Results Seventy-six patients (39,8%) had mild fibrosis (F0-1) and 115 (60,2%) significant fibrosis (F2-4). HA, PIIINP and TIMP-1 values remained stable during the period from 1995 to 2014 while those of 1990–94 were slightly higher. We did not find significant differences in the median ELF® values during the 20-year period from 1995–2014 in patients with mild (from 8,4 to 8,7) and significant fibrosis (from 9,9 to 10,9) (p = ns between periods and fibrosis stages). The AUROCs of ELF® to identify significant fibrosis were high in all the periods (from 0,85 to 0,91). The ELF® score showed a good predictive capability to identify clinical events during follow-up. Conclusions The biological stability of direct serum markers (HA, PIIINP and TIMP-1) using HCV-infected samples cryopreserved for 20 years is good. Therefore, the diagnostic accuracy of the ELF® score to identify significant fibrosis and clinical events during follow-up is very high.

Cutis laxa-like mycosis fungoides

Mycosis fungoides is the most common form of primary cutaneous T‐cell lymphoma. Several clinical and clinicopathological variants of mycosis fungoides have been reported. A 75‐year‐old woman presenting with multiple ill‐defined areas of marked cutaneous wrinkling on the trunk and extremities is reported. Histopathological examination showed characteristic features of mycosis fungoides along with an interstitial dermal infiltrate composed predominantly of atypical lymphocytes with histiocytes intermingled within the collagen bundles. A focal reduction and fragmentation of elastic fibers was demonstrated. This observation illustrates a peculiar and previously unreported clinicopathological presentation of mycosis fungoides: cutis laxa‐like mycosis fungoides, expanding the spectrum of mycosis fungoides variants associated with abnormalities of the dermal elastic fibers.