María Teresa Vargas

Tumor microRNA expression profiling identifies circulating microRNAs for early breast cancer detection.

BACKGROUND The identification of novel biomarkers for early breast cancer detection would be a great advance. Because of their role in tumorigenesis and stability in body fluids, microRNAs (miRNAs) are emerging as a promising diagnostic tool. Our aim was to identify miRNAs deregulated in breast tumors and evaluate the potential of circulating miRNAs in breast cancer detection. METHODS We conducted miRNA expression profiling of 1919 human miRNAs in paraffin-embedded tissue from 122 breast tumors and 11 healthy breast tissue samples. Differential expression analysis was performed, and a microarray classifier was generated. The most relevant miRNAs were analyzed in plasma from 26 healthy individuals and 83 patients with breast cancer (36 before and 47 after treatment) and validated in 116 healthy individuals and 114 patients before treatment. RESULTS We identified a large number of miRNAs deregulated in breast cancer and generated a 25-miRNA microarray classifier that discriminated breast tumors with high diagnostic sensitivity and specificity. Ten miRNAs were selected for further investigation, of which 4 (miR-505-5p, miR-125b-5p, miR-21-5p, and miR-96-5p) were significantly overexpressed in pretreated patients with breast cancer compared with healthy individuals in 2 different series of plasma. MiR-505-5p and miR-96-5p were the most valuable biomarkers (area under the curve 0.72). Moreover, the expression levels of miR-3656, miR-505-5p, and miR-21-5p were decreased in a group of treated patients. CONCLUSIONS Circulating miRNAs reflect the presence of breast tumors. The identification of deregulated miRNAs in plasma of patients with breast cancer supports the use of circulating miRNAs as a method for early breast cancer detection.

Application of FISH 7q in MDS patients without monosomy 7 or 7q deletion by conventional G-banding cytogenetics: Does −7/7q− detection by FISH have prognostic value?

Chromosomal abnormalities are detected in 40-60% of patients with de novo myelodysplastic syndromes (MDS). This study used the FISH technique in 773 patients with de novo MDS without evidence of monosomy 7 (-7) or 7q deletion (7q-) by conventional G-banding cytogenetics (CC) to analyze their prognostic impact by FISH alone. FISH detected -7/7q- in 5.2% of patients. Presence of -7/7q- was associated with shorter overall survival than absence of such aberrations. Our results suggest that FISH 7q could be beneficial in patients with intermediate WHO morphologic risk stratification and no evidence of -7/7q- by CC.

Application of fluorescence in situ hybridization as a diagnostic tool in melanocytic lesions, using paraffin wax‐embedded tissues and imprint‐cytology specimens

Background.  Accurate histopathological diagnosis of certain melanocytic skin lesions as benign or malignant can be notoriously difficult. Recently, four‐colour fluorescence in situ hybridization (FISH) has emerged as an important tool for classifying these lesions.

Cytogenetic, fluorescence in situ hybridization, and immunohistochemistry studies in a malignant pleural solitary fibrous tumor.

Pleural solitary fibrous tumor is a normally benign fibroblastic neoplasm; its recurrences and metastasis are associated with clinical and morphological characteristics of variable interpretation and efficacy of surgical treatment. Immunohistochemistry techniques have contributed decisively to the correct diagnosis of the lesion and define its prognosis. In the present case, cytogenetic and fluorescence in situ hybridization analyses revealed multiple chromosomal aberrations, including a del(9)(q21qter) and a marker chromosome ish der(9)(ABL+). The present data support but do not resolve the possible involvement of a gene on 9q22 in the biological behavior of these tumors, and the ABL rearrangements and deletions of 1p and 10p suggest another possible malignant component.

MicroRNA deregulation in triple negative breast cancer reveals a role of miR-498 in regulating BRCA1 expression

Emerging evidence suggests that BRCA1 pathway contributes to the behavior of sporadic triple negative breast cancer (TNBC), but little is known about the mechanisms underlying this association. Considering the central role that microRNAs (miRNAs) play in gene expression regulation, the aim of this study was to identify miRNAs specifically deregulated in TNBC and investigate their involvement in BRCA1 regulation. Using locked nucleic acid (LNA)-based microarrays, expression levels of 1919 miRNAs were measured in paraffin-embedded tissues from 122 breast tumors and 11 healthy breast tissue samples. Differential miRNA expression was explored among the main subtypes of breast cancer, and 105 miRNAs were identified as specific for triple negative tumors. In silico prediction revealed that miR-498 and miR-187-5p target BRCA1, and these results were confirmed by luciferase reporter assay. While miR-187-5p was found overexpressed in a luminal B cell line, miR-498 was highly expressed in a triple negative cell line, Hs578T, and its expression was negatively correlated with the levels of BRCA1. We functionally demonstrated that miR-498 inhibits BRCA1 in breast cancer cell lines, and showed that inhibition of miR-498 led to reduced proliferation in the triple negative cell line Hs578T. Our results indicate that miR-498 regulates BRCA1 expression in breast cancer and its overexpression could contribute to the pathogenesis of sporadic TNBC via BRCA1 downregulation.

Cytogenetic study of a pulmonary sclerosing hemangioma

Pulmonary sclerosing hemangioma (PSH) is an uncommon benign tumor that presents as a solitary asymptomatic and slow-growing nodule. It occurs in both young and old persons; peak incidence is in the fifth decade. Both sexes are affected by this tumor, but women more frequently than men. On histological examination, PSH shows prominent sclerotization and vascularization of the tissue. Recent studies conclude that PSH derives from type II pneumocytes, but the potential for progression and histogenesis remains controversial. We report a case of pulmonary sclerosing hemangioma in a 61-year-old woman with a neoplastic node 1 cm in diameter. The karyotype was 46,XX,t(8;18),der(14;15),+14 in all the cells analyzed. PTEN (10q23) and IgH (14q32) probes were analyzed in interphase nuclei and paraffin-embedded tissues of tumor cells. These chromosome abnormalities could provide information about the relationship of genetic changes to the biological properties of sclerosing hemangioma tumors.

Characterizing patients with multiple chromosomal aberrations detected by FISH in chronic lymphocytic leukemia

Abstract We analyzed the features of chronic lymphocytic leukemia (CLL) with multiple abnormalities (MA) detected by FISH. A local database including 2095 CLL cases was used and 323 with MA (15.4%) were selected. MA was defined by the presence of two or more alterations (deletions of 13q14 (13q-), 11q22 (11q-), 17p13 (17p-) or trisomy 12 (+12)). The combination of 13q- with 11q- and 13q- with 17p-, accounted for 58.2% of the series, in contrast to 11q- with 17p- (3.7%). Patients carrying MA since diagnosis presented a short time to first therapy(TTFT) (27 months) and overall survival (OS) (76 months). The combinations including 17p- had a shorter OS (58 months) than the ones without 17p- (not reached, p = .002). Patients with a complex-FISH were the ones with worse OS (34 months). MA imply poor prognosis when they emerge at diagnosis, probably due to the high incidence of bad prognosis markers, which may be a reflection of a more complex karyotype.

NUP98-HOXA9 bearing therapy-related myeloid neoplasm involves myeloid-committed cell and induces HOXA5, EVI1, FLT3, and MEIS1 expression.

Chromosomal rearrangements involving NUP98 gene have been associated with human leukemias such as de novo AML, therapy‐related AML (t‐AML), myelodysplastic syndrome (MDS), and chronic myeloid leukemia (CML). Genetic fusion NUP98–HOXA9, caused by t(7;11)(p15;p15), is a recurrent cytogenetic alteration in de novo acute myeloid leukemia (AML) usually found in young Asian patients and its description in therapy‐related myeloid neoplasms (t‐MN) is rare. Only one Asian case with molecular demonstration of the NUP98–HOXA9 fusion has been reported in therapy‐related leukemia. NUP98–HOXA9 leukemogenic mechanism is derived from the transcription factor activity of the chimeric protein, which enhances the expression of genes related to cellular differentiation arrest and proliferation.

Methionine synthase reductase deficiency (CblE): A report of two patients and a novel mutation

Importance: Functional methionine synthase reductase deficiency, also known as cobalamin E disorder, is a rare autosomal recessive inherited disease that results in an impaired remethylation of homocysteine to methionine. It presents with macrocytic anemia, hyperhomocysteinemia, and hypomethioninemia, and may also be accompanied with neurological impairment. Clinical presentation: We describe two new cases of unrelated girls with megaloblastic anemia misclassified at first as congenital dyserythropoietic anemia with development of neurologic dysfunction in one of them. Intervention: The posterior finding of biochemical features (hyperhomocysteinemia and hypomethioninemia) focused the diagnosis on the inborn errors of intracellular vitamin B12. Subsequent molecular analysis of the methionine synthase reductase (MTRR) gene revealed compound heterozygosity for a transition c.1361C > T (p.Ser454Leu) and another, not yet described in literature, c.1677–1G > A (p.Glu560fs) in one patient, and a single homozygosis mutation, c.1361C > T (p.Ser545Leu) in the other one. These mutations confirmed the diagnosis of cobalamin E deficiency. Conclusion: Treatment with hydroxocobalamin in combination with betaine appears to be useful for hematological improvement and prevention of brain disabilities in CblE-affected patients. Our study widens the clinical, molecular, metabolic, and cytological knowledge of deficiency MTRR enzyme.

Erythrophagocytosis by peripheral monocytes and histiocytes in diffuse large B cell lymphoma

Dear Editor, An 80-year-old male presented with a 2-month history of weakness, bradypsychia, and fever (38 °C). On physical examination, he was found to be febrile, hypotensive, and tachycardic. No lymphadenopathy was palpable. His full blood count showed hemoglobin concentration 89 g/l, leucocytes 4.63 × 10/l, neutrophils 3.1 × 10/l, monocytes 1.0×10/l, and platelets 65×10/l. Further workup revealed hyperferritinemia (1928 μg/dl), hypertriglyceridemia (352 mg/dl), and elevated soluble CD25 (31.54 ng/ml, normal <7.5). A comprehensive autoimmune and infectious evaluation was negative, including Epstein-Barr virus viral load, serology for hepatitis B, hepatitis C and human immunodeficiency virus. An abdominal and thorax CT scan demonstrated hepatosplenomegaly, with numerous mediastinal and abdominal lymph nodes and pulmonary involvement. Microscopic examination of a blood film revealed monocytes with activation signs such as erythrophagocytosis (Fig. 1a, b, c), pseudopod formation (Fig. 1e, f) and cytoplasm containing a large phagosomal vacuole (Fig. 1g, h, i); even circulating histiocytes showing erythrophagocytosis (Fig. 1d) and prominent vacuolation were identified (Fig. 1j, k). The bonemarrow aspirate was normocellular with adequate global myelopoiesis showing 1 % histiocytes, of which 30 % engulfed erythrocytes, erythroblasts and platelets. The diagnostic criteria for hemophagocytic lymphohistiocytosis (HLH) were met. In addition, a diagnosis for diffuse large B cell lymphoma (DLBCL) was established by fine needle aspiration cytology of a duodenal lymph node (Fig. 2a). Immunohistochemistry revealed a non-germinal centre B cell-like subtype, expressing CD45, CD20, Bcl-6, and MUM-1 with no expression of CD10, or CD3 (Fig. 2b, c, d). The patient died due to respiratory infection 2 days after the bone marrow aspirate. Erythrophagocytosis by monocytes and histiocytes is unusually revealed in a peripheral blood film. This finding has infrequently been described in HLH [1]. It is even rarer if HLH is associated with DLBCL [2]. Hemophagocytosis in peripheral blood may have a worse outcome as has been suggested by the rather few cases reported. Electronic supplementary material The online version of this article (doi:10.1007/s00277-016-2685-z) contains supplementary material, which is available to authorized users.