Next-generation sequencing with a myeloid gene panel in core-binding factor AML showed KIT activation loop and TET2 mutations predictive of outcome

Clinical outcome and mutations of 96 core-binding factor acute myeloid leukemia (AML) patients 18-60 years old were examined. Complete remission (CR) after induction was 94.6%. There was no significant difference in CR, leukemia-free-survival (LFS) and overall survival (OS) between t(8;21) (N=67) and inv(16) patients (N=29). Univariate analysis showed hematopoietic stem cell transplantation at CR1 as the only clinical parameter associated with superior LFS. Next-generation sequencing based on a myeloid gene panel was performed in 72 patients. Mutations in genes involved in cell signaling were associated with inferior LFS and OS, whereas those in genes involved in DNA methylation were associated with inferior LFS. KIT activation loop (AL) mutations occurred in 25 patients, and were associated with inferior LFS (P=0.003) and OS (P=0.001). TET2 mutations occurred in 8 patients, and were associated with significantly shorter LFS (P=0.015) but not OS. Patients negative for KIT-AL and TET2 mutations (N=41) had significantly better LFS (P<0.001) and OS (P=0.012) than those positive for both or either mutation. Multivariate analysis showed that KIT-AL and TET2 mutations were associated with inferior LFS, whereas age ⩾40 years and marrow blast ⩾70% were associated with inferior OS. These observations provide new insights that may guide better treatment for this AML subtype.published_or_final_versio

Expression and function of G-protein-coupled receptorsin the male reproductive tract

This review focuses on the expression and function of muscarinic acetylcholine receptors (mAChRs), α1-adrenoceptors and relaxin receptors in the male reproductive tract. The localization and differential expression of mAChR and α1-adrenoceptor subtypes in specific compartments of the efferent ductules, epididymis, vas deferens, seminal vesicle and prostate of various species indicate a role for these receptors in the modulation of luminal fluid composition and smooth muscle contraction, including effects on male fertility. Furthermore, the activation of mAChRs induces transactivation of the epidermal growth factor receptor (EGFR) and the Sertoli cell proliferation. The relaxin receptors are present in the testis, RXFP1 in elongated spermatids and Sertoli cells from rat, and RXFP2 in Leydig and germ cells from rat and human, suggesting a role for these receptors in the spermatogenic process. The localization of both receptors in the apical portion of epithelial cells and smooth muscle layers of the vas deferens suggests an involvement of these receptors in the contraction and regulation of secretion.Esta revisão enfatiza a expressão e a função dos receptores muscarínicos, adrenoceptores α1 e receptores para relaxina no sistema reprodutor masculino. A expressão dos receptores muscarínicos e adrenoceptores α1 em compartimentos específicos de dúctulos eferentes, epidídimo, ductos deferentes, vesícula seminal e próstata de várias espécies indica o envolvimento destes receptores na modulação da composição do fluido luminal e na contração do músculo liso, incluindo efeitos na fertilidade masculina. Além disso, a ativação dos receptores muscarínicos leva à transativação do receptor para o fator crescimento epidermal e proliferação das células de Sertoli. Os receptores para relaxina estão presentes no testículo, RXFP1 nas espermátides alongadas e células de Sertoli de rato e RXFP2 nas células de Leydig e germinativas de ratos e humano, sugerindo o envolvimento destes receptores no processo espermatogênico. A localização de ambos os receptores na porção apical das células epiteliais e no músculo liso dos ductos deferentes de rato sugere um papel na contração e na regulação da secreção.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Universidade Federal de São Paulo (UNIFESP) Escola Paulista de Medicina Departamento de FarmacologiaUNIFESP, EPM, Depto. de FarmacologiaSciEL

Differential gene expression of primitive hematopoietic stem/progenitor cells derived from human umbilical cord blood

Poster no. 284

Clinicopathologic features and outcome of Chinese patients with myelofibrosis

This journal issue contain 2014 ASH Annual Meeting AbstractsINTRODUCTION: Myelofibrosis (MF) is a myeloproliferative neoplasm (MPN) comprising primary myelofibrosis (PMF) and myelofibrosis developing from either polycythemia vera (post-PV MF) or essential thrombocythemia (post-ET MF). METHODS: Two hundred and sixty-seven consecutive patients with MF (PMF, N=206); post-PV MF, N=26; post-ET MF, N=35) diagnosed between January 1988 and December 2013 at seven regional hospitals in Hong Kong were prospectively followed up. Data on the clinicopathologic features and treatment outcome were collected. RESULTS: The median duration of follow-up was 45 (1 – 247) months. The median overall survival (OS) was 65 months (95% confidence interval, CI: 56.3–73.7). The 5-year and 10-year OS were 52.2% and 26.7% respectively. On univariate analysis, factors associated with an inferior OS included age > 65 years (hazard ratio ,HR, =1.81; 95% CI:1.33–2.48; P<0.001), the presence of constitutional symptoms (HR=1.52; 95% CI:1.11–2.08; P=0.009), hemoglobin < 10 g/dL (HR=1.71; 95% CI:1.25–2.35; P=0.001), circulating blasts ≥ 1% (HR=1.55; 95% CI:1.13–2.16; P=0.007), platelet < 100 x 109/L (HR=2.51; 95% CI:1.79–3.53; P<0.001), high risk IPSS (HR=3.09; 95% CI: 1.78–5.37; P<0.001), intermediate-2 risk DIPSS (HR=2.37; 95% CI: 1.38–4.06; P=0.002), high risk DIPSS (HR=2.92; 95% CI:1.51–5.64; P=0.001), transfusion dependence within the first year of diagnosis (HR=2.61; 95% CI:1.92–3.55; P<0.001), transfusion dependence after the first year of diagnosis (HR=2.42; 95% CI:1.61–3.54; P<0.001), palpable hepatomegaly at diagnosis (HR=1.44; 95% CI:1.06–1.96; P=0.02) and secondary AML (HR=1.75; 95% CI:1.24–2.47; P=0.001). On multivariate analysis, post-PV MF (P=0.03), platelet < 100 × 109/L (P=0.001), high risk IPSS (P=0.009), transfusion dependence within the first year (P=0.001), transfusion dependence after the first year (P=0.02), and transformation to secondary acute myeloid leukemia (AML) (P=0.007) were independent risks associated with inferior OS. On univariate analysis, factors associated with increased risk of secondary AML include age ≤ 55 years (odds ratio [OR] = 2.61; 95% CI:1.33–5.12; P=0.005), circulating blasts ≥ 1% (OR=2.24; 95% CI:1.18–4.26; P=0.01), transfusion dependence within the first year (OR=5.57; 95%:2.16–14.88; P<0.001), transfusion dependence after the first year (OR=5.57; 95% CI:2.16–14.88; P<0.001), hepatomegaly (OR=4.05; 95% CI:2.03–8.10), splenomegaly (OR=3.71; 95% CI:1.09–9.26; P=0.04), abnormal karyotypes (OR=3.3; 95% CI:1.14–9.56; P=0.02), and the presence of unfavourable karyotypes (OR=4.9; 95% CI: 1.22–19.96). On multivariate analysis, transfusion dependence within the first year of diagnosis (P=0.04), transfusion dependence after the first year of diagnosis (P=0.003) and hepatomegaly (P=0.006) were independent risks associated with secondary AML. The 5-year and 10-year risks of leukemic transformation were 17.1% and 29.7 respectively. Factors associated with inferior leukemia-free survival (LFS) on univariate analysis included post-ET MF (HR=2.15; 95% CI: 1.03–4.50; P=0.04), presence of constitutional symptoms (HR=1.85; 95% CI:1.04–3.31; P=0.04), circulating blasts ≥ 1% (HR=2.89; 95% CI:1.62–5.16; P<0.001), platelet count < 100 x 109/L (HR=2.56; 95% CI: 1.35–4.84; P=0.004), transfusion dependence within the first year of diagnosis (HR=3.05; 95% CI: 1.70–5.50; P<0.001), transfusion dependence after the first year of diagnosis (HR=6.49; 95% CI: 2.33–18.10; P<0.001), hepatomegaly (HR=3.21; 95% CI:1.69–6.08; P<0.001) and unfavorable karyotype (HR=3.01; 95% CI:1.08–8.35; P=0.03). On multivariate analysis, male gender (P=0.05), presence of constitutional symptoms (P=0.04) and unfavorable karyotypes (P=0.01) were independent risks associated with inferior LFS. Eighteen patients underwent allogeneic haematopoietic stem cell transplantation (HSCT) (matched sibling, N=14; matched-unrelated, N=4), with 15 patients achieving complete remission. Seven patients relapsed with subsequent progression to secondary AML. Acute and chronic graft-versus-host disease occurred in seven (39.9%) and six (33.3%) patients respectively. Transplant-related mortality occurred in three patients. The 5-year and 10-year OS following HSCT was 51.5%. CONCLUSION: Findings of this study complement current prognostic models in guiding treatment decisions at diagnosis and during the course of MF. © 2014 by The American Society of Hematologylink_to_OA_fulltex

Post-genome wide association studies and functional analyses identify association of MPP7 gene variants with site-specific bone mineral density

Our previous genome-wide association study (GWAS) in a Hong Kong Southern Chinese population with extreme bone mineral density (BMD) scores revealed suggestive association with MPP7, which ranked second after JAG1 as a candidate gene for BMD. To follow-up this suggestive signal, we replicated the top single-nucleotide polymorphism rs4317882 of MPP7 in three additional independent Asian-descent samples (n = 2684). The association of rs4317882 reached the genome-wide significance in the meta-analysis of all available subjects (Pmeta = 4.58 × 10-8, n = 4204). Site heterogeneity was observed, with a larger effect on spine than hip BMD. Further functional studies in a zebrafish model revealed that vertebral bone mass was lower in an mpp7 knock-down model compared with the wide-type (P = 9.64 × 10-4, n = 21). In addition, MPP7 was found to have constitutive expression in human bone-derived cells during osteogenesis. Immunostaining of murine MC3T3-E1 cells revealed that the Mpp7 protein is localized in the plasma membrane and intracytoplasmic compartment of osteoblasts. In an assessment of the function of identified variants, an electrophoretic mobility shift assay demonstrated the binding of transcriptional factor GATA2 to the risk allele 'A' but not the 'G' allele of rs4317882. An mRNA expression study in human peripheral blood mononuclear cells confirmed that the low BMD-related allele 'A' of rs4317882 was associated with lower MPP7 expression (P = 9.07 × 10-3, n = 135). Our data suggest a genetic and functional association of MPP7 with BMD variation. © The Author 2011. Published by Oxford University Press. All rights reserved.link_to_subscribed_fulltex