Altogen Labs in the Investigation of miRNA expression of Acute Myeloid Leukemia

Publication [Download PDF]: Dmitriy Ovcharenko, Friedrich Stolzel, David Poitz, Fernando Fierro, Markus Schaich, Andreas Neubauer, Kevin Kelnar, Timothy Davison, Carsten Muller-Tidow, Christian Thiede, Martin Bornhauser, Gerhard Ehninger, David Brown, and Thomas Illmer. Experimental Hematology 2011; 39:1030–1042

Acute myeloid leukemia (AML) is a cancer that affects hematopoietic stem cells and often results in the accumulation of immature or abnormal blood cells that interfere with healthy blood cell function. AML patients are categorized and treated differently based on cytogenetic alterations; AML blast chromosomal aberrations have been linked to relapse or treatment failure. While intermediate risk (IR) AML patience have a normal karyotype, this is often accompanied by additional mutations, for example in FLT3, CEBP/α, or NPM1, which can be used to predict prognosis and treatment response. Methods for discriminating subtypes of AML are critical to successful treatment, and Ovcharenko et al. (Experimental Hematology, 2011) analyzed differential expressions of microRNA (miRNA) in AML/IR patients. Chromosomal abnormalities are known to alter miRNA expression levels, which in turn regulate gene expression levels.  Specifically in hematopoietic cells, miRNAs have been seen to regulate differentiation and proliferation. The sensitive nature of miRNA expression profiles may prove to have a higher sensitivity than karyotyping or complementary DNA microarrays for characterizing AML subtypes.

With services provided by Altogen Labs, the 2011 Experimental Hematology study identified miR-10a, a miRNA that is upregulated in AML nucleophosmin 1 (NPM1) mutant samples, and analyzed in vitro functional consequences of this effect. Microarray analysis and quantitative reverse transcription polymerase chain reaction (qRT-PCR) of miRNAs were performed to compare normal to AML IR patient samples and led to the identification of miR-10a as upregulated in ML samples with mutant NPM1. qRT-PCR, western blotting, proliferation assays, overexpression studies, anti-miR-10a inhibition studies and mutant studies were used to analyze the potential effect of miR-10a cellular gene expression and signaling. Overall, data suggested that the characteristic overexpression of miR-10a in NPM1-mutant AML patients may result in cellular changes mediated via murine double minute 4 (MDM4) and the p53 signaling pathway. Altogen Laboratories is Texan contract research organization (CRO) offering services such as those outlined above and more. Altogen Labs is equipped to provide RNAi services (siRNA encapsulation, stable gene knockdown, transient transfection, shRNA construction, inducible knockdown, etc.), stable protein overexpression cell line, cryopreservation cell banking, IC50 toxicology studies, a wide array of xenograft studies and more. Professional scientists at Altogen Labs are trained to provide top quality research at competitive pricing. At Altogen Labs we understand time is important, which is why we offer instant quotes as well as any desired consulting throughout the research process, from experiment design to analysis, in order to provide clients with a seamless and efficient experience.