Unknown,Transcriptomics,Genomics,Proteomics

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MiRNA expression in AML and healthy controls

ABSTRACT: Acute myeloid leukemia (AML) is a hematopoietic malignancy with a dismal outcome in the majority of cases. A detailed understanding of the genetic alterations and gene expression changes that contribute to its pathogenesis is important to improve prognostication, disease monitoring, and therapy. The expression of 636 human miRNAs was compared between samples from 52 patients with AML and 13 healthy individuals by locked nucleic acid (LNA) based microarray technology. 143 miRNAs were expressed at detectable levels, and 64 of these were significantly differentially expressed between AML and healthy peripheral blood, bone marrow, and/or CD34+ cells. Reference: A Rommer et al, Overexpression of primary microRNA 221/222 in acute myeloid leukemia, BMC Cancer, 2013. 52 AML, 5 peripheral blood (PB), 5 bone marrow (BM), and 3 CD34+ cell samples from healthy donors were subjected to miRNA microarray analysis.

ORGANISM(S): Homo sapiens

SUBMITTER: Rotraud Wieser

PROVIDER: E-GEOD-49665 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Overexpression of primary microRNA 221/222 in acute myeloid leukemia.

Rommer Anna A Steinleitner Katarina K Hackl Hubert H Schneckenleithner Christine C Engelmann Maria M Scheideler Marcel M Vlatkovic Irena I Kralovics Robert R Cerny-Reiterer Sabine S Valent Peter P Sill Heinz H Wieser Rotraud R

BMC cancer 20130729

<h4>Background</h4>Acute myeloid leukemia (AML) is a hematopoietic malignancy with a dismal outcome in the majority of cases. A detailed understanding of the genetic alterations and gene expression changes that contribute to its pathogenesis is important to improve prognostication, disease monitoring, and therapy. In this context, leukemia-associated misexpression of microRNAs (miRNAs) has been studied, but no coherent picture has emerged yet, thus warranting further investigations.<h4>Methods</ ...[more]

PMID: 23895238

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Project description:MicroRNAs (miRNAs) have been shown to play an important role in many different cellular, developmental, and physiological processes. Accordingly, numerous methods have been established to identify and quantify miRNAs. The shortness of miRNA sequence results in a high dynamic range of melting temperatures and, moreover, impedes a proper selection of detection probes or optimized PCR primers. While miRNA microarrays allow for massive parallel and accurate relative measurement of all known miRNAs, they have so far been less useful as an assay for absolute quantification. Here, we present a microarray based approach for global and absolute quantification of miRNAs. The method relies on an equimolar pool of about 1000 synthetic miRNAs of known concentration which is used as an universal reference and labeled and hybridized in a dual colour approach on the same array as the sample of interest. Each single miRNA is quantified with respect to the universal reference outbalancing bias related to sequence, labeling, hybridization or signal detection method. We demonstrate the accuracy of the method by various spike in experiments. Further, we quantified miRNA copy numbers in liver samples and CD34(+)CD133(-) hematopoietic stem cells. The RNA extracted from 7 x 10^ 5 to 1 x 10^ 6 CD34(+)/CD133(-) cells of three different donors was analyzed. 1 µg of respective total RNA was mixed with 2.5 fmol of each of 18 RNA oligonucleotides reverse complement to miRControl 3 probes and subsequently fluorescently labelled by 3’ ligation. Total RNA mix was hybridized in a dual colour approach to microarrays versus a second labelled synthetic miRNA pool. The synthetic miRNA pool consisted of 2.5 fmol of each of 954 non redundant miRNAs sequences and miRControl 3 sequences. The array data was normalized by calculating the median of the miRControl 3 present in the CD34(+)/CD133(-) and UR sample. The miRNA amount was calculated with respect to the corresponding miRNA in the UR.

Dataset Information

MiRNA expression in AML and healthy controls

Publications

Overexpression of primary microRNA 221/222 in acute myeloid leukemia.

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