Comment[ArrayExpressAccession]	E-MTAB-1169
Investigation Title	Autotaxin is expressed in FLT3-ITD positive acute myeloid leukemia and hematopoietic stem cells and promotes cell migration and proliferation
Comment[Submitted Name]	Autotaxin is expressed in FLT3-ITD positive acute myeloid leukemia and hematopoietic stem cells and promotes cell migration and proliferation
Experimental Design	disease_state_design
Experimental Factor Name	AML class	FAB grading
Experimental Factor Type	chromosomal_aberration	tumor_grading
Comment[AEExperimentType]	transcription profiling by array
Comment[AEExperimentDisplayName]	Transcription profiling by array of human acute myeloid leukemia to study ATX expression
Person Last Name	Brors
Person First Name	Benedikt
Person Email	b.brors@dkfz.de
Person Affiliation	German Cancer Research Center Im Neuenheimer Feld 280 69120 Heidelberg
Person Roles	submitter
Quality Control Type	peer_review_quality_control
Public Release Date	2013-05-10
PubMed ID	23377000
Publication Status	submitted
Publication Author List	Claudia Ortlepp, Christine Steudel, Caroline Heiderich, Sina Koch, Angela Jacobi, Martin Ryser, Sebastian Brenner, Martin Bornhäuser, Benedikt Brors, Wolf-Karsten Hofmann, Gerhard Ehninger, Christian Thiede
Publication Title	Autotaxin is expressed in FLT3-ITD positive acute myeloid leukemia and hematopoietic stem cells and promotes cell migration and proliferation
Experiment Description	Autotaxin (ATX) has been reported to act as a motility and growth factor in a variety of cancer cells. The ATX protein acts as a secreted lysophospholipase D (lysoPLD) by converting lysophosphatidylcholine (LPC) to lysophosphatidic acid (LPA), which signals via G-protein coupled receptors and has important functions in cell migration and proliferation. The current study demonstrates that ATX expression is upregulated and functionally active in FLT3-ITD+ human blasts. ATX expression was also found in normal human CD34+ progenitor cells and selected myeloid and lymphoid subpopulations. Stable transduction of mutant FLT3-ITD increased ATX mRNA in selected cell lines, whereas inhibition of FLT3-ITD signaling by sublethal doses of PKC412 led to a significant down-regulation of ATX. Moreover, results indicate that the Jun N-terminal kinase (JNK) is an important mediator between FLT3 signaling and ATX. In the presence of LPC, ATX expression led to a significant increase of proliferation. LPA caused proliferation of all tested cell lines, regardless of ATX expression and induced chemotaxis in human leukemic cell lines and human CD34+ progenitors. LPC increased chemotaxis, in cells with high expression of endogenous and exogenous ATX, by at least 80% demonstrating the autocrine effect of ATX expression. Inhibition of ATX using a small molecule inhibitor induced selective killing of ATX-expressing cell lines and reduced the motile phenotype observed in this cells. Our data suggest that the production of bioactive LPA through ATX is involved in controlling proliferation and migration during hematopoiesis and that deregulation contributes to the pathogenesis of AML. AML Classes: Molecular/cytogenetic group of acute myeloid leukemia, either internal tandem duplication (FLT3) or FLT3 point mutation (D835) or normal karyotype (NK) or t(8;21) transclocation (t821) or monosomy 7 (mono7) or inversion on chromosome 16 (inv16) or high leukocyte count normal karyotype (HL). FAB (French-American-British) Classification system for acute myeloid leukemia is provided for each sample.
Protocol Name	P-MTAB-27282	P-MTAB-27283	P-MTAB-27284	P-MTAB-27285	P-MTAB-27286
Protocol Type	hybridization	nucleic_acid_extraction	feature_extraction	bioassay_data_transformation	labeling
Protocol Description	The generated biotin-labeled cRNA was fragmented by metal-induced hydrolysis, and was hybridized to the GeneChip array (HG-U133A, Affymetrix). The detailed protocol for hybridization, washing and staining of the arrays is available from Affymetrix. Microarray gene expression analysis was performed in triplicates as previously described (Oswald et al., 2006).	High-quality RNA was isolated with the RNeasy Mini Kit (Qiagen, Hilden, Germany) according to the manufacturers instructions. 	After washing and staining, the fluorescence intensity was measured twice for each array using the GeneArray scanner (Affymetrix). 	Fluorescence intensities were normalized to the average fluorescence of the entire microarray. The detailed protocol for further processing of the arrays is available from Affymetrix. 	5 ?g of total RNA were used per patient sample for first strand cDNA synthesis, according to the Eukaryotic Target Labeling protocol as recommended by the manufacturer. Synthesis of biotin-marked cRNA was performed as described in the in vitro-transcription protocol (Affymetrix, Santa Clara, CA, USA).
SDRF File	E-MTAB-1169.sdrf.txt
Publication DOI	10.1016/j.exphem.2013.01.007