Comment[ArrayExpressAccession] E-GEOD-25311 MAGE-TAB Version 1.1 Public Release Date 2011-03-24 Investigation Title MicroRNAs are Transported in Plasma and Delivered to Recipient Cells by High-Density Lipoproteins (HG-U133 2.0) Comment[Submitted Name] MicroRNAs are Transported in Plasma and Delivered to Recipient Cells by High-Density Lipoproteins (HG-U133 2.0) Experiment Description Circulating microRNAs (miRNA) are relatively stable in plasma and are a new class of disease biomarkers. Here we present evidence that human high-density lipoprotein (HDL) transports endogenous miRNAs and delivers them to recipient cells with functional targeting capabilities. Highly-purified fractions of human HDL contain small RNAs, and the HDL-miRNA profile from normal subjects is significantly different than familial hypercholesterolemia subjects. miRNAs were demonstrated to associate with both native and reconstituted HDL particles, and reconstituted HDL injected into mice retrieved distinct miRNA profiles from normal and atherogenic models. Cellular export of miRNAs to HDL was demonstrated to be regulated by neutral sphingomyelinase. HDL-mediated delivery of miRNAs to recipient cells was demonstrated to be scavenger receptor BI-dependent. Furthermore, HDL delivery of both exogenous and endogenous miRNAs resulted in the direct targeting of mRNA reporters. Notably, HDL-miRNA from atherosclerotic subjects induced differential gene expression, with significant loss of conserved mRNA targets in cultured hepatocytes. Collectively, these observations suggest that HDL participates in a novel mechanism of intercellular communication involving the transport and delivery of miRNAs. Gene expression changes in human Huh7 cells with familial hypercholesterolemia HDL treatment. Gene expression (mRNA) profiles in human Huh7 cells treated with normal HDL (n=3) or FH HDL (n=3) in lipoprotein-depleted serum (48h). Term Source Name ArrayExpress EFO Term Source File http://www.ebi.ac.uk/arrayexpress/ http://www.ebi.ac.uk/efo/efo.owl Person Last Name Vickers Vickers Palmisano Shoucri Shamburek Remaley Person First Name Kasey Kasey Brian Bassem Robert Alan Person Mid Initials C C T M D T Person Email vickerskc@nhlbi.nih.gov Person Affiliation National Institutes of Health Person Phone 301-496-5114 Person Fax 301-402-0190 Person Address National Heart, Lung and Blood Institute, National Institutes of Health, 10 Center Dr. Bldg. 10 8N222, Bethesda, MD, USA Person Roles submitter Protocol Name P-GSE25311-1 P-GSE25311-6 P-GSE25311-3 P-GSE25311-8 P-GSE25311-7 P-GSE25311-2 P-GSE25311-4 P-GSE25311-5 Protocol Description ID_REF = VALUE = LiWong (MBEI) summarized signal intensity. Following fragmentation, 15ug cRNA were hybridized 16 h @ 42oC to Affymetrix HG-U133 Plus 2.0 human whole genome microarrays. Arrays were washed and stained with Affymetrix Fluidics Station. Huh7 cells were cultured in normal growth media (F12 Dulbecco’s modified Eagle’s medium, supplemented with 100U/mL penicillin, 100ug/mL streptomycin, and 10% fetal bovine serum (FBS). Low-level and high-level post-analyses of mRNA (LiWong method) expression were performed using GeneSpring GX11.0 (Agilent Technologies). LiWong summarization algorithm, quanitle normalization, no baseline transforamtion, filtered on expression (20.0-100.0)th percentile, and filtered on error CV<50.0%. Data presented here: all entities and summarization signal values Arrays were scanned using the Affymetrix GeneChip Scanner Huh7 cells were washed with warm 1X PBS and switched to a lipoprotein-depleted serum (10%) media prior to treatment. Cells were treated with 70ug/mL HDL for 48 h. Cells were washed 2X with 1X PBS and lysed with Qiazol Total RNA was isolated by Qiazol extraction followed by miRNAeasy Column (Qiagen) purification Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 1 ug total RNA Protocol Type bioassay_data_transformation hybridization grow feature_extraction image_aquisition specified_biomaterial_action nucleic_acid_extraction labeling Experimental Factor Name TREATMENT Experimental Factor Type treatment Publication Title MicroRNAs are transported in plasma and delivered to recipient cells by high-density lipoproteins. Publication Author List Vickers KC, Palmisano BT, Shoucri BM, Shamburek RD, Remaley AT PubMed ID 21423178 Publication DOI 10.1038/ncb2210 Comment[SecondaryAccession] GSE25311 Comment[GEOReleaseDate] 2011-03-24 Comment[ArrayExpressSubmissionDate] 2010-11-12 Comment[GEOLastUpdateDate] 2013-01-14 Comment[AEExperimentType] transcription profiling by array SDRF File E-GEOD-25311.sdrf.txt