Comment[ArrayExpressAccession]	E-GEOD-57718
MAGE-TAB Version	1.1							
Public Release Date	2015-02-20
Investigation Title	Pi3kcb links Hippo-YAP and PI3K-AKT signaling pathways to promote cardiomyocyte proliferation and survival [microarray]							
Comment[Submitted Name]	Pi3kcb links Hippo-YAP and PI3K-AKT signaling pathways to promote cardiomyocyte proliferation and survival [microarray]
Experiment Description	Background—YAP, the nuclear effector of Hippo signaling, regulates cellular growth and survival in multiple organs, including the heart, by interacting with TEAD sequence specific DNA-binding proteins. Recent studies showed that YAP stimulates cardiomyocyte proliferation and survival. However, the direct transcriptional targets through which YAP exerts its effects are poorly defined.   Methods and Results—To identify genes directly regulated by YAP in cardiomyocytes, we combined differential gene expression analysis in YAP gain- and loss-of-function with genome-wide identification of YAP bound loci using chromatin immunoprecipitation and high throughput sequencing. This screen identified Pik3cb, encoding p110β, a catalytic subunit of phosphoinositol-3-kinase (PI3K), as a candidate YAP effector that promotes cardiomyocyte proliferation and survival. We validated YAP and TEAD occupancy of a conserved enhancer within the first intron of Pik3cb, and show that this enhancer drives YAP-dependent reporter gene expression. Yap gain- and loss-of-function studies indicated that YAP is necessary and sufficient to activate the PI3K-Akt pathway. Like Yap, Pik3cb gain-of-function stimulated cardiomyocyte proliferation, and Pik3cb knockdown dampened the YAP mitogenic activity. Reciprocally, Yap loss-of-function impaired heart function and reduced cardiomyocyte proliferation and survival, all of which were significantly rescued by AAV-mediated Pik3cb expression.   Conclusion—Pik3cb is a crucial direct target of YAP, through which the YAP activates PI3K-AKT pathway and regulates cardiomyocyte proliferation and survival. Two groups were involved in this study:TNTcreYapfl_het group and TNTcreYapfl_KO group. Each group contained three biological replicates. Embryo hearts were collected at E12.5 and dissociated.  Cardiomyocytes were collected by FACS. The total RNA of cardiomyocytes were isolated for microarray analysis.							
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	Gu	Zhou	von Gise	Gu	Ma	Guo	van Gorp	Pu
Person First Name	Fei	Pingzhu	Alexander	Fei	Qing	Haidong	Pim	William
Person Mid Initials							R	T
Person Email	alickgf@hotmail.com							
Person Affiliation	Childrens hospital boston, Harvard Medical School							
Person Address	Cardiology, Childrens hospital boston, Harvard Medical School, 320 Longwood Ave., Boston, MA, USA							
Person Roles	submitter							
Protocol Name	P-GSE57718-1	P-GSE57718-5	P-GSE57718-6	P-GSE57718-2	P-GSE57718-3	P-GSE57718-4	P-GSE57718-7	
Protocol Description	The data were normalized using rma method ID_REF =  VALUE = RMA signal	Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 0.1ug total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).	Following fragmentation, the samples were hybridized on Mouse Gene 1.0 ST arrays at 45C with rotation set at 60 RPM for 16 hours using the GeneChip Hybridization Oven 640. The fluidics stations used for washing and staining are the Genechip Fluidics Station 450.	Mating of TNTCre, Yap flox and R26 mTmG mouse strains to yield Yap KO and Yap HET embryos with fluorescent marker, collection of embryos at E12.5, enzyme digestion of individual hearts and FACS for GFP labeled cardiomyocytes, collection in RLT buffer (Qiagen)	Mice were kept in a standard mouse room. All animal procedures were approved by the Institutional Animal Care and Use Committee.	Cell lysis in RLT buffer, followed by RNA purification on RNeasy mini columns according to the maunfacturers protocol	GeneChips were scanned using the Affymetrix GeneChip Scanner 3000 7G.	
Protocol Type	normalization data transformation protocol	labelling protocol	hybridization protocol	sample treatment protocol	growth protocol	nucleic acid extraction protocol	array scanning protocol	
Publication Title	Pi3kcb links Hippo-YAP and PI3K-AKT signaling pathways to promote cardiomyocyte proliferation and survival.							
Publication Author List	Lin Z, Zhou P, von Gise A, Gu F, Ma Q, Chen J, Guo H, van Gorp PR, Wang DZ, Pu WT							
PubMed ID	25249570							
Publication DOI	10.1161/CIRCRESAHA.115.304457							
Comment[SecondaryAccession]	GSE57718							
Comment[GEOReleaseDate]	2015-02-20							
Comment[ArrayExpressSubmissionDate]	2014-05-15							
Comment[GEOLastUpdateDate]	2015-02-20							
Comment[AEExperimentType]	transcription profiling by array							
SDRF File	E-GEOD-57718.sdrf.txt