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Update on StemBase, the database of the Stem Cell Genomics Project Miguel Andrade, OHRI, Ottawa Stem Cell Genomics Project StemBase DVD release Oct 2005 Stem Cell Genomics Project StemBase DVD release Oct 2005 The Stem Cell Genomics Project • Objective: acquire a complete understanding of the genetic factors that: – specify stem cell identity and function; and – regulate commitment and differentiation • Rationale: – Stem cells play an essential role in the human body as they provide the starting material for every organ and tissue – Knowledge of regulatory genes acting in and on stem cells is necessary to exploit their full therapeutic potential STEM CELL NETWORK +20 GROUPS samples OGIC – GENOMICS PLATFORM Pearl Campbell data Bioinformatics Group data StemBase PUBLIC Yauk CL, Berndt ML, Williams A, Douglas GR. 2004. Comprehensive comparison of six microarray technologies. Nucleic Acids Research. 32(15): p. e124 Data sharing model 4 Months each sample Submitter 1 Sample Data production 3 Data 2 Data NO 12 months entire StemBase 4 Approve release? 5 Embargo or remove YES Bioinformatics analysis Working StemBase 5 Publish data Published StemBase Network investigators 6 Publicize data Public StemBase SCN Sample Contributors • • • • • • • • • • • • • • • Jane Aubin Mickie Bhatia John Dick Connie Eaves Jaques Galipeau Alain Garnier Marina Gertsenstein May Griffith John Hassell Norman Iscove Michael McBurney Rod McInnes Kelly McNagny Lynn Megeney James Piret • • • • • • • • • • • • • • Derrick Rancourt Janet Rossant Lawrence Rosenberg Michael Rudnicki Luc Sabourin Guy Savageau Ruth Slack Jacques P. Tremblay Valerie Wallace T. Michael Underhill Sue Varmuza Samuel Weiss Juan Carlos Zuniga-Pflucker Peter Zandstra Data collection 191 samples Retina Neural Myoblasts Muscle 1/4 Human 3/4 Mouse … Rat Adipose Tissue Blastocyst Blood Mammary gland Bone marrow Limb bud Larynx Kidney Fibroblasts Fetal liver Embryonic Carcinoma Brain Calvaria Cervix epithelium Embryonic Ciliary margin of eye Cord Blood Dermis Stem Cell Genomics Project StemBase DVD release 1 Oct 2005 StemBase Database of gene expression data in mouse and human stem cells Study genes important for stem cell function Affymetrix DNA microarray data. 185 samples. Serial Analysis of Gene Expression. 6 samples. Perez-Iratxeta, C., G. Palidwor, C.J. Porter, N.A. Sanche, M.R. Huska, B.P. Suomela, E.M. Muro, P. Krzyzanowski, E. Hughes, P.A. Campbell, M.A. Rudnicki and M.A. Andrade. 2005. Study of stem cell function using microarray experiments. FEBS Letters. 579, 1795-1801. Usage (1 Nov 2005) 258 non SCN accounts 3,000 logins First commercial license Public web server http://www.scgp.ca:8080/StemBase/ Data submission to GEO Data submission to GEO Required by some journals Largest gene expression data repository StemBase: High quality data Focused on stem cells More functionality GEO as a traffic catch Mouse / MOE430 V6.5 R1 D4A Embyoid bodies C2D C2A C2E J1 Embyoid bodies D4E Myoblasts Cancer Cancer R1 serum64 R1 serum6999 D4D Embyoid bodies Cancer Dim2 Neurospheres Retinal first passage Dermis Adipose Retinal primary Mammospheres Osteoblasts Neural Bone marrow Bone marrow Myospheres Dim1 Bone marrow Mammospheres undifferenciated Muscle Bone marrow Carolina Perez-Iratxeta Human / HGU133 Cord blood Bone marrow M-O7e Smad7 Dim2 Cord blood M-O7e Peripheral M-O7e Fetal Peripheral Hela I6 Cord blood Bone marrow Kidney Hela Retinal primary Myoblasts Retinal first passage Dim1 I6 Myoblasts differentiated Carolina Perez-Iratxeta Analysis of genes correlated with Oct4 ES EB self-renewal totipotency lineage commitment Oct4 Hypothesis: Analysis of genes correlated with Oct4 in stem cells will allow the identification of genes that are important for stem cell identity and lineage commitment Pearl Campbell Paul Krzyzanowski Time Series Time series analysis: differentiation three types of mESC (R1, J1, V6.5) into embryoid bodies 11 time points: 0hr, 6hr, 12hr, 18hr, 24hr, 36hr, 48h, 4day, 7day, 9day and 14day Hailesellasse et al. 2005. Search for genes important in mouse embryonic stem cell differentiation. Submitted. 6h<0h 12h<0h 12h>0h 6h>0h P1 Egr1 P2 Klf4 P3 Klf4 P4 D930005D10Rik P5 Pim3 P6 Rtn4 P7 - P8 Socs3 P9 Fblim1 P10 Epha5 P11 Sfrs5 P12 Tagln P13 1190003J15Rik P14 Herc1 P15 Spry4 P16 Fblim1 P17 Mras P18 Anxa3 P19 Zfp54 P20 Rbm5 P21 4930431H11Rik P22 Khsrp P23 1110061A14Rik P24 Rbm14 b. 1190003J15Rik a. Fblim1 from down-regulated genes / signaling related c. Socs3 TR_THY SOCS box d. Epha5 (Eph receptor A5) f. Tagln e. Pim3 Calponin repeat SAM 2 g. Herc1 RCC1 i. Mras h. Spry4 WD40 SPRY h. Anxa3 RCC1 HECTc j. Rtn4 l. D930005D10Rik Annexin repeats Zn finger domains m. Klf4 n. Egr1 from down-regulated genes Zn finger domains o. Sfrs5 / nucleotide binding RRM_1 RRM_1 from up-regulated genes q. Rbm5 p. Rbm14 RRM_1 zf-RanBP s. Khsrp r. Zfp54 Zn finger domains G-patch 1190003J15Rik Signal transduction (GO:0007165) Fblim1 Spry4 Anxa3 Morphogenesis (GO:000653) Zinc ion binding (GO:0043167) DNA binding D930005D10Rik Rtn4 Tagln Protein amino acid phosphorylation (GO:0006468) Socs3 Egr1 Zfp54 Mras Rbm14 Epha5 Pim3 Herc1 Klf4 Rbm5 Nucleic acid binding (GO:0003676) Sfrs5 Khsrp RNA binding Protein modification (GO:0006464) Future Integration of proteomics data New analysis features External datasets: More stem cell data Cancer sets (e.g. NCI) Normal tissue Stem Cell Genomics Project StemBase DVD release Oct 2005 DVD release Oct 2005 Raw microarray data in StemBase as of 23 Oct 2005 154 samples 426 replicates 4 DVDs Lily Jin / Gareth Palidwor Ottawa Genome Centre Michael Rudnicki (director) William Read (project manager) Proteomics Lawrence Puente Lynn Megeney Genomics Core Facility Pearl Campbell Stem Cell Network Norman Iscove (U of T) Tim Hughes (U of T) Bioinformatics Enrique Muro Gareth Palidwor Carolina Perez-Iratxeta Neal Sanche (03-04) Lily Jin (-Sept05) Miguel Andrade Kagnew Hailesellasse (-Aug05) Christopher Porter Paul Krzyzanowski Andrew Kysyk Thanks! http://www.scgp.ca:8080/StemBase http://www.ottawagenomecenter.ca