title: A loss of function screen of identified genome-wide association study Loci reveals new genes controlling hematopoiesis.
abstract: The formation of mature cells by blood stem cells is very well understood at the cellular level and we know many of the key transcription factors that control fate decisions. However, many upstream signalling and downstream effector processes are only partially understood. Genome wide association studies (GWAS) have been particularly useful in providing new directions to dissect these pathways. A GWAS meta-analysis identified 68 genetic loci controlling platelet size and number. Only a quarter of those genes, however, are known regulators of hematopoiesis. To determine function of the remaining genes we performed a medium-throughput genetic screen in zebrafish using antisense morpholino oligonucleotides (MOs) to knock down protein expression, followed by histological analysis of selected genes using a wide panel of different hematopoietic markers. The information generated by the initial knockdown was used to profile phenotypes and to position candidate genes hierarchically in hematopoiesis. Further analysis of brd3a revealed its essential role in differentiation but not maintenance and survival of thrombocytes. Using the from-GWAS-to-function strategy we have not only identified a series of genes that represent novel regulators of thrombopoiesis and hematopoiesis, but this work also represents, to our knowledge, the first example of a functional genetic screening strategy that is a critical step toward obtaining biologically relevant functional data from GWA study for blood cell traits.
authors.affiliation: Department of Haematology, University of Cambridge, Cambridge, United Kingdom; Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, United Kingdom; NHS Blood and Transplant, Cambridge, United Kingdom.
authors.forename: Ewa
authors.initials: E
authors.lastname: Bielczyk-Maczyńska
authors.affiliation: MRC Centre for Developmental and Biomedical Genetics, University of Sheffield, Sheffield, United Kingdom; Department of Cardiovascular Sciences, University of Sheffield, Sheffield, United Kingdom.
authors.forename: Jovana
authors.initials: J
authors.lastname: Serbanovic-Canic
authors.affiliation: Department of Haematology, University of Cambridge, Cambridge, United Kingdom; Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, United Kingdom.
authors.forename: Lauren
authors.initials: L
authors.lastname: Ferreira
authors.affiliation: Department of Haematology, University of Cambridge, Cambridge, United Kingdom; Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, United Kingdom.
authors.forename: Nicole
authors.initials: N
authors.lastname: Soranzo
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, United Kingdom.
authors.forename: Derek L
authors.initials: DL
authors.lastname: Stemple
authors.affiliation: Department of Haematology, University of Cambridge, Cambridge, United Kingdom; Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, United Kingdom; NHS Blood and Transplant, Cambridge, United Kingdom.
authors.forename: Willem H
authors.initials: WH
authors.lastname: Ouwehand
authors.affiliation: Department of Haematology, University of Cambridge, Cambridge, United Kingdom; Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, United Kingdom.
authors.forename: Ana
authors.initials: A
authors.lastname: Cvejic
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000235:genetics*; D020869:Gene Expression Profiling; D056426:Genetic Loci; D055106:Genome-Wide Association Study; D006410:Hematopoiesis / Q000235:genetics*; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D020641:Polymorphism, Single Nucleotide; D015027:Zebrafish / Q000097:blood