title: Functional screen identifies regulators of murine hematopoietic stem cell repopulation.
abstract: Understanding the molecular regulation of hematopoietic stem and progenitor cell (HSPC) engraftment is paramount to improving transplant outcomes. To discover novel regulators of HSPC repopulation, we transplanted >1,300 mice with shRNA-transduced HSPCs within 24 h of isolation and transduction to focus on detecting genes regulating repopulation. We identified 17 regulators of HSPC repopulation: Arhgef5, Armcx1, Cadps2, Crispld1, Emcn, Foxa3, Fstl1, Glis2, Gprasp2, Gpr56, Myct1, Nbea, P2ry14, Smarca2, Sox4, Stat4, and Zfp251. Knockdown of each of these genes yielded a loss of function, except in the cases of Armcx1 and Gprasp2, whose loss enhanced hematopoietic stem cell (HSC) repopulation. The discovery of multiple genes regulating vesicular trafficking, cell surface receptor turnover, and secretion of extracellular matrix components suggests active cross talk between HSCs and the niche and that HSCs may actively condition the niche to promote engraftment. We validated that Foxa3 is required for HSC repopulating activity, as Foxa3(-/-) HSC fails to repopulate ablated hosts efficiently, implicating for the first time Foxa genes as regulators of HSPCs. We further show that Foxa3 likely regulates the HSC response to hematologic stress. Each gene discovered here offers a window into the novel processes that regulate stable HSPC engraftment into an ablated host.
authors.affiliation: Department of Hematology, St. Jude Children's Research Hospital, Memphis, TN 38105.
authors.forename: Per
authors.initials: P
authors.lastname: Holmfeldt
authors.affiliation: Department of Hematology, St. Jude Children's Research Hospital, Memphis, TN 38105.
authors.forename: Miguel
authors.initials: M
authors.lastname: Ganuza
authors.affiliation: Department of Hematology, St. Jude Children's Research Hospital, Memphis, TN 38105.
authors.forename: Himangi
authors.initials: H
authors.lastname: Marathe
authors.affiliation: Interdisciplinary Graduate Program in Genetics, University of Iowa, Iowa City, IA 52242.
authors.forename: Bing
authors.initials: B
authors.lastname: He
authors.affiliation: Department of Hematology, St. Jude Children's Research Hospital, Memphis, TN 38105.
authors.forename: Trent
authors.initials: T
authors.lastname: Hall
authors.affiliation: Department of Biostatistics, St. Jude Children's Research Hospital, Memphis, TN 38105.
authors.forename: Guolian
authors.initials: G
authors.lastname: Kang
authors.affiliation: Department of Biostatistics, St. Jude Children's Research Hospital, Memphis, TN 38105.
authors.forename: Joseph
authors.initials: J
authors.lastname: Moen
authors.affiliation: Department of Hematology, St. Jude Children's Research Hospital, Memphis, TN 38105.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Pardieck
authors.affiliation: Department of Hematology, St. Jude Children's Research Hospital, Memphis, TN 38105.
authors.forename: Angelica C
authors.initials: AC
authors.lastname: Saulsberry
authors.affiliation: Department of Hematology, St. Jude Children's Research Hospital, Memphis, TN 38105.
authors.forename: Alba
authors.initials: A
authors.lastname: Cico
authors.affiliation: Department of Hematology, St. Jude Children's Research Hospital, Memphis, TN 38105.
authors.forename: Ludovic
authors.initials: L
authors.lastname: Gaut
authors.affiliation: Department of Computational Biology, St. Jude Children's Research Hospital, Memphis, TN 38105.
authors.forename: Daniel
authors.initials: D
authors.lastname: McGoldrick
authors.affiliation: Department of Computational Biology, St. Jude Children's Research Hospital, Memphis, TN 38105.
authors.forename: David
authors.initials: D
authors.lastname: Finkelstein
authors.affiliation: Department of Internal Medicine, University of Iowa, Iowa City, IA 52242.
authors.forename: Kai
authors.initials: K
authors.lastname: Tan
authors.affiliation: Department of Hematology, St. Jude Children's Research Hospital, Memphis, TN 38105 shannon.mckinney-freeman@stjude.org.
authors.forename: Shannon
authors.initials: S
authors.lastname: McKinney-Freeman
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D049109:Cell Proliferation; D019610:Cytoprotection; D004742:Enhancer Elements, Genetic / Q000235:genetics; D056726:Genetic Association Studies; D005820:Genetic Testing; D006410:Hematopoiesis; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D051543:Hepatocyte Nuclear Factor 3-gamma / Q000378:metabolism; D008810:Mice, Inbred C57BL; D011485:Protein Binding; D015203:Reproducibility of Results; D015398:Signal Transduction; D013312:Stress, Physiological
chemical_list: C496506:Foxa3 protein, mouse; D051543:Hepatocyte Nuclear Factor 3-gamma