title: The Wilms' tumor gene WT1-GFP knock-in mouse reveals the dynamic regulation of WT1 expression in normal and leukemic hematopoiesis.
abstract: The Wilms' tumor gene WT1 is overexpressed in most of human leukemias regardless of disease subtypes. To characterize the expression pattern of WT1 during normal and neoplastic hematopoiesis, we generated a knock-in reporter green fluorescent protein (GFP) mouse (WT1(GFP/+)) and assayed for WT1 expression in normal and leukemic hematopoietic cells. In normal hematopoietic cells, WT1 was expressed in none of the long-term (LT) hematopoietic stem cells (HSC) and very few (<1%) of the multipotent progenitor cells. In contrast, in murine leukemias induced by acute myeloid leukemia 1 (AML1)/ETO+TEL/PDGFbetaR or BCR/ABL, WT1 was expressed in 40.5 or 38.9% of immature c-kit(+)lin(-)Sca-1(+) (KLS) cells, which contained a subset, but not all, of transplantable leukemic stem cells (LSCs). WT1 expression was minimal in normal fetal liver HSCs and mobilized HSCs, both of which are stimulated for proliferation. In addition, overexpression of WT1 in HSCs did not result in proliferation or expansion of HSCs and their progeny in vivo. Thus, the mechanism by which expansion of WT1-expressing cells occurs in leukemia remains unclear. Nevertheless, our results demonstrate that the WT1(GFP/+) mouse is a powerful tool for analyzing WT1-expressing cells, and they highlight the potential of WT1, as a specific therapeutic target that is expressed in LSCs but not in normal HSCs.
authors.affiliation: Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA. hnaoki@stanford.edu
authors.forename: N
authors.initials: N
authors.lastname: Hosen
authors.forename: T
authors.initials: T
authors.lastname: Shirakata
authors.forename: S
authors.initials: S
authors.lastname: Nishida
authors.forename: M
authors.initials: M
authors.lastname: Yanagihara
authors.forename: A
authors.initials: A
authors.lastname: Tsuboi
authors.forename: M
authors.initials: M
authors.lastname: Kawakami
authors.forename: Y
authors.initials: Y
authors.lastname: Oji
authors.forename: Y
authors.initials: Y
authors.lastname: Oka
authors.forename: M
authors.initials: M
authors.lastname: Okabe
authors.forename: B
authors.initials: B
authors.lastname: Tan
authors.forename: H
authors.initials: H
authors.lastname: Sugiyama
authors.forename: I L
authors.initials: IL
authors.lastname: Weissman
mesh_terms: D000818:Animals; D001853:Bone Marrow; D049109:Cell Proliferation; D003114:Colony-Forming Units Assay; D004195:Disease Models, Animal; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D016162:Genes, Wilms Tumor; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D016130:Immunophenotyping; D016086:Lentivirus; D007942:Leukemia, Experimental / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D014411:Neoplastic Stem Cells / Q000473:pathology; D014162:Transfection; D025721:WT1 Proteins / Q000235:genetics / Q000502:physiology*
chemical_list: D025721:WT1 Proteins; D049452:Green Fluorescent Proteins