title: Expression of the Wilms' tumor gene (WT1) in normal hemopoiesis.
abstract: The Wilms tumor suppressor gene (WT1) is mutated in a number of cases of Wilms' tumor as well as in mesothelioma and leukemia. It encodes a transcription factor derived from any one of four alternate transcripts. WT1 has a restricted pattern of expression within the body and within the hemopoietic system its expression is limited to primitive leukemias and a number of leukemic cell lines. Given the overexpression of WT1 in leukemias, we have addressed the question of whether this gene is expressed within the normal hemopoietic system. Mononuclear bone marrow (BM) cells obtained from normal donors were separated by fluorescence-activated cell sorting (FACS) into "primitive" (CD34+) and "mature" (CD34-) cell populations. Total RNA extracted from these cells was subjected to reverse transcriptase polymerase chain reaction (RT-PCR) using primers based on the WT1 sequence, to examine the expression of this gene within the hemopoietic system. Phenotypic purity of cells was guaranteed by performing single-cell sorting followed by RT-PCR to define the precise cellular phenotypes that express WT1. Expression of WT1 was detected in cells bearing the CD34+ phenotype but not in those cells lacking expression of CD34. In addition, single-cell analysis revealed that expression of WT1 occurred in the candidate stem cell-containing population of hemopoietic cells which have the phenotype CD34+ CD38-. Moreover, the single-cell RT-PCR analysis also demonstrated that differential expression of alternate transcripts of WT1 occurs between hemopoietic progenitor cells with the same phenotype. In conclusion, expression of WT1 is limited to early progenitors of the blood system, which suggests that this gene plays a critical role in hemopoietic development.
authors.affiliation: Leukaemia Research Unit, Hanson Centre for Cancer Research, Adelaide, South Australia, Australia.
authors.forename: P N
authors.initials: PN
authors.lastname: Baird
authors.forename: P J
authors.initials: PJ
authors.lastname: Simmons
mesh_terms: D018952:Antigens, CD34; D001854:Bone Marrow Cells / Q000166:cytology; D004268:DNA-Binding Proteins / Q000235:genetics*; D015870:Gene Expression; D006410:Hematopoiesis / Q000235:genetics*; D006412:Hematopoietic Stem Cells / Q000166:cytology; D006801:Humans; D014157:Transcription Factors / Q000235:genetics*; D025721:WT1 Proteins
chemical_list: D018952:Antigens, CD34; D004268:DNA-Binding Proteins; D014157:Transcription Factors; D025721:WT1 Proteins