title: Quantitative real-time RT-PCR detects elevated Wilms tumor gene (WT1) expression in autologous blood stem cell preparations (PBSCs) from acute myeloid leukemia (AML) patients indicating contamination with leukemic blasts.
abstract: High-dose chemotherapy with subsequent autologous stem cell transplantation is believed to be of therapeutic benefit in patients with acute myeloid leukemia (AML), especially when no allogeneic bone marrow donor is available. One of the main risks is contamination of the stem cell preparations with leukemic blasts, which may account for a higher relapse rate compared to allogeneic bone marrow transplantation. Since overexpression of WT1 is common in leukemic blasts, we investigated, whether PBSCs from AML patients express WT1 at a higher level as compared to patients with solid cancers. PBSCs of seven patients with AML and of five patients with solid cancers were investigated for WT1 expression. Total WT1 copy count was determined in a standardized quantitative real time RT-PCR. WT1 expression was found in all AML PBSCs with an average copy number of 49.99 +/- 61.09. In solid cancers WT1 expression was statistically significantly lower with a copy number of 3.51 +/- 1.92. In AML patients with sustained complete remission we found a nearly significantly lower WT1 expression than in patients who relapsed within the first year after stem cell transplantation. Our data show a higher WT1 expression in PBSCs of AML patients compared to patients with solid cancers. This finding might indicate a contamination with leukemic blasts. Quantification of WT1 in PBSCs might therefore be useful to estimate the risk of relapse after autologous stem cell transplantation in AML patients.
authors.affiliation: Med. Klinik III (Hematology, Oncology and Transfusion Medicine), Universitätsklinikum Benjamin Franklin, Freie Universität Berlin, Berlin, Germany, EU.
authors.forename: J M
authors.initials: JM
authors.lastname: Siehl
authors.forename: E
authors.initials: E
authors.lastname: Thiel
authors.forename: R
authors.initials: R
authors.lastname: Leben
authors.forename: M
authors.initials: M
authors.lastname: Reinwald
authors.forename: W
authors.initials: W
authors.lastname: Knauf
authors.forename: H D
authors.initials: HD
authors.lastname: Menssen
mesh_terms: D000208:Acute Disease; D000293:Adolescent; D000328:Adult; D014408:Biomarkers, Tumor / Q000032:analysis; D005260:Female; D016162:Genes, Wilms Tumor / Q000502:physiology*; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D006801:Humans; D018189:Immunomagnetic Separation; D007937:Leukapheresis / Q000379:methods / Q000592:standards; D007951:Leukemia, Myeloid / Q000097:blood / Q000175:diagnosis / Q000628:therapy*; D008297:Male; D008875:Middle Aged; D011379:Prognosis; D012333:RNA, Messenger / Q000032:analysis; D012008:Recurrence; D012074:Remission Induction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D033581:Stem Cell Transplantation / Q000592:standards*; D014182:Transplantation, Autologous
chemical_list: D014408:Biomarkers, Tumor; D012333:RNA, Messenger