title: Diphtheria toxin fused to granulocyte-macrophage colony-stimulating factor is toxic to blasts from patients with juvenile myelomonocytic leukemia and chronic myelomonocytic leukemia.
abstract: We have previously demonstrated that human granulocyte-macrophage colony-stimulating factor fused to a truncated diphtheria toxin (DT388-GM-CSF) is toxic to patient acute myeloid leukemia progenitors bearing the GM-CSF receptor, but not normal marrow progenitors. We now report that exposure of mononuclear cells from five of seven (71%) juvenile myelomonocytic leukemia (JMML) patients and from 12 of 20 (60%) adult chronic myelomonocytic leukemia (CMML) patients to 10(-9) mol/L DT388-GM-CSF for 48 hours in culture reduces the number of cells capable of forming colonies in semisolid medium (colony-forming units-leukemia) 10-fold to 300-fold (1 to 2.5 log decrease). In contrast, normal myeloid progenitors (colony-forming unit-granulocyte-macrophage) from six different donors treated and assayed under identical conditions were consistently insensitive to the same fusion toxin even when treated as highly purified CD34(+) cells. The leukemic progenitors from the two other JMML patients showed intermediate sensitivity to DT388-GM-CSF and the leukemic progenitors from eight of the 20 (40%) CMML patients were not different from normal progenitors. Parallel measurements of the number and affinity of GM-CSF receptors on cells from the same samples showed no consistent differences between JMML, CMML, and normal light density or CD34(+) bone marrow cells. The increased sensitivity of leukemic progenitors from all JMML progenitors and some CMML patients to the fusion toxin is therefore not likely to be explained by an increased density of GM-CSF receptors on these cells. We also examined the DT388-GM-CSF sensitivity of two murine cell lines transfected with cDNAs encoding varying portions of the human GM-CSF receptor and/or beta chains. These studies showed that high-affinity ligand binding was sufficient for DT388-GM-CSF-induced toxicity, as this could occur even in the absence of functional signal transduction and that the background of the host cell had a major influence on the degree to which this decreased the toxicity of DT388-GM-CSF. The selective sensitivity to DT388-GM-CSF of leukemic progenitors from a majority of JMML and CMML patients suggests that this agent could have therapeutic potential for some patients with these diseases.
authors.affiliation: Wake Forest Comprehensive Cancer Center/Bowman Gray School of Medicine, Winston-Salem, NC, USA.
authors.forename: A E
authors.initials: AE
authors.lastname: Frankel
authors.forename: M
authors.initials: M
authors.lastname: Lilly
authors.forename: R
authors.initials: R
authors.lastname: Kreitman
authors.forename: D
authors.initials: D
authors.lastname: Hogge
authors.forename: M
authors.initials: M
authors.lastname: Beran
authors.forename: M H
authors.initials: MH
authors.lastname: Freedman
authors.forename: P D
authors.initials: PD
authors.lastname: Emanuel
authors.forename: C
authors.initials: C
authors.lastname: McLain
authors.forename: P
authors.initials: P
authors.lastname: Hall
authors.forename: E
authors.initials: E
authors.lastname: Tagge
authors.forename: M
authors.initials: M
authors.lastname: Berger
authors.forename: C
authors.initials: C
authors.lastname: Eaves
mesh_terms: D000328:Adult; D000368:Aged; D016923:Cell Death / Q000187:drug effects; D002675:Child, Preschool; D004167:Diphtheria Toxin / Q000235:genetics / Q000494:pharmacology* / Q000627:therapeutic use; D005260:Female; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics / Q000494:pharmacology* / Q000627:therapeutic use; D006801:Humans; D015477:Leukemia, Myelomonocytic, Chronic / Q000188:drug therapy / Q000473:pathology*; D008232:Lymphoproliferative Disorders / Q000188:drug therapy / Q000473:pathology; D008297:Male; D008875:Middle Aged; D014411:Neoplastic Stem Cells / Q000187:drug effects* / Q000473:pathology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000494:pharmacology* / Q000627:therapeutic use
chemical_list: D004167:Diphtheria Toxin; D011993:Recombinant Fusion Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor