title: Prevalidation of a model for predicting acute neutropenia by colony forming unit granulocyte/macrophage (CFU-GM) assay.
abstract: This report describes an international prevalidation study conducted to optimise the Standard Operating Procedure (SOP) for detecting myelosuppressive agents by CFU-GM assay and to study a model for predicting (by means of this in vitro hematopoietic assay) the acute xenobiotic exposure levels that cause maximum tolerated decreases in absolute neutrophil counts (ANC). In the first phase of the study (Protocol Refinement), two SOPs were assessed, by using two cell culture media (Test A, containing GM-CSF; and Test B, containing G-CSF, GM-CSF, IL-3, IL-6 and SCF), and the two tests were applied to cells from both human (bone marrow and umbilical cord blood) and mouse (bone marrow) CFU-GM. In the second phase (Protocol Transfer), the SOPs were transferred to four laboratories to verify the linearity of the assay response and its interlaboratory reproducibility. After a further phase (Protocol Performance), dedicated to a training set of six anticancer drugs (adriamycin, flavopindol, morpholino-doxorubicin, pyrazoloacridine, taxol and topotecan), a model for predicting neutropenia was verified. Results showed that the assay is linear under SOP conditions, and that the in vitro endpoints used by the clinical prediction model of neutropenia are highly reproducible within and between laboratories. Valid tests represented 95% of all tests attempted. The 90% inhibitory concentration values (IC(90)) from Test A and Test B accurately predicted the human maximum tolerated dose (MTD) for five of six and for four of six myelosuppressive anticancer drugs, respectively, that were selected as prototype xenobiotics. As expected, both tests failed to accurately predict the human MTD of a drug that is a likely protoxicant. It is concluded that Test A offers significant cost advantages compared to Test B, without any loss of performance or predictive accuracy. On the basis of these results, we proposed a formal Phase II validation study using the Test A SOP for 16-18 additional xenobiotics that represent the spectrum of haematotoxic potential.
authors.affiliation: Institute of Microbiology, Via Pascal 36, 20133, Milan, Italy. pessinaa@mailserver.unimi.it
authors.forename: A
authors.initials: A
authors.lastname: Pessina
authors.forename: B
authors.initials: B
authors.lastname: Albella
authors.forename: J
authors.initials: J
authors.lastname: Bueren
authors.forename: P
authors.initials: P
authors.lastname: Brantom
authors.forename: S
authors.initials: S
authors.lastname: Casati
authors.forename: L
authors.initials: L
authors.lastname: Gribaldo
authors.forename: C
authors.initials: C
authors.lastname: Croera
authors.forename: G
authors.initials: G
authors.lastname: Gagliardi
authors.forename: P
authors.initials: P
authors.lastname: Foti
authors.forename: R
authors.initials: R
authors.lastname: Parchment
authors.forename: D
authors.initials: D
authors.lastname: Parent-Massin
authors.forename: Y
authors.initials: Y
authors.lastname: Sibiril
authors.forename: R
authors.initials: R
authors.lastname: Van Den Heuvel
mesh_terms: D000208:Acute Disease; D000818:Animals; D001854:Bone Marrow Cells; D002452:Cell Count; D003114:Colony-Forming Units Assay; D004285:Dogs; D005312:Fetal Blood; D006098:Granulocytes / Q000187:drug effects* / Q000473:pathology; D006801:Humans; D008264:Macrophages / Q000187:drug effects* / Q000473:pathology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D008954:Models, Biological; D009503:Neutropenia / Q000139:chemically induced* / Q000473:pathology; D011237:Predictive Value of Tests; D051381:Rats; D011916:Rats, Inbred F344; D015203:Reproducibility of Results; D015262:Xenobiotics / Q000633:toxicity*
chemical_list: D015262:Xenobiotics