title: In vitro to in vivo concordance of a high throughput assay of bone marrow toxicity across a diverse set of drug candidates.
abstract: The development of predictive toxicology assays is necessary to optimize the drug candidate selection process. The colony forming assay (CFA) is used routinely to assess bone marrow toxicity and represents a viable tool for the discovery toxicologist, but the assay is not widely accepted as a standard screening tool due to technical challenges. A higher throughput and standardized version of the assay recently was developed such that the proliferative capacity of a cell lineage is measured indirectly via ATP levels, replacing the cumbersome identification and enumeration of specific colonies. In this study, a high-throughput assay of bone marrow toxicity prediction using the granulocyte, erythrocyte, monocyte, and macrophage (GEMM) progenitor cell lineage was evaluated using a training set of 56 structurally diverse compounds with known in vivo bone marrow effects. In general, compounds identified as toxic in vivo had lower IC(50) values, whereas those identified as non-toxic had higher IC(50) values. Concordance (i.e., predictive accuracy) to in vivo bone marrow toxicity results was 82% when an in vitro toxicity threshold of 20 microM was used. Additional experiments in other hematopoietic lineages were conducted to determine if predictivity of several false positive and negative compounds in the GEMM lineage could be improved; however an increase in sensitivity or specificity was not observed. The high-throughput GEMM assay has good concordance to in vivo bone marrow toxicity results and, with the high-throughput and standardized format, can be incorporated readily into the pharmaceutical toxicological screening paradigm, aiding in the early identification of compounds that eventually may fail due to bone marrow toxicity.
authors.affiliation: Discovery and Investigative Safety, Non Clinical Safety, Roche Palo Alto, 3431 Hillview, Palo Alto, CA 94304, USA. andrew.olaharski@roche.com
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Olaharski
authors.forename: Hirdesh
authors.initials: H
authors.lastname: Uppal
authors.forename: Matthew
authors.initials: M
authors.lastname: Cooper
authors.forename: Stefan
authors.initials: S
authors.lastname: Platz
authors.forename: Tanja S
authors.initials: TS
authors.lastname: Zabka
authors.forename: Kyle L
authors.initials: KL
authors.lastname: Kolaja
mesh_terms: D000818:Animals; D001853:Bone Marrow / Q000187:drug effects*; D001854:Bone Marrow Cells / Q000187:drug effects*; D019070:Cell Lineage; D003114:Colony-Forming Units Assay / Q000379:methods*; D064420:Drug-Related Side Effects and Adverse Reactions; D006801:Humans; D066298:In Vitro Techniques; D004364:Pharmaceutical Preparations / Q000737:chemistry; D015203:Reproducibility of Results; D018675:Toxicity Tests / Q000379:methods*
chemical_list: D004364:Pharmaceutical Preparations