title: A cell-based approach for the early assessment of the phospholipidogenic potential in pharmaceutical research and drug development.
abstract: Phospholipidosis is a term commonly used to indicate a phospholipid storage disorder; in affected cells, phospholipids accumulate in lysosomes that acquire a multilamellar morphological appearance. Cationic amphiphilic drugs (CADs) are suggested to induce phospholipidosis by direct interaction of xenobiotics with intracellular phospholipids or by the action of xenobiotics on the synthesis and metabolism of phospholipids. To date, electron microscopy (EM) represents the most reliable and the preferred method for the demonstration of phospholipidotic cell damage. Nevertheless, EM has a low throughput, it is expensive, and it is not suitable for screening purposes. We discuss here the assessment of the the phospholipidogenic potential of drugs using a cell culture-based model. In this test, intracellular phospholipids of treated U-937 cells (a human monocyte-derived cell line) were measured using the fluorescent probe Nile red. Eleven CADs reported to induce phospholipidosis in vivo and eight nonphospholipidogenic drugs were tested. Results obtained with the U-937 model confirmed the phospholipidogenic potential of drugs tested as described in the literature. Results have also been correlated with data obtained with a physical-chemical model (chromatographic hydrophobicity index measurement). Good correlation was obtained, confirming that the physical-chemical properties of CADs play a crucial role in the development of phospholipidosis. This work demonstrates that the U-937 model is a rapid and sensitive method for the determination of phospholipidosis-mediated cell damage. The specificity and the predictive potency observed make this method suitable for screening purposes in pharmaceutical development.
authors.affiliation: Safety Assessment Department, GlaxoSmithKline Research Centre, Verona, Italy. AC48574@gsk.com
authors.forename: A
authors.initials: A
authors.lastname: Casartelli
authors.forename: M
authors.initials: M
authors.lastname: Bonato
authors.forename: P
authors.initials: P
authors.lastname: Cristofori
authors.forename: F
authors.initials: F
authors.lastname: Crivellente
authors.forename: G
authors.initials: G
authors.lastname: Dal Negro
authors.forename: I
authors.initials: I
authors.lastname: Masotto
authors.forename: C
authors.initials: C
authors.lastname: Mutinelli
authors.forename: K
authors.initials: K
authors.lastname: Valko
authors.forename: V
authors.initials: V
authors.lastname: Bonfante
mesh_terms: D002412:Cations; D002470:Cell Survival; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D005434:Flow Cytometry; D006801:Humans; D008064:Lipidoses / Q000139:chemically induced; D010078:Oxazines / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism*; D020127:Recovery of Function; D013194:Staining and Labeling / Q000379:methods; D013501:Surface-Active Agents / Q000633:toxicity*; D020298:U937 Cells / Q000187:drug effects / Q000378:metabolism*; D015262:Xenobiotics / Q000145:classification / Q000633:toxicity*
chemical_list: D002412:Cations; D010078:Oxazines; D010743:Phospholipids; D013501:Surface-Active Agents; D015262:Xenobiotics; C044808:nile red