title: Multidrug resistance-associated protein 4 (MRP4) controls ganciclovir intracellular accumulation and contributes to ganciclovir-induced neutropenia in renal transplant patients.
abstract: Ganciclovir (GCV) is the cornerstone of cytomegalovirus prevention and treatment in transplant patients. It is associated with problematic adverse hematological effects in this population of immunosuppressed patients, which may lead to dose reduction thus favoring resistance. GCV crosses the membranes of cells, is activated by phosphorylation, and then stops the replication of viral DNA. Its intracellular accumulation might favor host DNA polymerase inhibition, hence toxicity. Following this hypothesis, we investigated the association between a selected panel of membrane transporter polymorphisms and the evolution of neutrophil counts in n=174 renal transplant recipients. An independent population of n=96 renal transplants served as a replication and experiments using HEK293T-transfected cells were performed to validate the clinical findings. In both cohorts, we found a variant in ABCC4 (rs11568658) associated with decreased neutrophil counts following valganciclovir (GCV prodrug) administration (exploratory cohort: β±SD=-0.68±0.28, p=0.029; replication cohort: β±SD=-0.84±0.29, p=0.0078). MRP4-expressing cells showed decreased GCV accumulation as compared to negative control cells (transfected with an empty vector) (-61%; p<0.0001). The efflux process was almost abolished in cells expressing MRP4 rs11568658 variant protein. Molecular dynamic simulations of GCV membrane crossing showed a preferred location of the drug just beneath the polar head group region, which supports its interaction with efflux transporters.
authors.affiliation: Univ. Limoges, UMR 850, F-87000 Limoges, France; INSERM, UMR 850, F-87000 Limoges, France.
authors.forename: Pierre-André
authors.initials: PA
authors.lastname: Billat
authors.affiliation: Univ. Limoges, UMR 850, F-87000 Limoges, France; INSERM, UMR 850, F-87000 Limoges, France.
authors.forename: Tahani
authors.initials: T
authors.lastname: Ossman
authors.affiliation: Univ. Limoges, UMR 850, F-87000 Limoges, France; INSERM, UMR 850, F-87000 Limoges, France; CHU Limoges, Service de Pharmacologie, Toxicologie et Pharmacovigilance, F-87000 Limoges, France.
authors.forename: Franck
authors.initials: F
authors.lastname: Saint-Marcoux
authors.affiliation: Univ. Limoges, UMR 850, F-87000 Limoges, France; INSERM, UMR 850, F-87000 Limoges, France; CHU Limoges, Service de Néphrologie, Dialyse et Transplantation, F-87000 Limoges, France.
authors.forename: Marie
authors.initials: M
authors.lastname: Essig
authors.affiliation: INSERM, UMR 850, F-87000 Limoges, France; CHU Limoges, Service de Néphrologie, Dialyse et Transplantation, F-87000 Limoges, France.
authors.forename: Jean-Philippe
authors.initials: JP
authors.lastname: Rerolle
authors.affiliation: CHU Toulouse Rangueil, Service de néphrologie, Dialyse et transplantation d⿿Organes, F-31000 Toulouse, France; INSERM, U1043, IFR⿿BMT, CHU Purpan, F-31000 Toulouse, France; Univ. Toulouse Paul Sabatier, F-31000 Toulouse, France.
authors.forename: Nassim
authors.initials: N
authors.lastname: Kamar
authors.affiliation: CHU Toulouse Rangueil, Service de néphrologie, Dialyse et transplantation d⿿Organes, F-31000 Toulouse, France; INSERM, U1043, IFR⿿BMT, CHU Purpan, F-31000 Toulouse, France; Univ. Toulouse Paul Sabatier, F-31000 Toulouse, France.
authors.forename: Lionel
authors.initials: L
authors.lastname: Rostaing
authors.affiliation: CHU Bordeaux, Service de Néphrologie, Transplantation, Dialyse, F-33000 Bordeaux, France.
authors.forename: Hannah
authors.initials: H
authors.lastname: Kaminski
authors.affiliation: Univ. Limoges, LCSN, Faculté de Pharmacie, F-87025 Limoges, France.
authors.forename: Gabin
authors.initials: G
authors.lastname: Fabre
authors.affiliation: Regional Centre of Advanced Technologies and Materials, Department of Physical Chemistry, Faculty of Science, Palacký University of Olomouc, Olomouc, Czech Republic.
authors.forename: Michal
authors.initials: M
authors.lastname: Otyepka
authors.affiliation: Univ. Limoges, UMR 850, F-87000 Limoges, France; INSERM, UMR 850, F-87000 Limoges, France; CHU Limoges, Service de Pharmacologie, Toxicologie et Pharmacovigilance, F-87000 Limoges, France.
authors.forename: Jean-Baptiste
authors.initials: JB
authors.lastname: Woillard
authors.affiliation: Univ. Limoges, UMR 850, F-87000 Limoges, France; INSERM, UMR 850, F-87000 Limoges, France; CHU Limoges, Service de Pharmacologie, Toxicologie et Pharmacovigilance, F-87000 Limoges, France.
authors.forename: Pierre
authors.initials: P
authors.lastname: Marquet
authors.affiliation: Univ. Limoges, UMR 850, F-87000 Limoges, France; INSERM, UMR 850, F-87000 Limoges, France; Regional Centre of Advanced Technologies and Materials, Department of Physical Chemistry, Faculty of Science, Palacký University of Olomouc, Olomouc, Czech Republic.
authors.forename: Patrick
authors.initials: P
authors.lastname: Trouillas
authors.affiliation: Univ. Limoges, UMR 850, F-87000 Limoges, France; INSERM, UMR 850, F-87000 Limoges, France; CHU Limoges, Service de Pharmacologie, Toxicologie et Pharmacovigilance, F-87000 Limoges, France. Electronic address: nicolas.picard@unilim.fr.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Picard
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000998:Antiviral Agents / Q000009:adverse effects / Q000493:pharmacokinetics / Q000627:therapeutic use; D003586:Cytomegalovirus Infections / Q000517:prevention & control; D005260:Female; D015774:Ganciclovir / Q000009:adverse effects / Q000493:pharmacokinetics; D057809:HEK293 Cells; D006801:Humans; D019169:Jurkat Cells; D016030:Kidney Transplantation; D008297:Male; D008875:Middle Aged; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000235:genetics / Q000378:metabolism*; D009503:Neutropenia / Q000139:chemically induced* / Q000235:genetics / Q000378:metabolism; D020641:Polymorphism, Single Nucleotide; D055815:Young Adult
chemical_list: D000998:Antiviral Agents; D015774:Ganciclovir; D027425:ATP-Binding Cassette, Sub-Family C Proteins; C073492:ABCC4 protein, human
keywords: Cytomegalovirus; Ganciclovir; Membrane transporters; Multidrug resistance-associated protein 4; Pharmacogenetics; Transplantation