Proteomics of human liver membrane transporters: a focus on fetuses and newborn infants

• Published in: European journal of pharmaceutical sciences 2018-11, Vol.124, p.217-227
• Main Authors: van Groen, Bianca D., van de Steeg, Evita, Mooij, Miriam G., van Lipzig, Marola M.H., de Koning, Barbara A.E., Verdijk, Robert M., Wortelboer, Heleen M., Gaedigk, Roger, Bi, Chengpeng, Leeder, J. Steven, van Schaik, Ron H.N., van Rosmalen, Joost, Tibboel, Dick, Vaes, Wouter H., de Wildt, Saskia N.
• Format: Article
• Language: English
• Subjects: Adult; Animals; Child; Child, Preschool; Dogs; Fetus - metabolism; HEK293 Cells; Humans; Infant; Infant, Newborn; Liver; Liver - embryology; Liver - metabolism; Madin Darby Canine Kidney Cells; Membrane Transport Proteins - genetics; Membrane Transport Proteins - metabolism; Ontogeny; Pediatrics; Proteomics; RNA, Messenger - metabolism; Transporters
• ISSN: 0928-0987; 1879-0720
• DOI: 10.1016/j.ejps.2018.08.042

Hepatic membrane transporters are involved in the transport of many endogenous and exogenous compounds, including drugs. We aimed to study the relation of age with absolute transporter protein expression in a cohort of 62 mainly fetus and newborn samples. Protein expressions of BCRP, BSEP, GLUT1, MCT1, MDR1, MRP1, MRP2, MRP3, NTCP, OCT1, OATP1B1, OATP1B3, OATP2B1 and ATP1A1 were quantified with LC-MS/MS in isolated crude membrane fractions of snap-frozen post-mortem fetal and pediatric, and surgical adult liver samples. mRNA expression was quantified using RNA sequencing, and genetic variants with TaqMan assays. We explored relationships between protein expression and age (gestational age [GA], postnatal age [PNA], and postmenstrual age); between protein and mRNA expression; and between protein expression and genotype. We analyzed 36 fetal (median GA 23.4 weeks [range 15.3–41.3]), 12 premature newborn (GA 30.2 weeks [24.9–36.7], PNA 1.0 weeks [0.14–11.4]), 10 term newborn (GA 40.0 weeks [39.7–41.3], PNA 3.9 weeks [0.3–18.1]), 4 pediatric (PNA 4.1 years [1.1–7.4]) and 8 adult liver samples. A relationship with age was found for BCRP, BSEP, GLUT1, MDR1, MRP1, MRP2, MRP3, NTCP, OATP1B1 and OCT1, with the strongest relationship for postmenstrual age. For most transporters mRNA and protein expression were not correlated. No genotype-protein expression relationship was detected. Various developmental patterns of protein expression of hepatic transporters emerged in fetuses and newborns up to four months of age. Postmenstrual age was the most robust factor predicting transporter expression in this cohort. Our data fill an important gap in current pediatric transporter ontogeny knowledge. [Display omitted]