title: The use of stable-isotopically labeled oleic acid to interrogate lipid assembly in vivo: assessing pharmacological effects in preclinical species.
abstract: The use of stable isotopically labeled substrates and analysis by mass spectrometry have provided substantial insight into rates of synthesis, disposition, and utilization of lipids in vivo. The information to be gained from such studies is of particular benefit to therapeutic research where the underlying causes of disease may be related to the production and utilization of lipids. When studying biology through the use of isotope tracers, care must be exercised in interpreting the data to ensure that any response observed can truly be interpreted as biological and not as an artifact of the experimental design or a dilutional effect on the isotope. We studied the effects of dosing route and tracer concentration on the mass isotopomer distribution profile as well as the action of selective inhibitors of microsomal tri-glyceride transfer protein (MTP) in mice and diacylglycerol acyltransferase 1 (DGAT1) in nonhuman primates, using a stable-isotopically labeled approach. Subjects were treated with inhibitor and subsequently given a dose of uniformly ¹³C-labeled oleic acid. Samples were analyzed using a rapid LC-MS technique, allowing the effects of the intervention on the assembly and disposition of triglycerides, cholesteryl esters, and phospholipids to be determined in a single 3 min run from just 10 μl of plasma.
authors.affiliation: Merck Research Laboratories, Merck & Co., Inc. Rahway, NJ. Electronic address: David_McLaren@merck.com.
authors.forename: David G
authors.initials: DG
authors.lastname: McLaren
authors.affiliation: Merck Research Laboratories, Merck & Co., Inc. Rahway, NJ.
authors.forename: Timothy
authors.initials: T
authors.lastname: He
authors.affiliation: Merck Research Laboratories, Merck & Co., Inc. Rahway, NJ.
authors.forename: Sheng-Ping
authors.initials: SP
authors.lastname: Wang
authors.affiliation: Merck Research Laboratories, Merck & Co., Inc. Rahway, NJ.
authors.forename: Vivienne
authors.initials: V
authors.lastname: Mendoza
authors.affiliation: Merck Research Laboratories, Merck & Co., Inc. Rahway, NJ.
authors.forename: Raymond
authors.initials: R
authors.lastname: Rosa
authors.affiliation: Merck Research Laboratories, Merck & Co., Inc. Rahway, NJ.
authors.forename: Karen
authors.initials: K
authors.lastname: Gagen
authors.affiliation: Merck Research Laboratories, Merck & Co., Inc. Rahway, NJ.
authors.forename: Gowri
authors.initials: G
authors.lastname: Bhat
authors.affiliation: Merck Research Laboratories, Merck & Co., Inc. Rahway, NJ.
authors.forename: Kithsiri
authors.initials: K
authors.lastname: Herath
authors.affiliation: Merck Research Laboratories, Merck & Co., Inc. Rahway, NJ.
authors.forename: Paul L
authors.initials: PL
authors.lastname: Miller
authors.affiliation: Merck Research Laboratories, Merck & Co., Inc. Rahway, NJ.
authors.forename: Sloan
authors.initials: S
authors.lastname: Stribling
authors.affiliation: Merck Research Laboratories, Merck & Co., Inc. Rahway, NJ.
authors.forename: Andrew
authors.initials: A
authors.lastname: Taggart
authors.affiliation: Merck Research Laboratories, Merck & Co., Inc. Rahway, NJ.
authors.forename: Jason
authors.initials: J
authors.lastname: Imbriglio
authors.affiliation: Merck Research Laboratories, Merck & Co., Inc. Rahway, NJ.
authors.forename: Jinqi
authors.initials: J
authors.lastname: Liu
authors.affiliation: Merck Research Laboratories, Merck & Co., Inc. Rahway, NJ.
authors.forename: Dunlu
authors.initials: D
authors.lastname: Chen
authors.affiliation: Merck Research Laboratories, Merck & Co., Inc. Rahway, NJ.
authors.forename: Shirly
authors.initials: S
authors.lastname: Pinto
authors.affiliation: Merck Research Laboratories, Merck & Co., Inc. Rahway, NJ.
authors.forename: James M
authors.initials: JM
authors.lastname: Balkovec
authors.affiliation: Merck Research Laboratories, Merck & Co., Inc. Rahway, NJ.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: DeVita
authors.affiliation: Merck Research Laboratories, Merck & Co., Inc. Rahway, NJ.
authors.forename: Donald J
authors.initials: DJ
authors.lastname: Marsh
authors.affiliation: Merck Research Laboratories, Merck & Co., Inc. Rahway, NJ.
authors.forename: Jose M
authors.initials: JM
authors.lastname: Castro-Perez
authors.affiliation: Merck Research Laboratories, Merck & Co., Inc. Rahway, NJ.
authors.forename: Alison
authors.initials: A
authors.lastname: Strack
authors.affiliation: Merck Research Laboratories, Merck & Co., Inc. Rahway, NJ.
authors.forename: Douglas G
authors.initials: DG
authors.lastname: Johns
authors.affiliation: Merck Research Laboratories, Merck & Co., Inc. Rahway, NJ.
authors.forename: Stephen F
authors.initials: SF
authors.lastname: Previs
authors.affiliation: Merck Research Laboratories, Merck & Co., Inc. Rahway, NJ.
authors.forename: Brian K
authors.initials: BK
authors.lastname: Hubbard
authors.affiliation: Merck Research Laboratories, Merck & Co., Inc. Rahway, NJ.
authors.forename: Thomas P
authors.initials: TP
authors.lastname: Roddy
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002522:Chlorocebus aethiops; D002788:Cholesterol Esters / Q000097:blood*; D002853:Chromatography, Liquid; D051048:Diacylglycerol O-Acyltransferase / Q000378:metabolism*; D004333:Drug Administration Routes; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D007553:Isotope Labeling / Q000379:methods; D007554:Isotopes / Q000032:analysis / Q000097:blood; D050356:Lipid Metabolism; D008074:Lipoproteins / Q000097:blood*; D008297:Male; D013058:Mass Spectrometry; D051379:Mice; D008810:Mice, Inbred C57BL; D019301:Oleic Acid / Q000378:metabolism / Q000494:pharmacology; D014280:Triglycerides / Q000097:blood*
chemical_list: D002352:Carrier Proteins; D002788:Cholesterol Esters; D004791:Enzyme Inhibitors; D007554:Isotopes; D008074:Lipoproteins; D014280:Triglycerides; C106092:microsomal triglyceride transfer protein; D019301:Oleic Acid; D051048:Diacylglycerol O-Acyltransferase