Specificity and rate of human and mouse liver and plasma phosphatidylcholine synthesis analyzed in vivo[S]

• Published in: Journal of lipid research Vol. 52; no. 2; pp. 399 - 407
• Main Authors: Pynn, Christopher J., Henderson, Neil G., Clark, Howard, Koster, Grielof, Bernhard, Wolfgang, Postle, Anthony D.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.02.2011; American Society for Biochemistry and Molecular Biology; The American Society for Biochemistry and Molecular Biology; Elsevier
• Subjects: Adult; Animals; Choline - metabolism; Cytidine Diphosphate Choline - metabolism; deuterated choline; Deuterium; Female; Humans; lipidomics; Liver - metabolism; Lung - metabolism; Male; Methods; Mice; multiple isotopomer distribution analysis; Phosphatidylcholines - biosynthesis; Phosphatidylethanolamine N-Methyltransferase - blood; Phosphatidylethanolamine N-Methyltransferase - metabolism; phosphatidylethanolamine-N-methyltransferase; stable isotope; multiple isotopomer distribution analysis; stable isotope; phosphatidylethanolamine-N-methyltransferase; deuterated choline; lipidomics
• ISSN: 0022-2275; 1539-7262; 1539-7262
• DOI: 10.1194/jlr.D011916

Phosphatidylcholine (PC) synthesis by the direct cytidine diphosphate choline (CDP-choline) pathway in rat liver generates predominantly mono- and di-unsaturated molecular species, while polyunsaturated PC species are synthesized largely by the phosphatidylethanolamine-N-methyltransferase (PEMT) pathway. Although altered PC synthesis has been suggested to contribute to development of hepatocarcinoma and nonalcoholic steatohepatitis, analysis of the specificity of hepatic PC metabolism in human patients has been limited by the lack of sensitive and safe methodologies. Here we incorporated a deuterated methyl-d9-labled choline chloride, to quantify biosynthesis fluxes through both of the PC synthetic pathways in vivo in human volunteers and compared these fluxes with those in mice. Rates and molecular specificities of label incorporated into mouse liver and plasma PC were very similar and strongly suggest that label incorporation into human plasma PC can provide a direct measure of hepatic PC synthesis in human subjects. Importantly, we demonstrate for the first time that the PEMT pathway in human liver is selective for polyunsaturated PC species, especially those containing docosahexaenoic acid. Finally, we present a multiple isotopomer distribution analysis approach, based on transfer of deuterated methyl groups to S-adenosylmethionine and subsequent sequential methylations of PE, to quantify absolute flux rates through the PEMT pathway that are applicable to studies of liver dysfunction in clinical studies.