PROS AND CONS OF CHOLINE SUPPLEMENTATION. Steven H. Zeisel, MD, PhD Professor of Nutrition and Pediatrics University of North Carolina at Chapel Hill Director, UNC Nutrition Research Institute Director, UNC Nutrition Obesity Research Center steven_zeisel@unc.edu
STEVEN ZEISEL DISCLOSURES AFFILIATION/FINANCIAL INTERESTS Grants/Research Support: Scientific Advisory Board/Consultant: Honorarium for this talk: CORPORATE ORGANIZATION NIH, USDA, Nestle, Hershey, Egg Nutrition Center Dupont Nutrition, Metabolon, Nutrigene Sciences, Genovive, American Pistachio Growers. Wyeth (Nestle) Stock Shareholder: Nutrigene Sciences
CHOLINE IOM designated as essential nutrient in 1998 Needed for membranes, acetylcholine, methylation Important for liver and muscle function Critical for fetal brain development
mg/100g food 400 300 200 100 FOOD CHOLINE Daily requirement estimated at 550 mg From http://www.ars.usda.gov/services/docs.htm?docid=6232
MOST AMERICANS DO NOT EAT ENOUGH CHOLINE Males AI Females pregnancy AI mg/day
ENDOGENOUS SYNTHESIS OF CHOLINE Phosphatidylethanolamine N- methyltransferase (PEMT) 3 S-adenosylmethionine + phosphatidylethanolamine phosphatidylcholine FASEB J 21:2622-32, 2007
DIETARY REQUIREMENT FOR CHOLINE 77% of Men 80% of Postmenopausal Women 44% of Premenopausal Women Need to Eat Choline (fatty liver, muscle damage)
IN LOW CHOLINE FATTY LIVER DEVELOPS Mean Liver/Spleen CPK Level fat (U/L) ratio 5000 3.0 4500 4000 2.25 3500 3000 2500 1.5 2000 1500 0.75 1000 500 0 * Normal Diet Depletion Repletion Phase Of Study Am J Clin Nutr 2007;85:1275-1285
EXPORT OF FAT FROM LIVER REQUIRES CHOLINE triacylglycerol phosphatidylcholine apoprotein VLDL
ARE SOME PEOPLE ESPECIALLY SENSITIVE TO DIETARY CHOLINE? Single nucleotide polymorphisms (SNPs)
SNPS CAUSE METABOLIC INEFFICIENCIES Metabolism Nutrient Metabolite No SNP Nutrient Metabolite metabolite Metabolite SNP DIET DELIVERS METABOLITE
NAFLD PREDICTED BY CHOLINE SNPs Fatty liver Lowest Highest Group Group 0 1 2 Variant alleles triacylglycerol phosphatidylcholine apoprotein VLDL FASEB J. 2013 27:1674-89
LOW CHOLINE IN 10% OF PEOPLE RESULTS IN MUSCLE DAMAGE Mean CPK Level (U/L) 2500 5000 4500 4000 1875 3500 3000 1250 2500 2000 1500 625 1000 500 0 * Normal Diet Depletion Repletion Phase Of Study American Journal of Clinical Nutrition 85(5):1275-85, 2007
LOW CHOLINE IN 10% OF PEOPLE RESULTS IN MUSCLE DAMAGE FASEB J 2014;28:2970-2978 FASEB J. 2014 28:2970-8.
Europe Asia Mexico Africa AA AG GG Need more choline rs2771040 SLC44A1 A G
Europe Asia Mexico Africa GG GA AA rs1557502 Need more choline CHKB G A
REQUIREMENT FOR CHOLINE - GENDER 77% of Men 80% of Postmenopausal Women 44% of Premenopausal Women WHY DON T 56% OF YOUNG WOMEN NEED CHOLINE? V
ENDOGENOUS SYNTHESIS OF CHOLINE INDUCED BY ESTROGEN Phosphatidylethanolamine N- methyltransferase (PEMT) + 3 S-adenosylmethionine + phosphatidylethanolamine phosphatidylcholine Estrogen
PEMT INDUCED BY ESTROGEN Mouse Rat hepatocyte Human RNA RNA Activity Activity FASEB Journal 21:2622-32, 2007
REQUIREMENT FOR CHOLINE - GENDER IF THEY COULD MAKE THEIR OWN CHOLINE, WHY DO 44% OF YOUNG WOMEN STILL NEED CHOLINE? V
SNP PEMT rs12325817 PREVENTS INDUCTION BY ESTROGEN Phosphatidylethanolamine N- methyltransferase (PEMT) + 3 S-adenosylmethionine + phosphatidylethanolamine X phosphatidylcholine Estrogen J. Biol. Chem. 286:1649-58, 2011
PEMT SNP RESULTS IN NO RESPONSE TO ESTROGEN No alleles (GG) 1 allele (GC) 2 alleles (CC) Phosphatidylethanolamine N- methyltransferase (PEMT) X 3 S-adenosylmethionine + phosphatidylcholine phosphatidylethanolamine Primary human hepatocytes. Rs12325817; Rx moxesterol for 24 h. J. Biol. Chem. 286:1649-58, 2011
PEMT rs12325817 FREQUENCY Europe Asia Mexico Africa GG GC CC rs12325817 Need more choline PEMT G C
CHOLINE IN PREGNANCY & LACTATION Maternal blood = 10 µm Amniotic fluid = 140 µm Mediated uptake of choline across the placenta. Breast milk rich in choline compounds 1500 µm Mediated transport of choline across the mammary gland. Graphics: www.npr.org; www.wholeearthherbals.com
SERUM CHOLINE HIGH DURING FIRST YEAR IN HUMANS 35 Serum Free Choline, µmol/l 30 25 20 15 10 5 Adult level 0 Postnatal day Ilcol et al., J Nutr Biochem. 2005;16:489-99.
26 24 CHOLINE DURING Choices BRAIN Before DEVELOPMENT 20 IMPROVES Maze is MEMORY Empty 18 CRITICAL PERIODS 22 OF 16 14 1 2 3 4 Blocks of 3 Sessions control prenatal 6-11 prenatal 12-17 postnatal 1-15 postnatal 16-30 postnatal 31-45 dams 200 mg/day in water pups 300 mg/kg sc From Meck and Williams Neurosci Biobehav Rev. 27(4):385-99, 2003
LIFELONG EFFECT OF CHOLINE Neurosci Biobehav Rev. 27:385-99, 2003
1 st & 2 nd TRIMESTER MATERNAL CHOLINE INTAKE PREDICTS MEMORY IN CHILD AT 7 Yr Wide Range Assessment of Memory and Learning, Second Edition (WRAML2), Design and Picture Memory subtests at age 7 y among 890 mother-child pairs in Project Viva, eastern Massachusetts, 1999 2002 to 2008 2011. Replotted from: Boeke C E et al. Am. J. Epidemiol. 2013;177:1338-1347
PRE- & POSTNATAL CRITICAL PERIOD FOR CHOLINE Rodent gives birth on day E21 - Hippocampus develops E12-E18 Human equivalent = ~25 weeks gestation - 4 yrs postnatal
MATERNAL CHOLINE MODULATES HOW FETAL NEURAL PROGENITORS DIVIDE Low Control
NEUROGENESIS IS MODULATED BY CHOLINE E17 Neural progenitors in fetal brain were reduced if mother (mouse) fed a low vs. control choline diet during pregnancy. Low choline Normal choline Nestin-CFPnuc transgenic mice
NEUROGENESIS IS MODULATED BY CHOLINE CORTEX E17 Late-born neurons missing in cortex layers 2-6
NEUROGENESIS IS ALTERED BY EPIDERMAL GROWTH FACTOR RECEPTOR (EGFR) Activation of the EGF receptor in late progenitor cells stimulates cell proliferation, migration, and astrocyte differentiation. Differences in the level of EGFR expression determine how progenitor cells interpret an extrinsic signal at specific stages of development. EGFR is upregulated at midgestation in the mouse telencephalon. From www.astrochem.org
LOW CHOLINE ATTENUATES RESPONSE TO EGF pegfr perk1/2 pakt NPCs
EFFECT OF CHOLINE MEDIATED BY EGFR PROTEIN REDUCTION
NPC GROWTH RESTORED BY TRANSFECTION WITH EGFR
LOW CHOLINE DECREASES EGFR SYNTHESIS DESPITE NORMAL mrna Egfr mrna cyclohexamide
IN NPCs LOW CHOLINE ALTERS mir-129 EXPRESSION 3 Fold change 2 1 0 Overexpressed Underexpressed
IN NPCs EGFR DECREASED BY mir-129 EGFR
EFFECT OF LOW CHOLINE MEDIATED BY LOSS OF EGFR
OTHER COMMON SNPS ALTER CHOLINE REQUIREMENT BHMT FASEB J. 20, 1336 1344 (2006); 28:2970-2978 (2014) PROC.NAT.ACAD.SCI. USA 102: 16025-16030 (2005)
CHOLINE AND SPERM DYSFUNCTION FASEB J 2014;28:2970-2978
MEN WITH CHDH rs12676 G233T
CHDH rs12676 G233T DYSMORPHIC MITOCHONDRIA PLoS One. 2012;7(4):e36047. doi: 10.1371/journal.pone.0036047
CHDH rs12676 G233T LOW ATP CHDH PLoS One. 2012;7(4):e36047. doi: 10.1371/journal.pone.0036047
Europe Asia Mexico Africa GG GT TT rs12676 CHDH G T Need more choline
LOW CHOLINE (METHYL DEFICIENT) DIET RESULTS IN LIVER CANCER 80 70 Number tumors 60 50 40 30 20 53% 100% Cancer Nodules 10 0 Low 0% Ctl N=45 N=45 Ghoshal &Farber. Carcinogenesis 5:1367-70, 1984
DELETION OF Bhmt RESULTS IN LIVER CANCER BHM T
AT 1 YR DELETION OF Bhmt RESULTS IN LIVER CANCER J Biol Chem. 2011;286:36258-67
DO HUMANS WITH BHMT SNPS GET LIVER CANCER??? Europe Asia Mexico Africa GG GA AA Need more choline rs3733890 BHMT G A
SUMMARY: CHOLINE & HEALTH Subpopulations (SNPs) with special needs. Fatty Liver Muscle Critical nutrient for brain development Sperm function Cancer
WILL CHOLINE STORY LAST? Year 2364, Star Trek Next Generation
Thank you A. Johnson K-A. da Costa M. Spencer Y. Wang Y-W. Teng L. Fischer E. Yen J. Owen M. Niculescu K. Corbin C. Craciunescu N. Surzenko M. Mehedint M. Resseguie H-M. Hwang S. Orena W. Sha M. Kohlmeier W. Friday E. Pjetri X. Zhu J. Song X. Zhou D. Lupu S. Goodson Collaborators: Jan Blusztajn, Tina Williams, Wei Jia, Tim Garrow, Joe Galanko Funding: NIH (NIDDK, ODS, NIA), USDA, Bill & Melinda Gates Foundation, Nestle.