title: Multiple microRNA regulation of lipoprotein lipase gene abolished by 3'UTR polymorphisms in a triglyceride-lowering haplotype harboring p.Ser474Ter.
abstract: BACKGROUND
  Lipoprotein lipase (LPL) is a key enzyme in triglyceride (TG) metabolism. LPL gene single nucleotide polymorphisms (SNPs) are associated with TG concentrations however the functionality of many of these SNPs remains poorly understood. MicroRNAs (miR) exert post-transcriptional down-regulation and their target sequence on the 3'UTR may be altered by SNPs. We therefore investigated whether LPL 3'UTR SNPs could modulate plasma TG concentration through the alteration of miR binding-sites.
  
  
  RESULTS
  We performed genetic association studies of LPL 3'UTR SNPs with TG concentrations in 271 type 2 diabetic patients and in general population samples (2997 individuals). A specific LPL haplotype (Hap4) was associated with lower plasma TG concentration (TG-0.18, IC95% [-0.30, -0.07] mmol/L or logTG-0.13, IC95% [-0.18, -0.08], p = 4.77·10(-8)) in the meta-analysis. Hap4 comprises seven 3'UTR SNP minor alleles and p.Ser474Ter (rs328) a well-documented nonsense mutation associated with low TG concentration although by an unknown mechanism so far. Bio-informatic studies identified several putative miRNA binding-sites on the wild-type Hap1 haplotype, lost on Hap4. Functional validation performed in HEK-293T cells using luciferase expression constructs with various LPL 3'UTR allele combinations demonstrated a binding of miR-29, miR-1277 and miR-410 on Hap1, lost on Hap4. This loss of specific miR binding-site in presence of Hap4 was independent of the allelic variation of p.Ser474Ter (rs328).
  
  
  CONCLUSIONS
  We report the regulation of LPL by the miR-29, miR-1277 and miR-410 that is lost in presence of Hap4, a specific LPL TG-lowering haplotype. Consequently p.Ser474Ter association with TG concentration could be at least partially explained by its strong linkage disequilibrium with these functional 3'UTR SNPs.
authors.affiliation: Hospices Civils de Lyon, Hôpital Louis Pradel, Fédération d'endocrinologie, maladies métaboliques, diabète et nutrition, 28 Avenue Doyen Lépine, Bron Cedex F-69677, France; INSERM U1060, Laboratoire Carmen, Université Lyon 1, INRA U1235, INSA de Lyon, Bâtiment IMBL, INSA-Lyon, 11 Avenue Jean Capelle, 69621 Villeurbanne Cedex, France.
authors.forename: Cyrielle
authors.initials: C
authors.lastname: Caussy
authors.affiliation: Hospices Civils de Lyon, Hôpital Louis Pradel, Fédération d'endocrinologie, maladies métaboliques, diabète et nutrition, 28 Avenue Doyen Lépine, Bron Cedex F-69677, France; INSERM U1060, Laboratoire Carmen, Université Lyon 1, INRA U1235, INSA de Lyon, Bâtiment IMBL, INSA-Lyon, 11 Avenue Jean Capelle, 69621 Villeurbanne Cedex, France. Electronic address: sybil.charriere@chu-lyon.fr.
authors.forename: Sybil
authors.initials: S
authors.lastname: Charrière
authors.affiliation: INSERM, U1167, Institut Pasteur de Lille, Université de Lille, 1 rue du Pr. Calmette, BP 245, F-59019 Lille Cedex, France.
authors.forename: Aline
authors.initials: A
authors.lastname: Meirhaeghe
authors.affiliation: INSERM, U1167, Institut Pasteur de Lille, Université de Lille, 1 rue du Pr. Calmette, BP 245, F-59019 Lille Cedex, France.
authors.forename: Jean
authors.initials: J
authors.lastname: Dallongeville
authors.affiliation: INSERM U1060, Laboratoire Carmen, Université Lyon 1, INRA U1235, INSA de Lyon, Bâtiment IMBL, INSA-Lyon, 11 Avenue Jean Capelle, 69621 Villeurbanne Cedex, France.
authors.forename: Etienne
authors.initials: E
authors.lastname: Lefai
authors.affiliation: INSERM U1060, Laboratoire Carmen, Université Lyon 1, INRA U1235, INSA de Lyon, Bâtiment IMBL, INSA-Lyon, 11 Avenue Jean Capelle, 69621 Villeurbanne Cedex, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Rome
authors.affiliation: INSERM U1060, Laboratoire Carmen, Université Lyon 1, INRA U1235, INSA de Lyon, Bâtiment IMBL, INSA-Lyon, 11 Avenue Jean Capelle, 69621 Villeurbanne Cedex, France; Hospices Civils de Lyon, Centre Hospitalier Lyon-Sud, Centre de Biologie Sud, Laboratoire de Biochimie moléculaire et métabolique, 165 chemin du Grand Revoyet, Pierre-Bénite Cedex F-69495, France.
authors.forename: Charlotte
authors.initials: C
authors.lastname: Cuerq
authors.affiliation: INSERM U1060, Laboratoire Carmen, Université Lyon 1, INRA U1235, INSA de Lyon, Bâtiment IMBL, INSA-Lyon, 11 Avenue Jean Capelle, 69621 Villeurbanne Cedex, France.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Euthine
authors.affiliation: Hospices Civils de Lyon, Centre Hospitalier Lyon-Sud, Centre de Biologie Sud, Laboratoire de Biochimie moléculaire et métabolique, 165 chemin du Grand Revoyet, Pierre-Bénite Cedex F-69495, France.
authors.forename: Mireille
authors.initials: M
authors.lastname: Delay
authors.affiliation: INSERM U1060, Laboratoire Carmen, Université Lyon 1, INRA U1235, INSA de Lyon, Bâtiment IMBL, INSA-Lyon, 11 Avenue Jean Capelle, 69621 Villeurbanne Cedex, France; Hospices Civils de Lyon, Centre de Biologie et de Pathologie Est, Département de biochimie et biologie moléculaire, 59 Boulevard Pinel, Bron Cedex F-69677, France.
authors.forename: Oriane
authors.initials: O
authors.lastname: Marmontel
authors.affiliation: INSERM U1060, Laboratoire Carmen, Université Lyon 1, INRA U1235, INSA de Lyon, Bâtiment IMBL, INSA-Lyon, 11 Avenue Jean Capelle, 69621 Villeurbanne Cedex, France; Hospices Civils de Lyon, Centre de Biologie et de Pathologie Est, Département de biochimie et biologie moléculaire, 59 Boulevard Pinel, Bron Cedex F-69677, France.
authors.forename: Mathilde
authors.initials: M
authors.lastname: Di Filippo
authors.affiliation: INSERM U1060, Laboratoire Carmen, Université Lyon 1, INRA U1235, INSA de Lyon, Bâtiment IMBL, INSA-Lyon, 11 Avenue Jean Capelle, 69621 Villeurbanne Cedex, France.
authors.forename: Michel
authors.initials: M
authors.lastname: Lagarde
authors.affiliation: Hospices Civils de Lyon, Hôpital Louis Pradel, Fédération d'endocrinologie, maladies métaboliques, diabète et nutrition, 28 Avenue Doyen Lépine, Bron Cedex F-69677, France; INSERM U1060, Laboratoire Carmen, Université Lyon 1, INRA U1235, INSA de Lyon, Bâtiment IMBL, INSA-Lyon, 11 Avenue Jean Capelle, 69621 Villeurbanne Cedex, France; CENS, Centre de Recherche en Nutrition Humaine Rhône-Alpes, Centre Hospitalier Lyon Sud, 165 chemin du Grand Revoyet, 69310 Pierre-Bénite, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Moulin
authors.affiliation: INSERM U1060, Laboratoire Carmen, Université Lyon 1, INRA U1235, INSA de Lyon, Bâtiment IMBL, INSA-Lyon, 11 Avenue Jean Capelle, 69621 Villeurbanne Cedex, France; Hospices Civils de Lyon, Centre Hospitalier Lyon-Sud, Centre de Biologie Sud, Laboratoire de Biochimie moléculaire et métabolique, 165 chemin du Grand Revoyet, Pierre-Bénite Cedex F-69495, France; CENS, Centre de Recherche en Nutrition Humaine Rhône-Alpes, Centre Hospitalier Lyon Sud, 165 chemin du Grand Revoyet, 69310 Pierre-Bénite, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Marçais
mesh_terms: D020413:3' Untranslated Regions; D001665:Binding Sites; D015415:Biomarkers / Q000097:blood; D016022:Case-Control Studies; D019295:Computational Biology; D030541:Databases, Genetic; D003924:Diabetes Mellitus, Type 2 / Q000097:blood / Q000175:diagnosis; D015971:Gene Expression Regulation, Enzymologic; D005787:Gene Frequency; D056726:Genetic Association Studies; D020022:Genetic Predisposition to Disease; D057809:HEK293 Cells; D006239:Haplotypes; D006801:Humans; D015228:Hypertriglyceridemia / Q000097:blood / Q000175:diagnosis / Q000201:enzymology / Q000235:genetics*; D015810:Linkage Disequilibrium; D008071:Lipoprotein Lipase / Q000235:genetics* / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D010641:Phenotype; D020641:Polymorphism, Single Nucleotide; D012307:Risk Factors; D014162:Transfection; D014280:Triglycerides / Q000097:blood*
chemical_list: D020413:3' Untranslated Regions; D015415:Biomarkers; C532045:MIRN29a microRNA, human; C568527:MIRN410 microRNA, human; D035683:MicroRNAs; D014280:Triglycerides; C548261:LPL protein, human; D008071:Lipoprotein Lipase
keywords: Haplotype; Lipids; Lipoprotein lipase; MicroRNA; SNP; Triglycerides