title: An APOA5 3' UTR variant associated with plasma triglycerides triggers APOA5 downregulation by creating a functional miR-485-5p binding site.
abstract: APOA5 c.*158C>T (rs2266788), located in the 3' UTR, belongs to APOA5 haplotype 2 (APOA5*2), which is strongly associated with plasma triglyceride levels and modulates the occurrence of both moderate and severe hypertriglyceridemia. Individuals with APOA5*2 display reduced APOA5 expression at the posttranscriptional level. However, the functionality of this haplotype remains unclear. We hypothesized that the hypertriglyceridemic effects of APOA5*2 could involve miRNA regulation in the APOA5 3' UTR. Bioinformatic studies have identified the creation of a potential miRNA binding site for liver-expressed miR-485-5p (MIRN485-5p) in the mutant APOA5 3' UTR with the c.*158C allele. In human embryonic kidney 293T (HEK293T) cells cotransfected with an APOA5 3' UTR luciferase reporter vector and a miR485-5p precursor, c.*158C allele expression was significantly decreased. Moreover, in HuH-7 cells endogenously expressing miR-485-5p, we observed that luciferase activity was significantly lower in the presence of the c.*158C allele than in the presence of the c.*158T allele, which was completely reversed by a miR-485-5p inhibitor. We demonstrated that the rare c.*158C APOA5 allele creates a functional target site for liver-expressed miR-485-5p. Therefore, we propose that the well-documented hypertriglyceridemic effect of APOA5*2 involves an APOA5 posttranscriptional downregulation mediated by miR-485-5p.
authors.affiliation: Fédération d'Endocrinologie, Maladies Métaboliques, Diabète, et Nutrition, Hôpital Louis Pradel, Hospices Civils de Lyon, 69677 Bron Cedex, France; Laboratoire Carmen, Institut National de la Santé et de la Recherche Médicale U1060, Institut National de la Recherche Agronomique U1235, Institut National des Sciences Appliquées de Lyon, Université Claude Bernard Lyon 1, 69621 Villeurbanne Cedex, France.
authors.forename: Cyrielle
authors.initials: C
authors.lastname: Caussy
authors.affiliation: Fédération d'Endocrinologie, Maladies Métaboliques, Diabète, et Nutrition, Hôpital Louis Pradel, Hospices Civils de Lyon, 69677 Bron Cedex, France; Laboratoire Carmen, Institut National de la Santé et de la Recherche Médicale U1060, Institut National de la Recherche Agronomique U1235, Institut National des Sciences Appliquées de Lyon, Université Claude Bernard Lyon 1, 69621 Villeurbanne Cedex, France. Electronic address: sybil.charriere@chu-lyon.fr.
authors.forename: Sybil
authors.initials: S
authors.lastname: Charrière
authors.affiliation: Laboratoire Carmen, Institut National de la Santé et de la Recherche Médicale U1060, Institut National de la Recherche Agronomique U1235, Institut National des Sciences Appliquées de Lyon, Université Claude Bernard Lyon 1, 69621 Villeurbanne Cedex, France; Laboratoire de Biochimie Moléculaire et Métabolique, Centre de Biologie Sud, Centre Hospitalier Lyon-Sud, Hospices Civils de Lyon, 69495 Pierre-Bénite Cedex, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Marçais
authors.affiliation: Laboratoire Carmen, Institut National de la Santé et de la Recherche Médicale U1060, Institut National de la Recherche Agronomique U1235, Institut National des Sciences Appliquées de Lyon, Université Claude Bernard Lyon 1, 69621 Villeurbanne Cedex, France; Département de Biochimie et Biologie Moléculaire, Centre de Biologie et de Pathologie Est, Hospices Civils de Lyon, 69677 Bron Cedex, France.
authors.forename: Mathilde
authors.initials: M
authors.lastname: Di Filippo
authors.affiliation: Laboratoire Carmen, Institut National de la Santé et de la Recherche Médicale U1060, Institut National de la Recherche Agronomique U1235, Institut National des Sciences Appliquées de Lyon, Université Claude Bernard Lyon 1, 69621 Villeurbanne Cedex, France; Département de Biochimie et Biologie Moléculaire, Centre de Biologie et de Pathologie Est, Hospices Civils de Lyon, 69677 Bron Cedex, France.
authors.forename: Agnès
authors.initials: A
authors.lastname: Sassolas
authors.affiliation: Laboratoire de Biochimie Moléculaire et Métabolique, Centre de Biologie Sud, Centre Hospitalier Lyon-Sud, Hospices Civils de Lyon, 69495 Pierre-Bénite Cedex, France.
authors.forename: Mireille
authors.initials: M
authors.lastname: Delay
authors.affiliation: Laboratoire Carmen, Institut National de la Santé et de la Recherche Médicale U1060, Institut National de la Recherche Agronomique U1235, Institut National des Sciences Appliquées de Lyon, Université Claude Bernard Lyon 1, 69621 Villeurbanne Cedex, France.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Euthine
authors.affiliation: Laboratoire Carmen, Institut National de la Santé et de la Recherche Médicale U1060, Institut National de la Recherche Agronomique U1235, Institut National des Sciences Appliquées de Lyon, Université Claude Bernard Lyon 1, 69621 Villeurbanne Cedex, France.
authors.forename: Audrey
authors.initials: A
authors.lastname: Jalabert
authors.affiliation: Laboratoire Carmen, Institut National de la Santé et de la Recherche Médicale U1060, Institut National de la Recherche Agronomique U1235, Institut National des Sciences Appliquées de Lyon, Université Claude Bernard Lyon 1, 69621 Villeurbanne Cedex, France.
authors.forename: Etienne
authors.initials: E
authors.lastname: Lefai
authors.affiliation: Laboratoire Carmen, Institut National de la Santé et de la Recherche Médicale U1060, Institut National de la Recherche Agronomique U1235, Institut National des Sciences Appliquées de Lyon, Université Claude Bernard Lyon 1, 69621 Villeurbanne Cedex, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Rome
authors.affiliation: Fédération d'Endocrinologie, Maladies Métaboliques, Diabète, et Nutrition, Hôpital Louis Pradel, Hospices Civils de Lyon, 69677 Bron Cedex, France; Laboratoire Carmen, Institut National de la Santé et de la Recherche Médicale U1060, Institut National de la Recherche Agronomique U1235, Institut National des Sciences Appliquées de Lyon, Université Claude Bernard Lyon 1, 69621 Villeurbanne Cedex, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Moulin
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics*; D000483:Alleles; D000072040:Apolipoprotein A-V; D001054:Apolipoproteins A / Q000235:genetics* / Q000378:metabolism; D001665:Binding Sites; D019295:Computational Biology; D015536:Down-Regulation; D014644:Genetic Variation; D057809:HEK293 Cells; D006239:Haplotypes; D006801:Humans; D008099:Liver / Q000378:metabolism; D008156:Luciferases / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D014280:Triglycerides / Q000097:blood*
chemical_list: D020413:3' Untranslated Regions; C437704:APOA5 protein, human; D000072040:Apolipoprotein A-V; D001054:Apolipoproteins A; C556501:MIRN485 microRNA, human; D035683:MicroRNAs; D014280:Triglycerides; D008156:Luciferases