PCSK9 Induces CD36 Degradation and Affects Long-Chain Fatty Acid Uptake and Triglyceride Metabolism in Adipocytes and in Mouse Liver.
OBJECTIVE
Proprotein convertase subtilisin/kexin type 9 (PCSK9) promotes the degradation of the low-density lipoprotein receptor thereby elevating plasma low-density lipoprotein cholesterol levels and the risk of coronary heart disease. Thus, the use of PCSK9 inhibitors holds great promise to prevent heart disease. Previous work found that PCSK9 is involved in triglyceride metabolism, independently of its action on low-density lipoprotein receptor, and that other yet unidentified receptors could mediate this effect. Therefore, we assessed whether PCSK9 enhances the degradation of CD36, a major receptor involved in transport of long-chain fatty acids and triglyceride storage.


RESULTS
Overexpressed or recombinant PCSK9 induced CD36 degradation in cell lines and primary adipocytes and reduced the uptake of the palmitate analog Bodipy FL C16 and oxidized low-density lipoprotein in 3T3-L1 adipocytes and hepatic HepG2 cells, respectively. Surface plasmon resonance, coimmunoprecipitation, confocal immunofluorescence microscopy, and protein degradation pathway inhibitors revealed that PCSK9 directly interacts with CD36 and targets the receptor to lysosomes through a mechanism involving the proteasome. Importantly, the level of CD36 protein was increased by >3-fold upon small interfering RNA knockdown of endogenous PCSK9 in hepatic cells and similarly increased in the liver and visceral adipose tissue of Pcsk9(-/-) mice. In Pcsk9(-/-) mice, increased hepatic CD36 was correlated with an amplified uptake of fatty acid and accumulation of triglycerides and lipid droplets.


CONCLUSIONS
Our results demonstrate an important role of PCSK9 in modulating the function of CD36 and triglyceride metabolism. PCSK9-mediated CD36 degradation may serve to limit fatty acid uptake and triglyceride accumulation in tissues, such as the liver.
From the Laboratory of Molecular Cell Biology (A.D., S.S., E.T.N.S., G.M.) and Laboratory of Metabolomic (C.D.R.), Montreal Heart Institute, Montréal, Québec, Canada; Université de Nantes, L'institut du thorax, Inserm UMR 1087 / CNRS UMR 6291, Nantes, France (B.L.); and Faculty of Pharmacy (H.O.), Université de Montréal, Department of Pharmacology, Faculty of Medicine (S.S., E.T.N.S., G.M.), Department of Nutrition, Faculty of Medicine (C.D.R.), and Department of Medicine (G.M.), Faculty of Medicine, Université de Montréal, Montréal, Québec, Canada.
Annie
A
Demers
From the Laboratory of Molecular Cell Biology (A.D., S.S., E.T.N.S., G.M.) and Laboratory of Metabolomic (C.D.R.), Montreal Heart Institute, Montréal, Québec, Canada; Université de Nantes, L'institut du thorax, Inserm UMR 1087 / CNRS UMR 6291, Nantes, France (B.L.); and Faculty of Pharmacy (H.O.), Université de Montréal, Department of Pharmacology, Faculty of Medicine (S.S., E.T.N.S., G.M.), Department of Nutrition, Faculty of Medicine (C.D.R.), and Department of Medicine (G.M.), Faculty of Medicine, Université de Montréal, Montréal, Québec, Canada.
Samaneh
S
Samami
From the Laboratory of Molecular Cell Biology (A.D., S.S., E.T.N.S., G.M.) and Laboratory of Metabolomic (C.D.R.), Montreal Heart Institute, Montréal, Québec, Canada; Université de Nantes, L'institut du thorax, Inserm UMR 1087 / CNRS UMR 6291, Nantes, France (B.L.); and Faculty of Pharmacy (H.O.), Université de Montréal, Department of Pharmacology, Faculty of Medicine (S.S., E.T.N.S., G.M.), Department of Nutrition, Faculty of Medicine (C.D.R.), and Department of Medicine (G.M.), Faculty of Medicine, Université de Montréal, Montréal, Québec, Canada.
Benjamin
B
Lauzier
From the Laboratory of Molecular Cell Biology (A.D., S.S., E.T.N.S., G.M.) and Laboratory of Metabolomic (C.D.R.), Montreal Heart Institute, Montréal, Québec, Canada; Université de Nantes, L'institut du thorax, Inserm UMR 1087 / CNRS UMR 6291, Nantes, France (B.L.); and Faculty of Pharmacy (H.O.), Université de Montréal, Department of Pharmacology, Faculty of Medicine (S.S., E.T.N.S., G.M.), Department of Nutrition, Faculty of Medicine (C.D.R.), and Department of Medicine (G.M.), Faculty of Medicine, Université de Montréal, Montréal, Québec, Canada.
Christine
C
Des Rosiers
From the Laboratory of Molecular Cell Biology (A.D., S.S., E.T.N.S., G.M.) and Laboratory of Metabolomic (C.D.R.), Montreal Heart Institute, Montréal, Québec, Canada; Université de Nantes, L'institut du thorax, Inserm UMR 1087 / CNRS UMR 6291, Nantes, France (B.L.); and Faculty of Pharmacy (H.O.), Université de Montréal, Department of Pharmacology, Faculty of Medicine (S.S., E.T.N.S., G.M.), Department of Nutrition, Faculty of Medicine (C.D.R.), and Department of Medicine (G.M.), Faculty of Medicine, Université de Montréal, Montréal, Québec, Canada.
Emilienne Tudor
ET
Ngo Sock
From the Laboratory of Molecular Cell Biology (A.D., S.S., E.T.N.S., G.M.) and Laboratory of Metabolomic (C.D.R.), Montreal Heart Institute, Montréal, Québec, Canada; Université de Nantes, L'institut du thorax, Inserm UMR 1087 / CNRS UMR 6291, Nantes, France (B.L.); and Faculty of Pharmacy (H.O.), Université de Montréal, Department of Pharmacology, Faculty of Medicine (S.S., E.T.N.S., G.M.), Department of Nutrition, Faculty of Medicine (C.D.R.), and Department of Medicine (G.M.), Faculty of Medicine, Université de Montréal, Montréal, Québec, Canada.
Huy
H
Ong
From the Laboratory of Molecular Cell Biology (A.D., S.S., E.T.N.S., G.M.) and Laboratory of Metabolomic (C.D.R.), Montreal Heart Institute, Montréal, Québec, Canada; Université de Nantes, L'institut du thorax, Inserm UMR 1087 / CNRS UMR 6291, Nantes, France (B.L.); and Faculty of Pharmacy (H.O.), Université de Montréal, Department of Pharmacology, Faculty of Medicine (S.S., E.T.N.S., G.M.), Department of Nutrition, Faculty of Medicine (C.D.R.), and Department of Medicine (G.M.), Faculty of Medicine, Université de Montréal, Montréal, Québec, Canada. gaetan.mayer@icm-mhi.org.
Gaetan
G
Mayer
D041721:3T3-L1 Cells; D017667:Adipocytes / Q000187:drug effects / Q000201:enzymology*; D000818:Animals; D001896:Boron Compounds / Q000378:metabolism; D018955:CD36 Antigens / Q000235:genetics / Q000378:metabolism*; D005227:Fatty Acids / Q000378:metabolism*; D005260:Female; D057809:HEK293 Cells; D056945:Hep G2 Cells; D006801:Humans; D050152:Intra-Abdominal Fat / Q000187:drug effects / Q000201:enzymology*; D008077:Lipoproteins, LDL / Q000378:metabolism; D008099:Liver / Q000187:drug effects / Q000201:enzymology*; D008247:Lysosomes / Q000201:enzymology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010169:Palmitic Acids / Q000378:metabolism; D000071449:Proprotein Convertase 9; D043484:Proprotein Convertases / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D061988:Proteasome Inhibitors / Q000494:pharmacology; D059748:Proteolysis; D034622:RNA Interference; D012697:Serine Endopeptidases / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors; D014162:Transfection; D014280:Triglycerides / Q000378:metabolism*
C543305:4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-hexadecanoic acid; D001896:Boron Compounds; D018955:CD36 Antigens; D005227:Fatty Acids; D008077:Lipoproteins, LDL; D010169:Palmitic Acids; D061988:Proteasome Inhibitors; D014280:Triglycerides; C099252:oxidized low density lipoprotein; C472125:PCSK9 protein, human; C472126:Pcsk9 protein, mouse; D000071449:Proprotein Convertase 9; D043484:Proprotein Convertases; D012697:Serine Endopeptidases; D046988:Proteasome Endopeptidase Complex
Pcsk9 protein, mouse; antigens, CD36; mice, knockout; receptors, LDL; triglyceride