UQÀM: Department of Biological Sciences The role of apolipoprotein D in lipid metabolism and metabolic syndrome Catherine Mounier PhD
Apolipoprotein D Apolipoprotéine D (apod) Secreted protein Associated with HDL Secretion stimulated by 25- hydroxycholesterol Ligands Arachidonic Acid Heme Steroids (progesterone, androgen, estrogen) Cholesterol and CHOL esters
Apolipoprotein D Surrenals Kidney Pancreas Thymus Central and peripheric nervous systems Lungs Ovaries and testis Central and peripheric nervous systems Adipose tissue Heart Lungs Glands Ovaries and testis
Apolipoprotein D Involved in growth and cellular differentiation Metabolic disorders and energetic homeastasis Development Cancer Protection against stress in drosophila Maintance and repair of central nervous system
Apolipoprotein D in metabolism Low plasma levels of apod associated with disorders that produce low HDL levels with or without hypertriglyceridemia Tangier disease Abeta-lipoproteine-mia LCAT deficiency Hypertrigly-ceride-mia at birth ApoD modulation in non-insulin dependent diabetes mellitus (NIDDM) Increased and defective processing of apod in Niemann-Pick disease type C mouse (NPC). Animal model of human NPC, which is a genetic disorder affecting cellular cholesterol transport
Apolipoprotein D Tissue Do Carmo et al, AJP Endo Met 2008
Tg ApoD mice characteristics Table 2. Morphometric parameters of H-apoD Tg mice and WT littermates. WT Thy-1/ApoD NSE/ApoD Body weight (g) 38.95 ± 4.57 44.06 ± 5.42 43.83 ± 4.16 Body lenght (cm) 9.86 ± 0.40 10.11 ± 0.36 10.00 ± 0.18 Body mass index (g/cm2) 0.40 ± 0.03 0.43 ± 0.05 0.44 ± 0.04 Weights of tissue (g/g of body weight, x 100) Brain 1.07 ± 0.08 0.97 ± 0.08 0.96 ± 0.07 Liver 4.03 ± 0.58 5.24 ± 0.68 4.91 ± 0.73 Inguinal fat 4.87 ± 1.54 4.40 ± 0.68 5.19 ± 0.90 Mesenteric fat 5.26 ± 1.01 4.77 ± 0.83 5.37 ± 0.95 Epididymal fat 0.88 ± 0.26 0.82 ± 0.23 0.68 ± 0.31 Data are means ± SE of 12 mice per group. p<0.05, p<0.01 vs WT mice. Table 3. Serum parameters in H-apoD Tg mice and WT littermates. WT Thy-1/ApoD NSE/ApoD Total protein (g/l) 52.20 ± 4.09 52.33 ± 5.16 55.40 ± 11.61 Albumin (g/l) 27.40 ±7.50 27.83 ± 6.71 30.20 ± 8.47 Glucose (mmol/l) 10.90 ± 0.75 12.52 ± 3.44 11.28 ± 2.95 Insulin (ng/ml) 1.44 ± 0.37 3.15 ± 0.56 2.32 ± 0.37 BUN urea (mmol/l) 8.28 ± 1.01 7.98 ± 1.79 7.62 ± 1.09 Creatinine ( mol/l) 13.33 ± 0.58 15.33 ± 5.13 17.67 ± 2.52 Total bilirubin ( mol/l) 2.92 ± 0.94 2.82 ± 1.48 3.56 ± 1.36 ALT (U/L) 39.00 ± 1.00 38.00 ± 2.65 72.00 ± 26.29 AST (U/L) 86.00 ± 20.88 78.67 ± 2.08 173.67 ± 65.26 Alkaline phosphatase (U/L) 82.40 ± 14.96 120.80 ± 62.50 73.00 ± 29.51 Creatine kinase (U/L) 85.33 ± 47.51 102.67 ± 27.01 94.00 ± 49.12 Cholesterol (mmol/l) 2.98 ± 0.36 3.13 ± 0.63 4.25 ± 1.98 HDL (mmol/l) 2.42 ± 0.31 2.60 ± 0.63 3.07 ± 1.57 Triglycerides (mmol/l) 1.15 ± 0.31 1.71 ± 0.89 1.14 ± 0.50 Data are means ± SE of 6 mice per group. p<0.05, p<0.01, p<0.001 vs WT mice. Do Carmo et al, AJP Endo Met 2008
TgApoD mice and hepatic steatosis Do Carmo et al, AJP Endo Met 2008
TgApoD mice characterization Do Carmo et al, AJP Endo Met 2008
TgApoD mice characterization 2.0 1.5 1.0 0.5 wt apod 3 2 1 0.0 PPARγ1 PPARγ2 pparg1 pparg2 wt tg PPARγ1 0 PPARγ2 PPARγ1 Amidoblack WT WT PPARγ mrna PPARγ protein PPARγ2 Liver Muscle HPRT C/EBPα mrna C/EBPβ mrna C/EBPα C/EBPβ HPRT HPRT Labrie et al. 2014 BBA-lipids submitted
TgApoD mice Lipid droplets formation 3 2 1 Plin2 protein (Relative to WT) Cide A mrna Cide B mrna (Relative to WT) 0 wt apod Plin2 Cide A Amidoblack HPRT Cide B HPRT Cide C mrna Lipid droplet size Cide C Lipid droplet number HPRT Labrie et al. 2014 BBA-lipids submitted
TgApoD mice and Fatty acid uptake 80000 60000 60000 40000 CD36 mrna CPM 3 H-oleate e / mg protein CPM 3 H-oleate/mg protein CD36 20000 0 WT Tg HPRT Labrie et al. 2014 BBA-lipids submitted
Hepatic lipogenesis Malonyl-CoA ACC Acetyl-CoA glycolysis Glucose FAS Palmitoyl-CoA Palmitoleyl-CoA C16:0 SCD1 C16:1 Stearoyl-CoA C18:0 Oleyl-CoA C18:1 Triglycerides Phospholipides Plasma Adipose tissues VLDL SFA MUFA Lipids deposits
TgApoD mice and lipogenesis AMPKα protein Phospho-AMPKα Expression AMPKα P-AMPKα Amidoblack Amidoblack ACC protein Phospho-ACC P-ACC/ ACC ACC Amidoblack P-ACC Amidoblack Labrie et al. 2014 BBA-lipids submitted
FAS protein FAS Amidoblack TgApoD mice and lipogenesis ACC Scd1 Dgat LXR-α HPRT WT mrna 0.16 0,16 0.14 0.12 0.10 0.08 0.06 0.04 0.02 0,14 0,12 0,10 0,08 0,06 0,04 0,02 ACC Scd1 3 H2 O incorporated/mg protein Ratio of 3 H2O incorporated/g/h 0 0,00 ns Labrie et al. 2014 BBA-lipids submitted
TgApoD mice and β-oxydation Labrie et al. 2014 BBA-lipids submitted
ApoD in hepg2 cells BSA AA 10 NT EV apod NT EV apod H-apoD β-actin Luciferase activity 8 6 4 2 apod AA(7µM) 0 - + - - - + + + - apod + apod PPRE PPRE PPRE LUC Myc β-actin Labrie et al. 2014 BBA-lipids submitted
ApoD mechanism of action FFA FFA FFA FFA FFA Lipid Droplet Cide-A Cide-C Plin2 TG FFA CD36 AA AA RXR Cide-C CD36 Plin2 Cide-A Labrie et al. 2014 BBA-lipids submitted
Acknowledgments Maryline Labrie Simon Lalonde Ouafa Nagyb Dr. E. Rassart