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Atherosclerotic biomarkers

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by

Ninna Lindberg

on 3 August 2013

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Transcript of Atherosclerotic biomarkers

Master thesis 2012/2013
Atherosclerotic biomarkers in transgenic Yucatan minipigs
Aim: to investigate whether a biomarker reflecting the activity of atherosclerosis or the degree of atherosclerosis development, can be found in transgenic Yucatan minipigs.
Project outline
ELISA: 4 candidate biomarkers
Proteomics: only proteins
Metabolomics: lipids, amino acids, carbohydrates and carbohydrate conjugates
ELISA
Proteomics
Metabolomics
a. iliaca communis
a. carotis communis
a. aorta abdominalis (1x1cm)
plasma
Sample collection
A. iliaca communis was divided in three; 0.1-0.2 g per 2 mL media
RPMI media (lipid, protein and growth factor-free)
24 hours
37 degrees C
5 % CO2
Incubation of vessels
Experimental animals
Lipoprotein-associated phospholipase A2 (Lp-PLA2)
soluble Intercellular adhesion molecule 1 (sICAM1)
High-sensitivity C-reactive protein
(hs-CRP)
A frequently discussed biomarker for atherosclerosis
acute phase reactant
detection limit: 0.3 µg/L (hs-CRP: 0.3 mg/L)
P = 0.0041
P = 0.8813
R2 = 0.0014
inflammatory enzyme implicated in formation of rupture prone plaque
inhibition reduces coronary plaque development (Wilensky et al. 2008)
The PLAC TEST for heart attack and stroke
travels mainly with LDL
hydrolyzes oxidized phospholipids in LDL
up-regulation owing to endothelial dysfunction (Nakashima et al. 1998)
glycoprotein
expressed on endothelial cells and cells of the immune system
mediates cell-cell interactions
P = 0.5824
R2 = 0.0182
P = 0.1932
Plasminogen activator inhibitor 1
(PAI-1)
the primary inhibitor of plasminogen activation (Sjöland et al. 1999; Sakata et al. 2004)
promotes atherosclerosis by facilitating fibrin deposition within developing lesions?
inhibitor of tPA and uPA
mainly produced by endothelial cells
Methods
Results
Plasma: 5033 features
Different adducts (H+, Na+, ACN)
Results
Media: 422 proteins
Plasma: 228 proteins
Sum up
Up-regulated:
Apolipoprotein A-II, A-IV, B-100, C-II, C-III, D, E.
Fibrinogen
Epidermal fatty acid binding protein 5 (x 430)
Orosomucoid 1 (x 11)
Periostin (x 9)
Phosphatidylethanolamine binding protein 1 (x 3)
Lumican (x 6)
Peroxiredoxin 2
Cathepsin S + Z
Plastin 2
Down-regulated:
Interleukin 6 (x 4)
CRP (x 3)
Keratin 1, 5, 8, 13, 14
D374Y-PCSK9 transgenic pigs
46 weeks
High-fat, high cholesterol diet
P = 0.0004
R2 = 0.5751
P = 0.7743
P = 0.0897
R2 = 0.1516
P = 0.0669
R2 = 0.1744
P = 0.2065
R2 = 0.0871
P = 0.9292
Correlation with total aorta plaque
P = 0.0089
R2 = 0.5117
P = 0.0111
R2 = 0.4912
P = 0.0151
R2 = 0.4618
Pearson r = 0.9285
P < 0.0001
R2 = 0.8621
P = 0.0014
R2 = 0.8906
Full transcript