RESUMEN
The PLPP3 gene encodes for a ubiquitous enzyme that dephosphorylates several lipid substrates. Genome-wide association studies identified PLPP3 as a gene that plays a role in coronary artery disease susceptibility. The aim of the study was to investigate the effect of Plpp3 deletion on atherosclerosis development in mice. Because the constitutive deletion of Plpp3 in mice is lethal, conditional Plpp3 hepatocyte-specific null mice were generated by crossing floxed Plpp3 mice with animals expressing Cre recombinase under control of the albumin promoter. The mice were crossed onto the athero-prone apoE-/- background to obtain Plpp3f/fapoE-/-Alb-Cre+ and Plpp3f/fapoE-/-Alb-Cre- offspring, the latter of which were used as controls. The mice were fed chow or a Western diet for 32 or 12 weeks, respectively. On the Western diet, Alb-Cre+ mice developed more atherosclerosis than Alb-Cre- mice, both at the aortic sinus and aorta. Lipidomic analysis showed that hepatic Plpp3 deletion significantly modified the levels of several plasma lipids involved in atherosclerosis, including lactosylceramides, lysophosphatidic acids, and lysophosphatidylinositols. In conclusion, Plpp3 ablation in mice worsened atherosclerosis development. Lipidomic analysis suggested that the hepatic Plpp3 deletion may promote atherosclerosis by increasing plasma levels of several low-abundant pro-atherogenic lipids, thus providing a molecular basis for the observed results.
Asunto(s)
Apolipoproteínas E/genética , Aterosclerosis/genética , Hígado/metabolismo , Fosfatidato Fosfatasa/genética , Animales , Aorta/metabolismo , Aorta/patología , Aterosclerosis/sangre , Aterosclerosis/patología , Dieta Occidental/efectos adversos , Modelos Animales de Enfermedad , Eliminación de Gen , Hepatocitos/metabolismo , Hepatocitos/patología , Humanos , Metabolismo de los Lípidos/genética , Lípidos/sangre , Ratones , Ratones Noqueados , Especificidad de Órganos/genéticaRESUMEN
OBJECTIVES: Inhibition of proprotein convertase subtilisin/kexin type 9 (PCSK9) has been proposed to be a potential new therapeutic target for treatment of hypercholesterolaemia. However, little is known about the effects of PCSK9 inhibition on the lipidome. METHODS: We performed molecular lipidomic analyses of plasma samples obtained from PCSK9-deficient mice, and serum of human carriers of a loss-of-function variant in the PCSK9 gene (R46L). RESULTS: In both mouse and man, PCSK9 deficiency caused a decrease in several cholesteryl esters (CE) and short fatty acid chain containing sphingolipid species such as CE 16:0, glucosyl/galactosylceramide (Glc/GalCer) d18:1/16:0, and lactosylceramide (LacCer) d18:1/16:0. In mice, the changes in lipid concentrations were most prominent when animals were given regular chow diet. In man, a number of molecular lipid species was shown to decrease significantly even when LDL-cholesterol was non-significantly reduced by 10% only. Western diet attenuated the lipid lowering potency of PCSK9 deficiency in mice. CONCLUSIONS: Plasma molecular lipid species may be utilized for characterizing novel compounds inhibiting PCSK9 and as sensitive efficacy markers of the PCSK9 inhibition.
Asunto(s)
LDL-Colesterol/sangre , Proproteína Convertasas/deficiencia , Serina Endopeptidasas/deficiencia , Esfingolípidos/sangre , Animales , Biomarcadores/sangre , Ésteres del Colesterol/sangre , Genotipo , Humanos , Masculino , Ratones , Ratones Noqueados , Mutación , Fenotipo , Proproteína Convertasa 9 , Proproteína Convertasas/genética , Estudios Prospectivos , Serina Endopeptidasas/genéticaRESUMEN
OBJECTIVE: Although relationships between smoking and cardiovascular diseases (CVD), and between CVD and lipids are established, the direct impact of smoking on lipidomes is not well understood. We investigated the effect of mainstream cigarette smoke (CS) exposure on plasma, liver, and aorta molecular lipid profiles, and liver transcriptome in the ApoE(-/-) mouse, a well-established mouse model for human atherogenesis. METHODS: Plasma, liver, and aorta samples from ApoE(-/-) mice exposed to CS or fresh air (sham) for six months were extracted for lipids using robotic-assisted method and analyzed by mass spectrometry. Gene expression in the liver was obtained on microarrays. Development of atherosclerosis in the aorta was further assessed by plaque size in the aortic arch and lipoprotein concentration in plasma and plaque. RESULTS: CS increased most lipid classes and molecular lipid species. In plasma, free cholesterol, ceramides, cerebrosides, and most phospholipids were increased in CS-exposed mice. In the liver, several lipid species including free and esterified cholesterol, triacylglycerols, phospholipids, sphingomyelins, and ceramides were elevated. In the aorta, more than 2-fold higher cholesteryl ester (CE), lysophosphatidylcholine, and glucosyl/galactosylceramide levels were seen. Moreover, CS exposure induced a significant decrease in several plasma CE and phosphatidylcholine species that contained polyunsaturated fatty acids. Genes involved in amino acid and lipid metabolism showed perturbed transcription profiles in the liver. CONCLUSION: We have quantified some of the molecular changes that accompany the increase of plaque size that is accelerated by CS exposure in the aortae of ApoE(-/-) mice. These results suggest that specific changes in the lipidome and transcriptome, for example in ceramide and polyunsaturated fatty acid species, may be associated with atherosclerosis.
Asunto(s)
Aorta Torácica/efectos de los fármacos , Enfermedades de la Aorta/sangre , Enfermedades de la Aorta/etiología , Apolipoproteínas E/deficiencia , Aterosclerosis/etiología , Lípidos/sangre , Hígado/efectos de los fármacos , Contaminación por Humo de Tabaco/efectos adversos , Animales , Aorta Torácica/metabolismo , Aorta Torácica/patología , Enfermedades de la Aorta/genética , Enfermedades de la Aorta/patología , Apolipoproteínas E/genética , Aterosclerosis/sangre , Aterosclerosis/genética , Aterosclerosis/patología , Modelos Animales de Enfermedad , Femenino , Perfilación de la Expresión Génica/métodos , Regulación de la Expresión Génica , Redes Reguladoras de Genes , Hígado/metabolismo , Espectrometría de Masas , Ratones , Ratones Noqueados , Análisis de Secuencia por Matrices de Oligonucleótidos , Placa Aterosclerótica , Factores de TiempoRESUMEN
Lipids, abundant constituents of both the vascular plaque and lipoproteins, play a pivotal role in atherosclerosis. Mass spectrometry-based analysis of lipids, called lipidomics, presents a number of opportunities not only for understanding the cellular processes in health and disease but also in enabling personalized medicine. Lipidomics in its most advanced form is able to quantify hundreds of different molecular lipid species with various structural and functional roles. Unraveling this complexity will improve our understanding of diseases such as atherosclerosis at a level of detail not attainable with classical analytical methods. Improved patient selection, biomarkers for gauging treatment efficacy and safety, and translational models will be facilitated by the lipidomic deliverables. Importantly, lipid-based biomarkers and targets should lead the way as we progress toward more specialized therapeutics.
Asunto(s)
Aterosclerosis/diagnóstico , Aterosclerosis/metabolismo , Lípidos/análisis , Aterosclerosis/tratamiento farmacológico , Biomarcadores/análisis , Humanos , Metabolismo de los Lípidos , Lipoproteínas/metabolismo , Espectrometría de Masas , Modelos AnimalesRESUMEN
BACKGROUND: The number of patients eligible for cardiovascular therapies in general is forecast to increase substantially in the coming decades. However, the current list of potential future cardiovascular blockbuster drugs is alarmingly short. There is thus a clear need for innovative strategies to increase the efficiency of drug development pipelines by establishing new sensitive biomarkers to monitor drug efficacy and safety in the context of complexity of lipoprotein metabolism targeted by the cardiovascular drugs. METHODS: Metabolomics is a discipline dedicated to the systematic study of small molecules in cells, tissues and biofluids. Since lipids (including cholesterol), as well as other metabolites, are key constituents of lipoprotein particles and are thus part of the complex lipoprotein metabolism that includes exchange of lipids and metabolites with peripheral tissues, cardiovascular drug safety and efficacy needs to be addressed in the context of systemic lipid metabolism. RESULTS/CONCLUSION: Metabolomics, lipidomics in particular, is expected to make an important impact on the discovery and development of cardiovascular therapies.
Asunto(s)
Fármacos Cardiovasculares/farmacología , Enfermedades Cardiovasculares/tratamiento farmacológico , Inhibidores de Hidroximetilglutaril-CoA Reductasas/farmacología , Metabolismo de los Lípidos/efectos de los fármacos , Animales , Biomarcadores Farmacológicos/metabolismo , Fármacos Cardiovasculares/efectos adversos , Enfermedades Cardiovasculares/metabolismo , Humanos , Inhibidores de Hidroximetilglutaril-CoA Reductasas/efectos adversos , Fenotipo , Resultado del TratamientoRESUMEN
Recent data suggest that aggressive lipid-lowering treatment results in significant reductions of atherosclerotic complications, ie, strokes, heart attacks, or peripheral vascular diseases. Thus, more patients will be titrated to higher doses of statins in order to reach the aggressive targets of low-density lipoprotein- cholesterol reduction. However, aggressive treatment with high statin doses has increased the risk of statin-induced myopathy. The incidence of myopathy in cohort studies and in randomized trials has been low, supporting the good safety profile of statin drugs. However, muscle effects seem to be more frequent in clinical practice. Of all statin users, approximately 1% to 5% suffers from muscular symptoms caused by medication. This potentially reduces the compliance toward treatment and number of patients reaching their treatment targets due to withdrawal of therapy. Thus, novel biomarkers are needed for prediction or improved diagnoses of statin-induced side effects. This would potentially increase the quality of life of patients and improve treatment results. Using lipidomic analysis, we found that the plasma lipidomic changes following simvastatin treatment correlate with the muscle expression of the arachidonate 5-lipoxygenase-activating protein. Intriguingly, these results suggest that the plasma lipidomic profile may serve as a highly sensitive biomarker of statin-induced metabolic alterations in muscle and may thus allow us to identify patients who should be treated with a lower dose to prevent a possible toxicity.
Asunto(s)
Biología Computacional , Inhibidores de Hidroximetilglutaril-CoA Reductasas/efectos adversos , Metabolismo de los Lípidos/fisiología , Enfermedades Musculares/inducido químicamente , Proteínas Activadoras de la 5-Lipooxigenasa , Biomarcadores/metabolismo , Proteínas Portadoras/metabolismo , Humanos , Proteínas de la Membrana/metabolismo , Músculo Esquelético/metabolismo , Enfermedades Musculares/metabolismoRESUMEN
Phospholipid transfer protein (PLTP) exists in a high-activity (HA-PLTP) and a low-activity form (LA-PLTP) in the circulation. LA-PLTP is associated with apoA-I while the HA-PLTP complex is enriched with apoE. To study the interaction of PLTP with apolipoproteins, we carried out surface plasmon resonance analyses. These demonstrated a concentration-dependent binding of recombinant human PLTP, which represents an active PLTP form, and LA-PLTP to apoE, apoA-I, and apoA-IV within a nanomolar K(D) range. To study whether LA-PLTP can be transformed into an active form, we incubated it in the presence of proteoliposomes containing apoE, apoA-I or apoA-IV. The apoE proteoliposomes induced a concentration-dependent activation of LA-PLTP. ApoA-IV proteoliposomes also activated LA-PLTP in a concentration-dependent manner, whereas apoA-I proteoliposomes had no such effect. These observations suggest that PLTP is capable of interacting with apoE, apoA-I, and apoA-IV, and that these interactions regulate PLTP-activity levels in plasma.
Asunto(s)
Apolipoproteínas E/metabolismo , Proteínas de la Membrana/metabolismo , Proteínas de Transferencia de Fosfolípidos/metabolismo , Apolipoproteína A-I/metabolismo , Apolipoproteínas A/metabolismo , Humanos , Proteolípidos/química , Resonancia por Plasmón de SuperficieRESUMEN
Human serum phospholipid transfer protein (PLTP) exists as a catalytically active (HA-PLTP) and a low-active (LA-PLTP) form. In this study, the association of PLTP activity and the concentrations of both forms with lipid and carbohydrate parameters were investigated. In a random Finnish population sample, serum PLTP concentration (n=250) was 6.56 +/- 1.45 mg/l, the mean lipoprotein-independent (PLTPexo) phospholipid transfer activity was 6.59 +/- 1.66 micromol/ml/h, and the mean lipoprotein-dependent (PLTPendo) activity was 1.37 +/- 0.29 micromol/ml/h. Of the serum PLTP concentration, approximately 46% was in a catalytically active form. HA-PLTP concentration correlated positively with serum PLTPexo activity (r=0.380, P <0.001), HDL cholesterol (r=0.291, P <0.001), and apolipoprotein A-I (r=0.187, P <0.01). Of the potential regulatory factors for PLTP, apolipoprotein E showed a weak positive correlation with serum PLTPexo (r=0.154, P <0.05) and PLTPendo (r=0.192, P <0.01) activity but not with PLTP concentration. Weak associations were also observed between PLTP parameters and determinants of glucose homeostasis (glucose, insulin, and homeostasis model assessment for insulin resistance). The present data on PLTP activity and concentration reveal novel connections of the two PLTP forms to lipid and carbohydrate metabolism.