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Genetic correlations between circulating miRNAs and lipid profiles reveal novel biomarkers of CVD risk
Abstracts / Atherosclerosis 252 (2016) e197ee235
Results: Cholesterol and triglyceride levels were significantly elevated in E3L and E3L.GK+/- mice compared to GK+/- mice. Fasting glucose was significantly increased in E3L.GK+/- and GK+/- compared to E3L (1.5-fold, < 0.001). Atherosclerotic lesion size (mm2) was significantly increased by 2.2fold in E3L.GK+/- (mean 68,291±41,809) as compared to E3L (mean 31,107±29,078)(P¼0.038; 1-tailed T-test). E3L.GK+/- had similar levels of hepatic steatosis and fibrosis as E3L. Histological evaluation of the kidneys revealed only mild morphological changes such as matrix expansion and dilatation of tubules. No clear pancreatic phenotype alterations were noted. Conclusions: We conclude that the E3L.GK+/- mouse is a promising novel diet-induced animal model for investigation of the etiology of diabetic macrovascular complications and evaluation of drug treatment thereon.
EAS16-0848, EXPERIMENTAL CADIOVASCULAR MEDICINE II. PPARg ACTIVATION ATTENUATES OXIDATIVE STRESS AND IMPROVES CEREBRAL ARTERIES ENDOTHELIAL DYSFUNCTION IN AGING RAT THROUGH UP-REGULATING UCP2 P. Wang, G. Cai, F. Wang, P. Zhou, Q. Wang. The First Affiliated HospitalChengdu Medical College, Department of Cardiology, Chengdu, China Objectives: Increasing amounts of evidence implicate oxidative stress as having a pivotal role in age-related cerebrovascular dysfunction, which is an important risk factor for the development of cerebrovascular disease. Previous studies have shown that the activation of the expression of peroxisome proliferator-activated receptor gamma (PPAR-g) in vascular endothelial cells results in an improvement of vascular function. Pioglitazone, a well-known PPAR-g agonist, protects against oxidative stress in the rostral ventrolateral medulla by the upregulation of mitochondrial uncoupling protein 2 (UCP2).In this study, we sought to explore the effects and the underlying mechanisms of pioglitazone on age-related oxidative stress elevation and cerebrovascular dysfunction in aging rat cerebral arteries. Methods: A natural aging model of Sprague Dawley rats was constructed and used in these experiments. Results: One-month oral administration of pioglitazone (20 mg$kg1$d1) ameliorated the production of reactive oxygen species, promoted endothelial nitric oxide synthase phosphorylation and increased the nitric oxide available, thus improving endothelium-dependent relaxation in aging rat cerebral arteries. One-month pioglitazone administration also restored PPAR-g expression and increased the levels of UCP2 in aging rat cerebral arteries. Using in vitro studies, we demonstrated that pioglitazone attenuated reactive oxygen species levels in aging human umbilical vein endothelial cells through PPAR-g activation. Furthermore, we found that this occurs in an UCP2-dependent manner. Conclusions: Our study demonstrated that the activation of PPAR-g by pioglitazone protected against oxidative stress damage in aging cerebral arteries by upregulating UCP2. PPAR-g may be a new target in treating agerelated cerebrovascular dysfunction.
EAS16-0975, EXPERIMENTAL CADIOVASCULAR MEDICINE II. EFFECTS OF CHRONIC INFLAMMATION ON MEGAKARYOCYTE AND PLATELET FUNCTION IN A CONDITIONAL MOUSE MODEL M. Salzmann, M. Haase, C. Arzt, M. Mussbacher, A. Assinger, J.A. €t Wien, Vascular Biology and Thrombosis Schmid. Medizinische Universita Research, Wien, Austria Objectives: The aim of this study was to determine the effect of inflammation on megakaryocytes and platelets. Megakaryocytes can sense inflammatory signals via TLRs but little is known how this might change platelets, which can contribute to cardiovascular diseases via immunemodulatory processes. Most inflammatory signalling pathways converge at the kinase IKK2 (inhibitor of NF-kB kinase 2) activating the transcription factor NF-kB. Our aim was to determine the effect of chronic inflammation on megakaryocytes and platelets by using conditional transgenic mouse models that alter NF-kB activity in megakaryocytes.
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Methods: Mice with a megakaryocyte-specific constitutively active IKK2 or IKK2 knock-out were compared to littermate controls, thereby studying different inflammatory states in megakaryocytes. Megakaryocytes were analysed with respect to their ploidy and ability to form pro-platelets. Blood platelets were analysed for size, number and granule content. Moreover, platelet functions were determined in vitro by testing for agonist-induced degranulation and aggregation. Results: We could not observe significant differences in megakaryocyte maturation and differentiation. Platelets of mice with megakaryocytespecific constitutive active IKK2 though, showed an increased volume and contained more RNA as compared to controls. Basal surface P-selectin expression was increased indicating constitutive degranulation, whereas platelet aggregation capability in vitro was significantly decreased. Conclusions: Our results suggest that platelets from megakaryocytes under persistent inflammatory stimulation have a higher basal activation, while reactivity is decreased. This may imply an “exhausted” phenotype and might represent a feedback mechanism that reduces thrombotic tendencies in states of chronic inflammation.
EAS16-0463, EXPERIMENTAL CADIOVASCULAR MEDICINE II. GENETIC CORRELATIONS BETWEEN CIRCULATING MIRNAS AND LIPID PROFILES REVEAL NOVEL BIOMARKERS OF CVD RISK L. Michael 1, S. McAhren 2, S. Kumar 3, J. Peralta 3, H. Kulkarni 3, W. Gerard 4, J. Weir 4, C. Barlow 4, M. Kowala 1, P. Meikle 4, J. Blangero 3, J. Curran 3. 1 Eli Lilly and Company, Cardiometabolic and Diabetes Complications, Indianapolis, USA; 2 Eli Lilly and Company, Bioinformatics, Indianapolis, USA; 3 University of Texas Rio Grande Valley School of Medicine, South Texas Diabetes and Obesity Institute, Brownsville, USA; 4 Baker IDI Heart and Diabetes Institute, Metabolomics, Melbourne, Australia Objectives: Cardiovascular disease (CVD) is the leading cause of death in the US, and prevention of CVD focuses on improving the lipid profile of patients at risk. The human plasma lipidome consists of thousands of lipid species. We have previously measured 315 such lipid species in the San Antonio Family Heart Study (SAFHS) cohort. These lipid species represent valuable endophenotypes for identifying genes involved in lipid metabolism related to CVD. Recent evidence has implicated microRNAs (miRNAs) in CVD pathogenesis. An expanded understanding of the function of miRNAs in gene networks associated with CVD will enable identification of novel mechanisms and biomarkers of disease. We conducted an analysis of circulating miRNAs associated with the human lipidome. Methods: We profiled plasma miRNAs in 384 SAFHS members to identify miRNAs that potentially influence the human lipidome as biomarkers or mediators of CVD. Results: Our analysis identified 402 known and 83 novel human miRNAs. The most striking association was observed between 4 different miRNAs and LPC 20:1: miR-330-3p (p¼1.21x10-7), miR-125b-5p (p¼4.61x10-7), miR-146a-5p (p¼2.18x10-6) and miR-130b-5p (p¼3.41x10-6). Additionally, miR-330-3p showed a significant association with LPC 16:1 (p¼1.48x10-6). All of these miRNAs were strongly correlated negatively with the lipid species indicating that increased lipid levels are associated with decreased miRNAs. LPC species have been shown previously to have favorable health effects including with decreased risk for CVD, decreased cIMT and HbA1c levels. Conclusions: Given the significant associations between these miRNAs and LPC species, our preliminary analyses indicate that these miRNAs may serve as biomarkers of increased genetic liability to CVD.
EAS16-0637, EXPERIMENTAL CADIOVASCULAR MEDICINE II. ELEVATED LDL AND OXIDATIVE STRESS CONTRIBUTE TO ARRHYTHMOGENIC CARDIOMYOPATHY PHENOTYPIC MANIFESTATION L. Arnaboldi 1, I. Stadiotti 2, S. Brambilla 2, G. Milano 2, A. Scopece 2, F. Cattaneo 3, A. Granata 1, C. Tondo 3, A. Corsini 1, G. Pompilio 2, E. degli Studi di Milano, Dipartimento di Scienze Sommariva 2. 1 Universita Farmacologiche e Biomolecolari, Milano, Italy; 2 Centro Cardiologico Monzino IRCCS, Vascular Biology and Regenerative Medicine Unit, Milano,
Results: Cholesterol and triglyceride levels were significantly elevated in E3L and E3L.GK+/- mice compared to GK+/- mice. Fasting glucose was significantly increased in E3L.GK+/- and GK+/- compared to E3L (1.5-fold, < 0.001). Atherosclerotic lesion size (mm2) was significantly increased by 2.2fold in E3L.GK+/- (mean 68,291±41,809) as compared to E3L (mean 31,107±29,078)(P¼0.038; 1-tailed T-test). E3L.GK+/- had similar levels of hepatic steatosis and fibrosis as E3L. Histological evaluation of the kidneys revealed only mild morphological changes such as matrix expansion and dilatation of tubules. No clear pancreatic phenotype alterations were noted. Conclusions: We conclude that the E3L.GK+/- mouse is a promising novel diet-induced animal model for investigation of the etiology of diabetic macrovascular complications and evaluation of drug treatment thereon.
EAS16-0848, EXPERIMENTAL CADIOVASCULAR MEDICINE II. PPARg ACTIVATION ATTENUATES OXIDATIVE STRESS AND IMPROVES CEREBRAL ARTERIES ENDOTHELIAL DYSFUNCTION IN AGING RAT THROUGH UP-REGULATING UCP2 P. Wang, G. Cai, F. Wang, P. Zhou, Q. Wang. The First Affiliated HospitalChengdu Medical College, Department of Cardiology, Chengdu, China Objectives: Increasing amounts of evidence implicate oxidative stress as having a pivotal role in age-related cerebrovascular dysfunction, which is an important risk factor for the development of cerebrovascular disease. Previous studies have shown that the activation of the expression of peroxisome proliferator-activated receptor gamma (PPAR-g) in vascular endothelial cells results in an improvement of vascular function. Pioglitazone, a well-known PPAR-g agonist, protects against oxidative stress in the rostral ventrolateral medulla by the upregulation of mitochondrial uncoupling protein 2 (UCP2).In this study, we sought to explore the effects and the underlying mechanisms of pioglitazone on age-related oxidative stress elevation and cerebrovascular dysfunction in aging rat cerebral arteries. Methods: A natural aging model of Sprague Dawley rats was constructed and used in these experiments. Results: One-month oral administration of pioglitazone (20 mg$kg1$d1) ameliorated the production of reactive oxygen species, promoted endothelial nitric oxide synthase phosphorylation and increased the nitric oxide available, thus improving endothelium-dependent relaxation in aging rat cerebral arteries. One-month pioglitazone administration also restored PPAR-g expression and increased the levels of UCP2 in aging rat cerebral arteries. Using in vitro studies, we demonstrated that pioglitazone attenuated reactive oxygen species levels in aging human umbilical vein endothelial cells through PPAR-g activation. Furthermore, we found that this occurs in an UCP2-dependent manner. Conclusions: Our study demonstrated that the activation of PPAR-g by pioglitazone protected against oxidative stress damage in aging cerebral arteries by upregulating UCP2. PPAR-g may be a new target in treating agerelated cerebrovascular dysfunction.
EAS16-0975, EXPERIMENTAL CADIOVASCULAR MEDICINE II. EFFECTS OF CHRONIC INFLAMMATION ON MEGAKARYOCYTE AND PLATELET FUNCTION IN A CONDITIONAL MOUSE MODEL M. Salzmann, M. Haase, C. Arzt, M. Mussbacher, A. Assinger, J.A. €t Wien, Vascular Biology and Thrombosis Schmid. Medizinische Universita Research, Wien, Austria Objectives: The aim of this study was to determine the effect of inflammation on megakaryocytes and platelets. Megakaryocytes can sense inflammatory signals via TLRs but little is known how this might change platelets, which can contribute to cardiovascular diseases via immunemodulatory processes. Most inflammatory signalling pathways converge at the kinase IKK2 (inhibitor of NF-kB kinase 2) activating the transcription factor NF-kB. Our aim was to determine the effect of chronic inflammation on megakaryocytes and platelets by using conditional transgenic mouse models that alter NF-kB activity in megakaryocytes.
e227
Methods: Mice with a megakaryocyte-specific constitutively active IKK2 or IKK2 knock-out were compared to littermate controls, thereby studying different inflammatory states in megakaryocytes. Megakaryocytes were analysed with respect to their ploidy and ability to form pro-platelets. Blood platelets were analysed for size, number and granule content. Moreover, platelet functions were determined in vitro by testing for agonist-induced degranulation and aggregation. Results: We could not observe significant differences in megakaryocyte maturation and differentiation. Platelets of mice with megakaryocytespecific constitutive active IKK2 though, showed an increased volume and contained more RNA as compared to controls. Basal surface P-selectin expression was increased indicating constitutive degranulation, whereas platelet aggregation capability in vitro was significantly decreased. Conclusions: Our results suggest that platelets from megakaryocytes under persistent inflammatory stimulation have a higher basal activation, while reactivity is decreased. This may imply an “exhausted” phenotype and might represent a feedback mechanism that reduces thrombotic tendencies in states of chronic inflammation.
EAS16-0463, EXPERIMENTAL CADIOVASCULAR MEDICINE II. GENETIC CORRELATIONS BETWEEN CIRCULATING MIRNAS AND LIPID PROFILES REVEAL NOVEL BIOMARKERS OF CVD RISK L. Michael 1, S. McAhren 2, S. Kumar 3, J. Peralta 3, H. Kulkarni 3, W. Gerard 4, J. Weir 4, C. Barlow 4, M. Kowala 1, P. Meikle 4, J. Blangero 3, J. Curran 3. 1 Eli Lilly and Company, Cardiometabolic and Diabetes Complications, Indianapolis, USA; 2 Eli Lilly and Company, Bioinformatics, Indianapolis, USA; 3 University of Texas Rio Grande Valley School of Medicine, South Texas Diabetes and Obesity Institute, Brownsville, USA; 4 Baker IDI Heart and Diabetes Institute, Metabolomics, Melbourne, Australia Objectives: Cardiovascular disease (CVD) is the leading cause of death in the US, and prevention of CVD focuses on improving the lipid profile of patients at risk. The human plasma lipidome consists of thousands of lipid species. We have previously measured 315 such lipid species in the San Antonio Family Heart Study (SAFHS) cohort. These lipid species represent valuable endophenotypes for identifying genes involved in lipid metabolism related to CVD. Recent evidence has implicated microRNAs (miRNAs) in CVD pathogenesis. An expanded understanding of the function of miRNAs in gene networks associated with CVD will enable identification of novel mechanisms and biomarkers of disease. We conducted an analysis of circulating miRNAs associated with the human lipidome. Methods: We profiled plasma miRNAs in 384 SAFHS members to identify miRNAs that potentially influence the human lipidome as biomarkers or mediators of CVD. Results: Our analysis identified 402 known and 83 novel human miRNAs. The most striking association was observed between 4 different miRNAs and LPC 20:1: miR-330-3p (p¼1.21x10-7), miR-125b-5p (p¼4.61x10-7), miR-146a-5p (p¼2.18x10-6) and miR-130b-5p (p¼3.41x10-6). Additionally, miR-330-3p showed a significant association with LPC 16:1 (p¼1.48x10-6). All of these miRNAs were strongly correlated negatively with the lipid species indicating that increased lipid levels are associated with decreased miRNAs. LPC species have been shown previously to have favorable health effects including with decreased risk for CVD, decreased cIMT and HbA1c levels. Conclusions: Given the significant associations between these miRNAs and LPC species, our preliminary analyses indicate that these miRNAs may serve as biomarkers of increased genetic liability to CVD.
EAS16-0637, EXPERIMENTAL CADIOVASCULAR MEDICINE II. ELEVATED LDL AND OXIDATIVE STRESS CONTRIBUTE TO ARRHYTHMOGENIC CARDIOMYOPATHY PHENOTYPIC MANIFESTATION L. Arnaboldi 1, I. Stadiotti 2, S. Brambilla 2, G. Milano 2, A. Scopece 2, F. Cattaneo 3, A. Granata 1, C. Tondo 3, A. Corsini 1, G. Pompilio 2, E. degli Studi di Milano, Dipartimento di Scienze Sommariva 2. 1 Universita Farmacologiche e Biomolecolari, Milano, Italy; 2 Centro Cardiologico Monzino IRCCS, Vascular Biology and Regenerative Medicine Unit, Milano,