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Effect of six type 2 diabetes susceptibility loci and the FTO variant RS9939609 on obesity in Pakistani obese subjects
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Abstracts / Atherosclerosis 237 (2014) e1ee16
immunocytochemistry for BrdU/EdU incorporation, and cleaved caspase3, respectively. Wnt4 significantly increased VSMC proliferation (1.87±0.16 fold vs control, n¼3, p<0.05), whilst Wnt3a, Wnt5a and Wnt9a had no effect. Conversely, recombinant Wnt3a, Wnt5a and Wnt9a proteins significantly suppressed H₂O₂-induced apoptosis (by 60±14%, 66±17%, 43±12% respectively, n¼3, p<0.05), but Wnt4 had no effect. We demonstrated by quantitative-PCR that Wnt3a, Wnt5a, and Wnt9a, but not Wnt4, significantly increased expression of the pro-survival factor, WISP-1 (1.9±0.2, 2.9±0.4, 1.7±0.3 fold vs control, n¼6, p<0.05). Wnt-induced survival was CREB- and WISP-1-dependent and overexpression of a dominant negative form of CREB (RAd-A-CREB) significantly inhibited Wnt-induced WISP-1 expression (0.6±0.1 fold vs RAd-b-galactosidase control, n¼3, p<0.05). Our data suggests that Wnt3a, Wnt4, Wnt5a and Wnt9a are expressed in atherosclerotic plaques within the vicinity of VSMCs. However, the effects of these Wnts on VSMCs are divergent: Wnt4 promotes VSMC proliferation, while Wnt3a, Wnt5a and Wnt9a protect from apoptosis. Consequently, these Wnts may have differing effects on atherosclerosis. FORECASTING VULNERABLE AEMORHEOLOGICAL FLOW
PLAQUE
RESPONSE
TO
G. Pereira*, R. Krams. Bioengineering Department Imperial College London, South Kensington Campus, 4.28 Royal School of Mines, UK
* Corresponding author. Research related to Atherosclerosis and the mechanisms by which plaque initiates and develops is the focus of major studies on healthcare [1]. It is found to be directly and/or indirectly related to a burden of diseases. Nowadays, experimental and theoretical/computational findings have been used to clarify some mechanisms behind atherosclerosis [2,3,4]. However, there is a number of phenomena that is not totally explained on current literature, as for instance, the relation between the non-Newtonian blood flow and metabolic processes that drive atheroma formation. The focus of this research is on how aemorheology correlates with vulnerable plaque phenotype. To understand the coupling between mechanical and biophysical mechanisms, we are simulating non-newtonian blood flow and analysing differences on flow patterns due to blood particles. The geometries of interest are based on stenosis and arterial branches. The former is based on image reconstructions of an Apoe+/- mice model of atherosclerosis [5]. Initial studies provide information about the influence of modelling and computational set up, including mesh characteristics, on results. We report on differences ranging from 10 to 48% in flow patterns due to model and computational definitions. The results are comparable to literature (about 90% of accuracy) [6,7]. Currently, we are correlating nonNewtonian flow patterns with plaque phenotype. [1] [2] [3] [4] [5] [6] [7]
http://www.bhf.org.uk/ C. G. Caro et al., Nature, 1969. C. Chen et al.. Biorheology, 2011. S. Sapna, I.J.Phys.Sciences, 2009. V. Mehta, PhD Thesis, ongoing. H. Huang et. al., Int.J.Engng. Sci., 1995. J. M. Reese and D. S. Thompson, J.Biomech.,1998.
SYSTEMS BIOLOGY MODELS TO TEST PHYSIOLOGICAL AND PHARMACOLOGICAL HYPOTHESIS IN THE STUDY OF ATHEROSCLEROSIS Cesar Pichardo-Almarza 1, 2, *, Vanessa Diaz-Zuccarini 1. 1 Multiscale Cardiovascular Engineering Group, Department of Mechanical Engineering, University College London, UK; 2 Xenologiq Ltd, Canterbury, UK
* Corresponding author. Rationale for the study: The systematic analysis and comprehensive and quantitative understanding of atherosclerosis formation is becoming essential, now that patients are consistently treated with statins, even if there are controversies over their benefit/detriment ratio. Mathematical models can be used to shed light on new mechanisms and to provide a
better description of the physiological effects and the pharmacokinetic properties of a given drug. Here, we show how a ‘systems approach’ is used to develop a new model of atherosclerosis where mechanisms from different physical domains and biological scales can be combined with the pharmacological properties of a given drug (i.e. statins). The iteration between computational (and/or mathematical) modelling and experimentation provides a holistic description of the effect of statins in atherosclerosis progression. Methodology and results: The purpose of this model is to better understand the dynamics of inflammatory mechanisms related to early atherosclerotic lesions, by studying the influence of lipoproteins, flowrelated variables and the mechanisms of cholesterol-lowering drugs on plaque progression. Published data from intravascular ultrasound studies was used for validation purposes. The novel work presented here shows an integrative systems model, which simulates the dynamics of a (given) statin and its effect on atherosclerosis formation. Conclusions: A new system model explaining relationships between lipoprotein dynamics and statins on early atherosclerotic lesions was developed, providing a description of relevant inflammatory processes. This model describes dynamically the development of atherosclerosis and could potentially incorporate additional factors needed for a better understanding of the disease and its treatment. Acknowledgments This work was supported by the EPSRC grant “Personalised Medicine Through Learning in the Model Space” (grant number EP/L000296/1). EFFECT OF SIX TYPE 2 DIABETES SUSCEPTIBILITY LOCI AND THE FTO VARIANT RS9939609 ON OBESITY IN PAKISTANI OBESE SUBJECTS Shabana 1, *, Ka Wah Li 2, Jayshree Acharya 2, Jackie A. Cooper 2, Shahida Hasnain 1, Stephen E. Humphries 2. 1 Department of Microbiology and Molecular Genetics, University of the Punjab, Lahore 54590, Pakistan; 2 Centre for Cardiovascular Genetics, British Heart Foundation Laboratories, The Rayne Building, Institute of Cardiovascular Sciences, University College London, London WC1E 6JF, UK
* Corresponding author. Objectives: We aimed to analyze effect of six type 2 diabetes susceptibility loci, rs3923113 (GRB14), rs18681329 (ST6GAL1), rs1802295 (VPS26A), rs7178572 (HMG20A), rs2028299 (AP3S2), and rs4812829 (HNF4A) and FTO polymorphism (rs9939609) reported to have contradictory results in Caucasians and Asians, on obesity. Association between the SNPs and biochemical and anthropometric parameters was analyzed to deduce mechanism of action. Design and methods: 475 samples were genotyped by TaqMan assay and lipid profile was determined. Allele & genotype frequencies and genescore were calculated. The effect of genescore on anthropometric and biochemical parameters was analyzed. Results: All variants had minor allele frequency comparable to the original GWAS and were associated with obesity in Pakistani subjects. A threshold effect of having at least 5 risk alleles to affect body weight considerably was observed. The obesity genescore was significantly associated with increasing cholesterol (p¼1.3510-13), triglycerides (p¼1.0510-5), LDL-C (p¼8.7310-3) and decreasing HDL-C (p¼0.002). We are first to report association of GWAS SNPs with obesity. Conclusion: We confirm a similar effect of SNPs for obesity as in the original study in Pakistani population. Effect of having a combination of risk alleles on obesity is considerable. The mechanism of this effect may be mediated by changing serum lipid chemistry. EFFECT OF SORTI, APOB AND APOE POLYMORPHISMS ON LDL-C IN BRITISH AND PAKISTANI PEOPLE AND THEIR ASSOCIATION WITH RISK OF CAD IN PAKISTANI POPULATION Saleem Ullah Shahid 1, *, Jackie A. Cooper 2, Abdul Rehman 1, Stephen E. Humphries 2. 1 Department of Microbiology and Molecular Genetics, University of the Punjab, Lahore 54590, Pakistan; 2 Centre for
Abstracts / Atherosclerosis 237 (2014) e1ee16
immunocytochemistry for BrdU/EdU incorporation, and cleaved caspase3, respectively. Wnt4 significantly increased VSMC proliferation (1.87±0.16 fold vs control, n¼3, p<0.05), whilst Wnt3a, Wnt5a and Wnt9a had no effect. Conversely, recombinant Wnt3a, Wnt5a and Wnt9a proteins significantly suppressed H₂O₂-induced apoptosis (by 60±14%, 66±17%, 43±12% respectively, n¼3, p<0.05), but Wnt4 had no effect. We demonstrated by quantitative-PCR that Wnt3a, Wnt5a, and Wnt9a, but not Wnt4, significantly increased expression of the pro-survival factor, WISP-1 (1.9±0.2, 2.9±0.4, 1.7±0.3 fold vs control, n¼6, p<0.05). Wnt-induced survival was CREB- and WISP-1-dependent and overexpression of a dominant negative form of CREB (RAd-A-CREB) significantly inhibited Wnt-induced WISP-1 expression (0.6±0.1 fold vs RAd-b-galactosidase control, n¼3, p<0.05). Our data suggests that Wnt3a, Wnt4, Wnt5a and Wnt9a are expressed in atherosclerotic plaques within the vicinity of VSMCs. However, the effects of these Wnts on VSMCs are divergent: Wnt4 promotes VSMC proliferation, while Wnt3a, Wnt5a and Wnt9a protect from apoptosis. Consequently, these Wnts may have differing effects on atherosclerosis. FORECASTING VULNERABLE AEMORHEOLOGICAL FLOW
PLAQUE
RESPONSE
TO
G. Pereira*, R. Krams. Bioengineering Department Imperial College London, South Kensington Campus, 4.28 Royal School of Mines, UK
* Corresponding author. Research related to Atherosclerosis and the mechanisms by which plaque initiates and develops is the focus of major studies on healthcare [1]. It is found to be directly and/or indirectly related to a burden of diseases. Nowadays, experimental and theoretical/computational findings have been used to clarify some mechanisms behind atherosclerosis [2,3,4]. However, there is a number of phenomena that is not totally explained on current literature, as for instance, the relation between the non-Newtonian blood flow and metabolic processes that drive atheroma formation. The focus of this research is on how aemorheology correlates with vulnerable plaque phenotype. To understand the coupling between mechanical and biophysical mechanisms, we are simulating non-newtonian blood flow and analysing differences on flow patterns due to blood particles. The geometries of interest are based on stenosis and arterial branches. The former is based on image reconstructions of an Apoe+/- mice model of atherosclerosis [5]. Initial studies provide information about the influence of modelling and computational set up, including mesh characteristics, on results. We report on differences ranging from 10 to 48% in flow patterns due to model and computational definitions. The results are comparable to literature (about 90% of accuracy) [6,7]. Currently, we are correlating nonNewtonian flow patterns with plaque phenotype. [1] [2] [3] [4] [5] [6] [7]
http://www.bhf.org.uk/ C. G. Caro et al., Nature, 1969. C. Chen et al.. Biorheology, 2011. S. Sapna, I.J.Phys.Sciences, 2009. V. Mehta, PhD Thesis, ongoing. H. Huang et. al., Int.J.Engng. Sci., 1995. J. M. Reese and D. S. Thompson, J.Biomech.,1998.
SYSTEMS BIOLOGY MODELS TO TEST PHYSIOLOGICAL AND PHARMACOLOGICAL HYPOTHESIS IN THE STUDY OF ATHEROSCLEROSIS Cesar Pichardo-Almarza 1, 2, *, Vanessa Diaz-Zuccarini 1. 1 Multiscale Cardiovascular Engineering Group, Department of Mechanical Engineering, University College London, UK; 2 Xenologiq Ltd, Canterbury, UK
* Corresponding author. Rationale for the study: The systematic analysis and comprehensive and quantitative understanding of atherosclerosis formation is becoming essential, now that patients are consistently treated with statins, even if there are controversies over their benefit/detriment ratio. Mathematical models can be used to shed light on new mechanisms and to provide a
better description of the physiological effects and the pharmacokinetic properties of a given drug. Here, we show how a ‘systems approach’ is used to develop a new model of atherosclerosis where mechanisms from different physical domains and biological scales can be combined with the pharmacological properties of a given drug (i.e. statins). The iteration between computational (and/or mathematical) modelling and experimentation provides a holistic description of the effect of statins in atherosclerosis progression. Methodology and results: The purpose of this model is to better understand the dynamics of inflammatory mechanisms related to early atherosclerotic lesions, by studying the influence of lipoproteins, flowrelated variables and the mechanisms of cholesterol-lowering drugs on plaque progression. Published data from intravascular ultrasound studies was used for validation purposes. The novel work presented here shows an integrative systems model, which simulates the dynamics of a (given) statin and its effect on atherosclerosis formation. Conclusions: A new system model explaining relationships between lipoprotein dynamics and statins on early atherosclerotic lesions was developed, providing a description of relevant inflammatory processes. This model describes dynamically the development of atherosclerosis and could potentially incorporate additional factors needed for a better understanding of the disease and its treatment. Acknowledgments This work was supported by the EPSRC grant “Personalised Medicine Through Learning in the Model Space” (grant number EP/L000296/1). EFFECT OF SIX TYPE 2 DIABETES SUSCEPTIBILITY LOCI AND THE FTO VARIANT RS9939609 ON OBESITY IN PAKISTANI OBESE SUBJECTS Shabana 1, *, Ka Wah Li 2, Jayshree Acharya 2, Jackie A. Cooper 2, Shahida Hasnain 1, Stephen E. Humphries 2. 1 Department of Microbiology and Molecular Genetics, University of the Punjab, Lahore 54590, Pakistan; 2 Centre for Cardiovascular Genetics, British Heart Foundation Laboratories, The Rayne Building, Institute of Cardiovascular Sciences, University College London, London WC1E 6JF, UK
* Corresponding author. Objectives: We aimed to analyze effect of six type 2 diabetes susceptibility loci, rs3923113 (GRB14), rs18681329 (ST6GAL1), rs1802295 (VPS26A), rs7178572 (HMG20A), rs2028299 (AP3S2), and rs4812829 (HNF4A) and FTO polymorphism (rs9939609) reported to have contradictory results in Caucasians and Asians, on obesity. Association between the SNPs and biochemical and anthropometric parameters was analyzed to deduce mechanism of action. Design and methods: 475 samples were genotyped by TaqMan assay and lipid profile was determined. Allele & genotype frequencies and genescore were calculated. The effect of genescore on anthropometric and biochemical parameters was analyzed. Results: All variants had minor allele frequency comparable to the original GWAS and were associated with obesity in Pakistani subjects. A threshold effect of having at least 5 risk alleles to affect body weight considerably was observed. The obesity genescore was significantly associated with increasing cholesterol (p¼1.3510-13), triglycerides (p¼1.0510-5), LDL-C (p¼8.7310-3) and decreasing HDL-C (p¼0.002). We are first to report association of GWAS SNPs with obesity. Conclusion: We confirm a similar effect of SNPs for obesity as in the original study in Pakistani population. Effect of having a combination of risk alleles on obesity is considerable. The mechanism of this effect may be mediated by changing serum lipid chemistry. EFFECT OF SORTI, APOB AND APOE POLYMORPHISMS ON LDL-C IN BRITISH AND PAKISTANI PEOPLE AND THEIR ASSOCIATION WITH RISK OF CAD IN PAKISTANI POPULATION Saleem Ullah Shahid 1, *, Jackie A. Cooper 2, Abdul Rehman 1, Stephen E. Humphries 2. 1 Department of Microbiology and Molecular Genetics, University of the Punjab, Lahore 54590, Pakistan; 2 Centre for