RESUMEN
HDL cholesterol (HDL-C) remains a superior biochemical predictor of CVD risk, but its genetic basis is incompletely defined. In patients with extreme HDL-C concentrations, we concurrently evaluated the contributions of multiple large- and small-effect genetic variants. In a discovery cohort of 255 unrelated lipid clinic patients with extreme HDL-C levels, we used a targeted next-generation sequencing panel to evaluate rare variants in known HDL metabolism genes, simultaneously with common variants bundled into a polygenic trait score. Two additional cohorts were used for validation and included 1,746 individuals from the Montréal Heart Institute Biobank and 1,048 individuals from the University of Pennsylvania. Findings were consistent between cohorts: we found rare heterozygous large-effect variants in 18.7% and 10.9% of low- and high-HDL-C patients, respectively. We also found common variant accumulation, indicated by extreme polygenic trait scores, in an additional 12.8% and 19.3% of overall cases of low- and high-HDL-C extremes, respectively. Thus, the genetic basis of extreme HDL-C concentrations encountered clinically is frequently polygenic, with contributions from both rare large-effect and common small-effect variants. Multiple types of genetic variants should be considered as contributing factors in patients with extreme dyslipidemia.
Asunto(s)
HDL-Colesterol/sangre , HDL-Colesterol/genética , Genotipo , Adulto , Anciano , Femenino , Variación Genética , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Masculino , Persona de Mediana EdadRESUMEN
OBJECTIVE: Next-generation sequencing technology is transforming our understanding of heterozygous familial hypercholesterolemia, including revision of prevalence estimates and attribution of polygenic effects. Here, we examined the contributions of monogenic and polygenic factors in patients with severe hypercholesterolemia referred to a specialty clinic. APPROACH AND RESULTS: We applied targeted next-generation sequencing with custom annotation, coupled with evaluation of large-scale copy number variation and polygenic scores for raised low-density lipoprotein cholesterol in a cohort of 313 individuals with severe hypercholesterolemia, defined as low-density lipoprotein cholesterol >5.0 mmol/L (>194 mg/dL). We found that (1) monogenic familial hypercholesterolemia-causing mutations detected by targeted next-generation sequencing were present in 47.3% of individuals; (2) the percentage of individuals with monogenic mutations increased to 53.7% when copy number variations were included; (3) the percentage further increased to 67.1% when individuals with extreme polygenic scores were included; and (4) the percentage of individuals with an identified genetic component increased from 57.0% to 92.0% as low-density lipoprotein cholesterol level increased from 5.0 to >8.0 mmol/L (194 to >310 mg/dL). CONCLUSIONS: In a clinically ascertained sample with severe hypercholesterolemia, we found that most patients had a discrete genetic basis detected using a comprehensive screening approach that includes targeted next-generation sequencing, an assay for copy number variations, and polygenic trait scores.
Asunto(s)
Hiperlipoproteinemia Tipo II/genética , Herencia Multifactorial , Mutación , Adulto , Anciano , Biomarcadores/sangre , LDL-Colesterol/sangre , Variaciones en el Número de Copia de ADN , Análisis Mutacional de ADN , Femenino , Dosificación de Gen , Marcadores Genéticos , Predisposición Genética a la Enfermedad , Pruebas Genéticas/métodos , Herencia , Heterocigoto , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Hiperlipoproteinemia Tipo II/sangre , Hiperlipoproteinemia Tipo II/diagnóstico , Masculino , Persona de Mediana Edad , Ontario , Fenotipo , Valor Predictivo de las Pruebas , Factores de Riesgo , Índice de Severidad de la EnfermedadRESUMEN
Significant resources have been invested in sequencing studies to investigate the role of rare variants in complex disease etiology. However, the diagnostic interpretation of individual rare variants remains a major challenge, and may require accurate variant functional classification and the collection of large numbers of variant carriers. Utilizing sequence data from 458 individuals with hypertriglyceridemia and 333 controls with normal plasma triglyceride levels, we investigated these issues using GCKR, encoding glucokinase regulatory protein. Eighteen rare non-synonymous GCKR variants identified in these 791 individuals were comprehensively characterized by a range of biochemical and cell biological assays, including a novel high-throughput-screening-based approach capable of measuring all variant proteins simultaneously. Functionally deleterious variants were collectively associated with hypertriglyceridemia, but a range of in silico prediction algorithms showed little consistency between algorithms and poor agreement with functional data. We extended our study by obtaining sequence data on family members; however, functional variants did not co-segregate with triglyceride levels. Therefore, despite evidence for their collective functional and clinical relevance, our results emphasize the low predictive value of rare GCKR variants in individuals and the complex heritability of lipid traits.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/genética , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Hiperlipoproteinemia Tipo IV/genética , Polimorfismo de Nucleótido Simple , Triglicéridos/sangre , Proteínas Adaptadoras Transductoras de Señales/química , Algoritmos , Animales , Células COS , Estudios de Casos y Controles , Chlorocebus aethiops , Variación Genética , Células HeLa , Humanos , Hiperlipoproteinemia Tipo IV/sangre , Ratones , Modelos Moleculares , Estructura Terciaria de Proteína , Análisis de Secuencia de ADNRESUMEN
OBJECTIVE: Estrogen deficiency is linked with increased low-density lipoprotein (LDL) cholesterol. The hormone receptor mediating this effect is unknown. G-protein estrogen receptor (GPER) is a recently recognized G-protein-coupled receptor that is activated by estrogens. We recently identified a common hypofunctional missense variant of GPER, namely P16L. However, the role of GPER in LDL metabolism is unknown. Therefore, we examined the association of the P16L genotype with plasma LDL cholesterol level. Furthermore, we studied the role of GPER in regulating expression of the LDL receptor and proprotein convertase subtilisin kexin type 9. APPROACH AND RESULTS: Our discovery cohort was a genetically isolated population of Northern European descent, and our validation cohort consisted of normal, healthy women aged 18 to 56 years from London, Ontario. In addition, we examined the effect of GPER on the regulation of proprotein convertase subtilisin kexin type 9 and LDL receptor expression by the treatment with the GPER agonist, G1. In the discovery cohort, GPER P16L genotype was associated with a significant increase in LDL cholesterol (mean±SEM): 3.18±0.05, 3.25±0.08, and 4.25±0.33 mmol/L, respectively, in subjects with CC (homozygous for P16), CT (heterozygotes), and TT (homozygous for L16) genotypes (P<0.05). In the validation cohort (n=339), the GPER P16L genotype was associated with a similar increase in LDL cholesterol: 2.17±0.05, 2.34±0.06, and 2.42±0.16 mmol/L, respectively, in subjects with CC, CT, and TT genotypes (P<0.05). In the human hepatic carcinoma cell line, the GPER agonist, G1, mediated a concentration-dependent increase in LDL receptor expression, blocked by either pretreatment with the GPER antagonist G15 or by shRNA-mediated GPER downregulation. G1 also mediated a GPER- and concentration-dependent decrease in proprotein convertase subtilisin kexin type 9 expression. CONCLUSIONS: GPER activation upregulates LDL receptor expression, probably at least, in part, via proprotein convertase subtilisin kexin type 9 downregulation. Furthermore, humans carrying the hypofunctional P16L genetic variant of GPER have increased plasma LDL cholesterol. In aggregate, these data suggest an important role of GPER in the regulation of LDL receptor expression and consequently LDL metabolism.
Asunto(s)
LDL-Colesterol/sangre , Mutación Missense , Receptores de Estrógenos/genética , Receptores de Estrógenos/metabolismo , Receptores Acoplados a Proteínas G/genética , Receptores Acoplados a Proteínas G/metabolismo , Adolescente , Adulto , Anciano , Canadá , Relación Dosis-Respuesta a Droga , Femenino , Regulación de la Expresión Génica , Frecuencia de los Genes , Genética de Población , Células Hep G2 , Heterocigoto , Homocigoto , Humanos , Masculino , Persona de Mediana Edad , Fenotipo , Proproteína Convertasa 9 , Proproteína Convertasas/genética , Proproteína Convertasas/metabolismo , Interferencia de ARN , Receptores de Estrógenos/efectos de los fármacos , Receptores Acoplados a Proteínas G/efectos de los fármacos , Receptores de LDL/genética , Receptores de LDL/metabolismo , Serina Endopeptidasas/genética , Serina Endopeptidasas/metabolismo , Transfección , Población Blanca/genética , Adulto JovenRESUMEN
PURPOSE OF REVIEW: To evaluate the potential clinical translation of high-throughput next-generation sequencing (NGS) methods in diagnosis and management of dyslipidemia. RECENT FINDINGS: Recent NGS experiments indicate that most causative genes for monogenic dyslipidemias are already known. Thus, monogenic dyslipidemias can now be diagnosed using targeted NGS. Targeting of dyslipidemia genes can be achieved by either: designing custom reagents for a dyslipidemia-specific NGS panel; or performing genome-wide NGS and focusing on genes of interest. Advantages of the former approach are lower cost and limited potential to detect incidental pathogenic variants unrelated to dyslipidemia. However, the latter approach is more flexible because masking criteria can be altered as knowledge advances, with no need for re-design of reagents or follow-up sequencing runs. Also, the cost of genome-wide analysis is decreasing and ethical concerns can likely be mitigated. DNA-based diagnosis is already part of the clinical diagnostic algorithms for familial hypercholesterolemia. Furthermore, DNA-based diagnosis is supplanting traditional biochemical methods to diagnose chylomicronemia caused by deficiency of lipoprotein lipase or its co-factors. SUMMARY: The increasing availability and decreasing cost of clinical NGS for dyslipidemia means that its potential benefits can now be evaluated on a larger scale.
Asunto(s)
Dislipidemias/genética , Secuenciación de Nucleótidos de Alto Rendimiento , Dislipidemias/diagnóstico , Exoma , Predisposición Genética a la Enfermedad , Estudio de Asociación del Genoma Completo , Humanos , Técnicas de Diagnóstico Molecular , Análisis de Secuencia de ADNRESUMEN
Joubert syndrome related disorders (JSRDs) have broad but variable phenotypic overlap with other ciliopathies. The molecular etiology of this overlap is unclear but probably arises from disrupting common functional module components within primary cilia. To identify additional module elements associated with JSRDs, we performed homozygosity mapping followed by next-generation sequencing (NGS) and uncovered mutations in TMEM237 (previously known as ALS2CR4). We show that loss of the mammalian TMEM237, which localizes to the ciliary transition zone (TZ), results in defective ciliogenesis and deregulation of Wnt signaling. Furthermore, disruption of Danio rerio (zebrafish) tmem237 expression produces gastrulation defects consistent with ciliary dysfunction, and Caenorhabditis elegans jbts-14 genetically interacts with nphp-4, encoding another TZ protein, to control basal body-TZ anchoring to the membrane and ciliogenesis. Both mammalian and C. elegans TMEM237/JBTS-14 require RPGRIP1L/MKS5 for proper TZ localization, and we demonstrate additional functional interactions between C. elegans JBTS-14 and MKS-2/TMEM216, MKSR-1/B9D1, and MKSR-2/B9D2. Collectively, our findings integrate TMEM237/JBTS-14 in a complex interaction network of TZ-associated proteins and reveal a growing contribution of a TZ functional module to the spectrum of ciliopathy phenotypes.
Asunto(s)
Enfermedades Cerebelosas/genética , Cilios/genética , Anomalías del Ojo/genética , Enfermedades Renales Quísticas/genética , Proteínas de la Membrana/genética , Mutación , Anomalías Múltiples , Adulto , Animales , Síndrome de Bardet-Biedl/genética , Caenorhabditis elegans/genética , Caenorhabditis elegans/ultraestructura , Estudios de Casos y Controles , Línea Celular , Cerebelo/anomalías , Niño , Preescolar , Mapeo Cromosómico , Cilios/metabolismo , Femenino , Expresión Génica , Técnicas de Silenciamiento del Gen , Técnicas de Inactivación de Genes , Estudios de Asociación Genética , Haplotipos , Humanos , Lactante , Recién Nacido , Masculino , Proteínas de la Membrana/metabolismo , Ratones , Microscopía Electrónica de Transmisión , Complejos Multiproteicos/metabolismo , Polimorfismo de Nucleótido Simple , Retina/anomalías , Análisis de Secuencia de ADN , Proteínas Wnt/metabolismo , Vía de Señalización Wnt , Pez Cebra/embriología , Pez Cebra/genéticaRESUMEN
AIMS: Activation of vascular GPER has been linked to vasodepressor effects in animals. However, the significance of GPER regulation on chronic blood pressure control in humans is unknown. METHODS: To examine this question we determined the functional significance of expression of a common missense single nucleotide variant of GPER, P16L in vascular smooth muscle cells, and its association with blood pressure in humans. Further, to validate the importance of carrying GPER P16L in the development of hypertension we assessed allele frequency in a cohort of hard-to-treat hypertensive patients referred to a tertiary care clinic. RESULTS: Expression of the GPER P16L variant (V) vs. wild type (WT) in rat aortic vascular smooth muscle cells, was associated with a significant decrease in G1 (1 µm, a GPER agonist)-mediated ERK phosphorylation (slope of the function of G1-stimulated ERK phosphorylation: GPER content WT: 16.2, 95% CI 9.9, 22.6; V: 5.0, 95% CI 1.0, 9.0; P < 0.005) and apoptosis (slope of the function of G1-stimulated apoptosis: GPER content: WT: 4.4, 95% CI: 3.4, 5.4; V: 2.5, 95% CI 1.6, 2.3 P < 0.005). Normotensive female subjects, but not male subjects, carrying this hypofunctional variant (allele frequency 22%) have increased blood pressure [mean arterial pressure: P16/P16: 80 ± 1 mmHg (n = 204) vs.â P16L carriers: 82 ± 1 mmHg (n = 127), 95% CI for difference: 0.6, 4.0 mmHg, P < 0.05], [systolic blood pressure: P16/P16: 105 ± 1 mmHg vs. P16L carriers: 108 ± 1 mmHg, 95% CI for difference:1.0, 5.1 mmHg, P < 0.05], [diastolic blood pressure: P16/P16: 66 ± 0.5 mmHg vs. P16L carriers 68 ± 0.7, 95% CI for difference: 0.2, 3.6 mmHg, P < 0.05]. Further, the P16L allele frequency was almost two-fold higher in female vs. male hypertensive patients (31% vs. 16%, allele ratio 0.5, 95% CI 0.32, 0.76, P < 0.05). CONCLUSIONS: The common genetic variant, GPER P16L, is hypofunctional and female carriers of this allele have increased blood pressure. There was an increased prevalence in a population of hard-to-treat hypertensive female patients. Cumulatively, these data suggest that in females, impaired GPER function might be associated with increased blood pressure and risk of hypertension.
Asunto(s)
Hipertensión/genética , Receptores de Estrógenos/genética , Receptores Acoplados a Proteínas G/genética , Adolescente , Adulto , Animales , Apoptosis , Células Cultivadas , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Femenino , Frecuencia de los Genes , Variación Genética , Humanos , Hipertensión/etiología , Masculino , Fosforilación , Ratas , Caracteres SexualesRESUMEN
OBJECTIVE: Earlier studies have suggested that a common genetic architecture underlies the clinically heterogeneous polygenic Fredrickson hyperlipoproteinemia (HLP) phenotypes defined by hypertriglyceridemia (HTG). Here, we comprehensively analyzed 504 HLP-HTG patients and 1213 normotriglyceridemic controls and confirmed that a spectrum of common and rare lipid-associated variants underlies this heterogeneity. METHODS AND RESULTS: First, we demonstrated that genetic determinants of plasma lipids and lipoproteins, including common variants associated with plasma triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) from the Global Lipids Genetics Consortium were associated with multiple HLP-HTG phenotypes. Second, we demonstrated that weighted risk scores composed of common TG-associated variants were distinctly increased across all HLP-HTG phenotypes compared with controls; weighted HDL-C and LDL-C risk scores were also increased, although to a less pronounced degree with some HLP-HTG phenotypes. Interestingly, decomposition of HDL-C and LDL-C risk scores revealed that pleiotropic variants (those jointly associated with TG) accounted for the greatest difference in HDL-C and LDL-C risk scores. The APOE E2/E2 genotype was significantly overrepresented in HLP type 3 versus other phenotypes. Finally, rare variants in 4 genes accumulated equally across HLP-HTG phenotypes. CONCLUSIONS: HTG susceptibility and phenotypic heterogeneity are both influenced by accumulation of common and rare TG-associated variants.
Asunto(s)
Hipertrigliceridemia/sangre , Hipertrigliceridemia/genética , Lípidos/sangre , Lípidos/genética , Adulto , Anciano , Alelos , Apolipoproteína E2/genética , Estudios de Casos y Controles , HDL-Colesterol/sangre , HDL-Colesterol/genética , LDL-Colesterol/sangre , LDL-Colesterol/genética , Femenino , Predisposición Genética a la Enfermedad , Variación Genética , Humanos , Hiperlipoproteinemia Tipo IV/sangre , Hiperlipoproteinemia Tipo IV/genética , Masculino , Persona de Mediana Edad , Herencia Multifactorial , Fenotipo , Factores de Riesgo , Triglicéridos/sangre , Triglicéridos/genéticaRESUMEN
Numerous single nucleotide polymorphisms (SNPs) have been found in recent genome wide association studies (GWAS) to be associated with subtle plasma triglyceride (TG) variation in normolipidemic subjects. However, since these GWAS did not specifically evaluate patients with rare disorders of lipoprotein metabolism--'hyperlipoproteinemia' (HLP)--it remains largely unresolved whether any of these SNP determinants of modest physiological changes in TG are necessarily also determinants of most HLP phenotypes. To address this question, we evaluated 28 TG-associated SNPs from GWAS in 386 unrelated adult patients with one of five Fredrickson phenotypes (HLP types 2A, 2B, 3, 4 and 5) and 242 matched normolipidemic controls. We found that several SNPs associated with TG in normolipidemic samples, including APOA5 p.S19W and -1131T>C, TRIB1 rs17321515, TBL2 rs17145738, GCKR rs780094, GALNT2 rs4846914 and ANGPTL3 rs12130333, were significantly associated with HLP types 2B, 3, 4 and 5. The findings indicate that: (i) the TG-associated Fredrickson HLP types 2B, 3, 4 and 5 are polygenic traits; (ii) these Fredrickson HLP types share numerous genetic determinants among themselves; and (iii) genetic determinants of modest TG variation in normolipidemic population samples also underlie--to an apparently even greater degree--susceptibility to these rare HLP phenotypes. Thus, the TG-associated Fredrickson HLP types 2B, 3, 4 and 5, although historically considered to be distinct are actually complex traits sharing among them several common genetic determinants seen in GWAS of normolipidemic population samples.
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Hiperlipoproteinemias/genética , Hipertrigliceridemia/genética , Herencia Multifactorial/genética , Proteínas Adaptadoras Transductoras de Señales/genética , Adulto , Anciano , Proteína 3 Similar a la Angiopoyetina , Proteínas Similares a la Angiopoyetina , Angiopoyetinas/genética , Apolipoproteína A-V , Apolipoproteínas A/genética , Proteínas Quinasas Dependientes de Calcio-Calmodulina , Femenino , Proteínas de Unión al GTP/genética , Frecuencia de los Genes , Estudio de Asociación del Genoma Completo , Genotipo , Humanos , Hiperlipoproteinemias/sangre , Hiperlipoproteinemias/patología , Hipertrigliceridemia/sangre , Hipertrigliceridemia/patología , Péptidos y Proteínas de Señalización Intracelular/genética , Desequilibrio de Ligamiento , Modelos Logísticos , Masculino , Persona de Mediana Edad , Análisis Multivariante , N-Acetilgalactosaminiltransferasas/genética , Fenotipo , Polimorfismo de Nucleótido Simple , Proteínas Serina-Treonina Quinasas/genética , Triglicéridos/sangre , Polipéptido N-AcetilgalactosaminiltransferasaRESUMEN
Apolipoprotein (apo) C-I is a constituent of chylomicrons, very low density lipoprotein, and high density lipoprotein. The role of apo C-I in human metabolism is incompletely defined. We took advantage of a naturally occurring amino acid polymorphism that is present in aboriginal North Americans, namely apo C-I T45S. We assessed the hypothesis that metabolic traits, including obesity-related and lipoprotein-related traits, would differ between carriers and noncarriers of apo C-I T45S. A genotyping assay was developed for APOC1 T45S and genotypes were determined in a sample of 410 Canadian Oji-Cree subjects. The allele frequency of the apo C-I S45 allele was approximately 8% in this sample. We observed the apo C-I S45 allele was significantly associated with 1) lower percent body fat (P < 0.05), 2) lower waist circumference (P = 0.058), 3) lower serum leptin levels (P < 0.05), and 4) lower plasma apo C-I levels (P < 0.0001), using a newly developed ELISA-based method. Taken together, these results suggest that at the whole human phenotype level, apo C-I is associated with the complex metabolic trait of obesity as well as with serum leptin levels.
Asunto(s)
Apolipoproteína C-I/sangre , Apolipoproteína C-I/genética , Indígenas Norteamericanos/genética , Leptina/sangre , Obesidad/genética , Polimorfismo Genético , Adolescente , Adulto , Composición Corporal , Canadá , Niño , Femenino , Frecuencia de los Genes , Estudios de Asociación Genética , Humanos , Hipertrigliceridemia/epidemiología , Masculino , Obesidad/sangre , Obesidad Abdominal/epidemiología , Prevalencia , Caracteres Sexuales , Circunferencia de la Cintura , Adulto JovenRESUMEN
Recent genome-wide association (GWA) studies have identified new genetic determinants of complex quantitative traits, including plasma triglyceride (TG). We hypothesized that common variants associated with mild TG variation identified in GWA studies would also be associated with severe hypertriglyceridemia (HTG). We studied 132 patients of European ancestry with severe HTG (fasting plasma TG > 10 mmol/l), who had no mutations found by resequencing of candidate genes, and 351 matched normolipidemic controls. We determined genotypes for: GALNT2 rs4846914, TBL2/MLXIPL rs17145738, TRIB1 rs17321515, ANGPTL3 rs12130333, GCKR rs780094, APOA5 rs3135506 (S19W), APOA5 rs662799 (-1131T > C), APOE (isoforms) and LPL rs328 (S447X). We found that: (i) genotypes, including those of APOA5 S19W, APOA5 -1131T > C, APOE, GCKR, TRIB1 and TBL2/MLXIPL, were significantly associated with severe HTG; (ii) odds ratios for these genetic variables were significant in both univariate and multivariate regression analyses, irrespective of the presence or absence of diabetes or obesity; (iii) a significant fraction-about one-quarter-of the explained variation in disease status was associated with these genotypes. Therefore, common SNPs (single nucleotide polymorphisms) that are associated with mild TG variation in GWA studies of normolipidemic subjects are also associated with severe HTG. Our findings are consistent with the emerging model of a complex genetic trait. At the extremes of a quantitative trait, such as severe HTG, are found the cumulative contributions of both multiple rare alleles with large genetic effects and common alleles with small effects.
Asunto(s)
Predisposición Genética a la Enfermedad , Hipertrigliceridemia/genética , Herencia Multifactorial , Proteínas Adaptadoras Transductoras de Señales/genética , Adulto , Apolipoproteínas/genética , Estudios de Casos y Controles , Femenino , Proteínas de Unión al GTP/genética , Genotipo , Humanos , Péptidos y Proteínas de Señalización Intracelular/genética , Masculino , Persona de Mediana Edad , Análisis Multivariante , N-Acetilgalactosaminiltransferasas/genética , Proteínas Serina-Treonina Quinasas/genética , Triglicéridos/sangre , Población Blanca/genética , Polipéptido N-AcetilgalactosaminiltransferasaRESUMEN
OBJECTIVES: To genotype common genetic variants found in the adipose triglyceride lipase (ATGL) gene and test them for association with cardiovascular disease risk factors in the Greenland Inuit. STUDY DESIGN: Candidate gene association study of discrete and quantitative traits related to cardiovascular health. METHODS: ATGL was sequenced in 10 European subjects to identify DNA sequence variants. The identified polymorphisms were subsequently genotyped in a population-based cohort of 1,218 unrelated Greenland Inuit subjects, ascertained from the Greenland Population Study. Genotypes and reconstructed haplotypes were tested for association with cardiovascular disease risk factors using additive, dominant or recessive models, corrected for age, sex and body mass index. RESULTS: Five single nucleotide polymorphisms and one 4-base pair deletion were identified in the European sample and were similarly polymorphic in the Greenland Inuit. Independently, variants were not associated with any cardiovascular traits. However, reconstructed rare ATGL haplotypes were associated with increased plasma triglyceride (TG) concentrations compared to the major haplotype under a dominant model (1.21+/-0.7 mmol/L and 1.11+/-0.6 mmol/L, respectively, p=0.006). CONCLUSIONS: Rare ATGL haplotypes are associated with increased plasma TG concentrations in the Greenland Inuit.
Asunto(s)
Haplotipos , Inuk/genética , Lipasa/genética , Polimorfismo Genético , Triglicéridos/sangre , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Enfermedades Cardiovasculares/genética , Femenino , Genotipo , Groenlandia , Humanos , Masculino , Persona de Mediana Edad , Factores de Riesgo , Adulto JovenRESUMEN
BACKGROUND: Familial hypercholesterolemia (FH) is a common autosomal codominant genetic disorder, which causes elevated levels of low-density lipoprotein cholesterol (LDL-C) and increased risk of premature atherosclerotic cardiovascular disease (ASCVD). Even among individuals with monogenic FH, there is substantial interindividual variability in LDL-C levels and risk of ASCVD. We assessed the influence of an LDL-C polygenic score on levels of LDL-C and risk of ASCVD for individuals with monogenic FH. METHODS: We constructed a weighted LDL-C polygenic score, composed of 28 single-nucleotide variants, for individuals with monogenic FH from the British Columbia FH (n=262); Nutrition, Metabolism and Atherosclerosis Clinic (n=552); and UK Biobank cohorts (n=306). We assessed the association between LDL-C polygenic score with LDL-C levels and ASCVD risk using linear regression and Cox-proportional hazard models, respectively. ASCVD was defined as myocardial infarction, coronary or carotid revascularization, transient ischemic attack, or stroke. The results from individual cohorts were combined in fixed-effect meta-analyses. RESULTS: Levels of LDL-C were significantly associated with LDL-C polygenic score in the Nutrition, Metabolism and Atherosclerosis Clinic cohort, UK Biobank cohort, and in the meta-analysis (ß [95% CI]=0.13 [0.072-0.19] per a 20% increase in LDL-C polygenic score percentile, P<0.0001). Additionally, an elevated LDL-C polygenic score (≥80th percentile) was associated with a trend towards increased ASCVD risk in all 3 cohorts individually. This association was statistically significant in the meta-analysis (hazard ratio [95% CI]=1.48 [1.02-2.14], P=0.04). CONCLUSIONS: Polygenic contributions to LDL-C explain some of the heterogeneity in clinical presentation and ASCVD risk for individuals with FH.
Asunto(s)
LDL-Colesterol/sangre , Hiperlipoproteinemia Tipo II/genética , Adulto , Factores de Riesgo de Enfermedad Cardiaca , Humanos , Hiperlipoproteinemia Tipo II/diagnóstico , Masculino , Persona de Mediana Edad , Herencia Multifactorial/genética , Polimorfismo de Nucleótido Simple , Modelos de Riesgos ProporcionalesRESUMEN
BACKGROUND: In 2013, our laboratory designed a targeted sequencing panel, "LipidSeq", to study the genetic determinants of dyslipidemia and metabolic disorders. Over the last 6 years, we have analyzed 3262 patient samples obtained from our own Lipid Genetics Clinic and international colleagues. Here, we highlight our findings and discuss research benefits and clinical implications of our panel. METHODS: LipidSeq targets 69 genes and 185 single-nucleotide polymorphisms (SNPs) either causally related or associated with dyslipidemia and metabolic disorders. This design allows us to simultaneously evaluate monogenic-caused by rare single-nucleotide variants (SNVs) or copy-number variants (CNVs)-and polygenic forms of dyslipidemia. Polygenic determinants were assessed using three polygenic scores, one each for low-density lipoprotein cholesterol, triglyceride, and high-density lipoprotein cholesterol. RESULTS: Among 3262 patient samples evaluated, the majority had hypertriglyceridemia (40.1%) and familial hypercholesterolemia (28.3%). Across all samples, we identified 24,931 unique SNVs, including 2205 rare variants predicted disruptive to protein function, and 77 unique CNVs. Considering our own 1466 clinic patients, LipidSeq results have helped in diagnosis and improving treatment options. CONCLUSIONS: Our LipidSeq design based on ontology of lipid disorders has enabled robust detection of variants underlying monogenic and polygenic dyslipidemias. In more than 50 publications related to LipidSeq, we have described novel variants, the polygenic nature of many dyslipidemias-some previously thought to be primarily monogenic-and have uncovered novel mechanisms of disease. We further demonstrate several tangible clinical benefits of its use.
Asunto(s)
Variaciones en el Número de Copia de ADN , Dislipidemias/genética , Herencia Multifactorial , Polimorfismo de Nucleótido Simple , Adulto , Anciano , HDL-Colesterol/sangre , HDL-Colesterol/genética , LDL-Colesterol/sangre , LDL-Colesterol/genética , Dislipidemias/sangre , Femenino , Humanos , Masculino , Persona de Mediana Edad , Triglicéridos/sangre , Triglicéridos/genéticaRESUMEN
BACKGROUND: Inhibitors of proprotein convertase subtilisin kexin 9 are indicated in Canada for treatment of patients with familial hypercholesterolemia (FH). Classically, FH is considered to be a monogenic condition caused by rare pathogenic mutations; however, some patients have hypercholesterolemia on a polygenic basis. Whether the effect of proprotein convertase subtilisin kexin 9 inhibitor treatment differs between patients with monogenic hypercholesterolemia and patients with polygenic hypercholesterolemia is unclear. METHODS: We performed retrospective chart reviews on patients treated with evolocumab 140 mg subcutaneously biweekly from the Lipid Genetics Clinic, London Health Sciences Centre. Evolocumab-treated patients with hypercholesterolemia were grouped into monogenic or polygenic categories on the basis of their genotype determined by targeted next-generation sequencing. Absolute and relative changes in low-density lipoprotein cholesterol (LDL-C) levels before and after evolocumab treatment were studied. RESULTS: In 32 patients with monogenic heterozygous FH and 7 patients with polygenic hypercholesterolemia treated with evolocumab, absolute incremental reductions in LDL-C were 2.94 ± 1.22 mmol/L and 3.15 ± 0.90 mmol/L, respectively (P = not significant), whereas percent reductions in LDL-C were 63.9% ± 16.0% and 67.7% ± 20.7%, respectively (P = not significant). CONCLUSION: Although the sample size is small, the findings suggest comparable biochemical responsiveness to evolocumab in both monogenic (heterozygous) and polygenic hypercholesterolemia.
INTRODUCTION: Au Canada, les inhibiteurs de la proprotéine convertase subtilisine/kexine de type 9 sont indiqués dans le traitement des patients atteints d'hypercholestérolémie familiale (HF). Traditionnellement, la HF est considérée comme une maladie monogénique causée par des mutations pathogènes rares. Toutefois, certains patients ont une hypercholestérolémie de forme polygénique. On ignore si les effets du traitement par inhibiteurs de la proprotéine convertase subtilisine/kexine de type 9 diffèrent entre les patients ayant une hypercholestérolémie monogénique et les patients ayant une hypercholestérolémie polygénique. MÉTHODES: Nous avons réalisé une revue rétrospective de dossiers de patients traités par évolocumab à raison de 140 mg par voie sous-cutanée 2 fois par semaine à la Lipid Genetics Clinic du London Health Sciences Centre. Les patients atteints d'hypercholestérolémie qui étaient traités par évolocumab ont été regroupés dans la catégorie de la forme monogénique et la catégorie de la forme polygénique en fonction de leur génotype déterminé par le séquençage ciblé de nouvelle génération. Nous avons étudié les changements absolus et relatifs des concentrations de cholestérol à lipoprotéines de faible densité (LDL-C) avant et après le traitement par évolocumab. RÉSULTATS: Chez 32 patients ayant une HF hétérozygote monogénique et 7 patients ayant une hypercholestérolémie polygénique traités par évolocumab, les réductions progressives absolues du LDL-C étaient respectivement de 2,94 ± 1,22 mmol/l et de 3,15 ± 0,90 mmol/l (P = non significatif), alors que les réductions du LDL-C en pourcentage étaient respectivement de 63,9 % ± 16,0 % et de 67,7 % ± 20,7 % (P = non significatif). CONCLUSION: Bien que la taille de l'échantillon soit petite, les résultats montrent une capacité de réponse biochimique comparable à l'évolocumab lors d'hypercholestérolémie (hétérozygote) monogénique et d'hypercholestérolémie polygénique.
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BACKGROUND: Coronary artery disease (CAD) risk traditionally has been assessed using clinical risk factors. We evaluated whether molecular genetic markers for CAD risk could add information to traditional variables. METHODS: We developed a false discovery rate 267-marker genetic risk score (FDR267) from markers that were significantly associated with CAD in the UK Biobank cohort meta-analysis. FDR267 was tested in the Atherosclerosis Risk in Communities cohort using logistic regression and Cox proportional hazards analyses in the European and African American groups. RESULTS: Our genetic risk score (FDR267) was associated with a 1.45 (95% confidence interval, 1.39-1.51) increase in odds ratio and a 1.32 (95% confidence interval, 1.26-1.38) increase in hazard ratio per standard deviation of the score. The score modestly improved the area under the curve (AUC) statistic when added to a clinical model (ΔAUC = 0.0112, P = 0.0002). FDR267 predicted incident CAD (C-index = 0.60), although it did not improve on clinical risk factors (ΔAUC = 0.0159, P = 0.0965). Individuals in the top quintile of FDR267 genetic risk were at approximately 2-fold increased risk compared with the bottom quintile, which is comparable to risk associated with self-reported family history. The performance of FDR267 was less robust in the African American sample. CONCLUSIONS: FDR267 is significantly associated with CAD in the European sample, with an effect size comparable to self-reported family history. FDR267 discriminated between individuals with and without CAD, but did not improve CAD risk prediction over clinical variables. FDR267 was less predictive of CAD risk in African Americans.
INTRODUCTION: L'évaluation du risque de maladie coronarienne (MC) a traditionnellement reposé sur les facteurs de risque cliniques. Nous avons évalué si les marqueurs génétiques moléculaires de risque de MC pourraient servir de complément aux variables traditionnelles. MÉTHODES: Nous avons élaboré un taux de fausses découvertes (FDR pour false discovery rate) du score de risque génétique du marqueur 267 (FDR267) provenant des marqueurs qui étaient associés de manière significative à la MC dans la méta-analyse de cohortes de la UK Biobank. Le FDR267 a été testé dans la cohorte du Atherosclerosis Risk in Communities à l'aide de la régression logistique et des analyses selon le modèle à risques proportionnels de Cox dans des groupes européens et afro-américains. RÉSULTATS: Notre score de risque génétique (FDR267) a été associé à une augmentation de 1,45 (intervalle de confiance [IC] à 95 %, 1,39-1,51) du rapport des cotes et d'une augmentation de 1,32 (IC à 95 %, 1,26-1,38) du risque relatif par l'écart-type des scores. Le score a modestement amélioré l'aire sous la courbe (ASC) lorsqu'il a été ajouté à un modèle clinique (ΔASC = 0,0112, P = 0,0002). Le FDR267 a prédit les nouveaux cas de MC (C-index [indice de concordance] = 0,60), mais il n'a pas amélioré les facteurs de risque cliniques (ΔASC = 0,0159, P = 0,0965). Les individus dans le quintile supérieur du risque génétique du FDR267 ont montré un risque accru d'environ 2 fois par rapport au quintile inférieur, soit un risque comparable au risque associé aux antécédents familiaux auto-rapportés. La performance du FDR267 s'est révélée moins robuste chez les Afro-Américains. CONCLUSIONS: Le FDR267 est associé de manière significative à la MC dans l'échantillon d'Européens et a une taille de l'effet comparable aux antécédents familiaux auto-rapportés. Le FDR267 a fait la discrimination entre les individus atteints ou non atteints de MC, mais n'a pas amélioré la prédiction du risque de MC par rapport aux variables cliniques. Le FDR267 a moins bien prédit le risque de MC chez les Afro-Américains.
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BACKGROUND: Familial hypercholesterolemia (FH) is an inherited condition of elevated serum low-density lipoprotein (LDL) cholesterol leading to premature coronary heart disease. We evaluated whether FH mutations are independently associated with the development of myocardial infarction (MI), after adjusting for LDL cholesterol level and clinical risk factors. METHODS: In 182 unrelated patients from different families referred with clinically suspected FH, targeted next-generation DNA sequencing was performed on 73 lipid-related genes and 178 single nucleotide polymorphisms, at 300-times mean read depth, to identify monogenic mutations and high-risk single nucleotide polymorphisms. RESULTS: Pathogenic FH mutations were identified in 27% of patients. Patients with mutations, compared with those without, were 12 years younger when referred to the lipid clinic (P < 0.001) and had higher baseline and post-treatment LDL cholesterol by 1.11 mmol/L (P < 0.001) and 0.62 mmol/L (P = 0.01), respectively. The hazard ratio for premature MI with respect to having an FH mutation, controlling for sex, hypertension, body mass index, diabetes, LDL cholesterol, and smoking, was 4.51 (P = 0.002). CONCLUSION: FH is a genetically diverse condition. FH mutations are independently associated with higher risk of premature MI in patients referred for hypercholesterolemia. Therefore, genotyping could guide cardiovascular risk stratification in the personalized treatment of FH.
INTRODUCTION: L'hypercholestérolémie familiale (HF) est une maladie héréditaire caractérisée par des concentrations élevées de lipoprotéines de faible densité (LDL) qui entraîne de manière précoce la maladie coronarienne. Nous avons évalué si les mutations propres à la HF sont indépendamment associées au développement de l'infarctus du myocarde (IM) après l'ajustement des concentrations de cholestérol LDL et des facteurs de risque cliniques. MÉTHODES: Chez les 182 patients non apparentés de différentes familles envoyés en consultation en raison d'une suspicion clinique de HF, nous avons réalisé le séquençage ciblé de l'ADN de nouvelle génération de 73 gènes du métabolisme des lipides et 178 polymorphismes mononucléotidiques, à une profondeur de lecture moyenne de 300 fois, pour déterminer les mutations monogènes et les polymorphismes mononucléotidiques à haut risque. RÉSULTATS: Nous avons déterminé les mutations pathogènes à l'origine de la HF de 27 % des patients. Comparativement aux patients qui ne présentaient pas de mutations, les patients qui en présentaient étaient 12 ans plus jeunes lorsqu'ils étaient envoyés en consultation à la clinique des lipides (P < 0,001) et avaient des concentrations respectives plus élevées de cholestérol LDL de 1,11 mmol/l (P < 0,001) et de 0,62 mmol/l (P = 0,01), au début et après le traitement. Le rapport de risque d'IM prématuré par rapport à la présence d'une mutation à l'origine de la HF, qui tient compte du sexe, de l'hypertension, de l'indice de masse corporelle, du diabète, du cholestérol LDL et de la consommation du tabac, était de 4,51 (P = 0,002). CONCLUSION: La HF est une maladie diversifiée sur le plan génétique. Les mutations à l'origine de la HF sont indépendamment associées à un risque plus élevé d'IM prématuré chez les patients envoyés en consultation en raison d'une hypercholestérolémie. Par conséquent, le génotypage pourrait être utile à la stratification du risque cardiovasculaire dans le traitement personnalisé de la HF.
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BACKGROUND: The rs9939609 T>A single-nucleotide polymorphism (SNP) in the FTO gene has previously been found to be associated with obesity in European Caucasian samples. The objective of this study is to examine whether this association extends to metabolic syndrome (MetS) and applies in non-Caucasian samples. METHODS: The FTO rs9939609 SNP was genotyped in 2121 subjects from four different non-Caucasian geographical ancestries. Subjects were classified for the presence or absence of MetS according to the International Diabetes Federation (IDF) and National Cholesterol Education Program Adult Treatment Panel (NCEP ATP) III definitions. RESULTS: Carriers of > or = 1 copy of the rs9939609 A allele were significantly more likely to have IDF-defined MetS (35.8%) than non-carriers (31.2%), corresponding to a carrier odds ratio (OR) of 1.23 (95% confidence interval [CI] 1.01 to 1.50), with a similar trend for the NCEP ATP III-defined MetS. Subgroup analysis showed that the association was particularly strong in men. The association was related to a higher proportion of rs9939609 A allele carriers meeting the waist circumference criterion; a higher proportion also met the HDL cholesterol criterion compared with wild-type homozygotes. CONCLUSION: Thus, the FTO rs9939609 SNP was associated with an increased risk for MetS in this multi-ethnic sample, confirming that the association extends to non-Caucasian population samples.
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Pueblo Asiatico/genética , Indígenas Norteamericanos/genética , Inuk/genética , Síndrome Metabólico/etnología , Síndrome Metabólico/genética , Polimorfismo de Nucleótido Simple , Proteínas/genética , Adulto , Dioxigenasa FTO Dependiente de Alfa-Cetoglutarato , Tamaño Corporal/genética , Canadá/epidemiología , HDL-Colesterol/sangre , Femenino , Frecuencia de los Genes , Predisposición Genética a la Enfermedad , Humanos , Masculino , Síndrome Metabólico/sangre , Persona de Mediana Edad , Oportunidad Relativa , Fenotipo , Medición de Riesgo , Factores de Riesgo , Factores SexualesRESUMEN
OBJECTIVE: The genetic determinants of severe hypertriglyceridemia (HTG; MIM 144650) in adults are poorly defined. We therefore resequenced 3 candidate genes, namely LPL, APOC2, and APOA5, to search for accumulation of missense mutations in patients with severe HTG compared with normolipidemic subjects. METHODS AND RESULTS: We resequenced >2 million base pairs of genomic DNA from 110 nondiabetic patients with severe HTG and determined the prevalence of coding sequence variants compared with 472 age- and sex-matched normolipidemic controls. We found: (1) heterozygous mutations (LPL p.Q-12E >11X, p.D25H, p.W86R, p.G188E, p.I194T and p.P207L; APOC2 p.K19T and IVS2-30G>A) in 10.0% of severe HTG patients compared with 0.2% of controls (carrier odds ratio [OR] 52, 95% confidence interval [CI] 8.6 to 319); and (2) an association of the APOA5 p.S19W missense variant with severe HTG (carrier OR 5.5 95% CI 3.3 to 9.1). Furthermore, either rare mutations or the APOA5 p.S19W variant were found in 41.8% of HTG subjects compared with 8.9% of controls (carrier OR 7.4, 95% CI 4.5 to 12.0). Also, heterozygotes for rare mutations had a significantly reduced plasma triglyceride response to fibrate monotherapy. CONCLUSIONS: Both common and rare DNA variants in candidate genes were found in a substantial proportion of severe HTG patients. The findings underscore the value of candidate gene resequencing to understand the genetic contribution in complex lipoprotein and metabolic disorders.