Genetic testing in specific cardiomyopathies

An increasing number of genetic tests for cardiomyopathies are becoming available for clinical use. This commentary will give a short overview of indications and challenges concerning genetic testing for these conditions

Degradation of the LDL receptors by PCSK9 is not mediated by a secreted protein acted upon by PCSK9 extracellularly

BACKGROUND: Proprotein convertase subtilisin/kexin type 9 (PCSK9) post-transcriptionally degrades the low density lipoprotein receptors (LDLR). However, it is unknown whether PCSK9 acts directly on the LDLR or if PCSK9 activates another protein that in turn causes degradation of the LDLR. RESULTS: We have transiently transfected HepG2 cells with wild-type and mutant D374Y-PCSK9 plasmids to study the effect of the conditioned medium on the LDLR of untransfected HepG2 cells. The ability of the conditioned medium to reduce the internalization of LDL was abolished by removal of recombinant PCSK9 from the conditioned medium by affinity chromatography. Thus, PCSK9 is the only factor in the conditioned medium able to mediate degradation of the LDLR. Moreover, fractionation of the conditioned medium by gel filtration showed that the ability of the fractions to reduce the internalization of LDL, closely paralleled the amount of D374Y-PCSK9 in the fractions. Incubation of a secreted, truncated LDLR without cytoplasmic and transmembrane domains, as well as membrane fractions from HepG2 cells, with conditioned medium containing PCSK9, did not reduce the amount of LDLR as determined by western blot analysis. Thus, the LDLR is not degraded by PCSK9 on the cell surface. The LDLR of HepG2 cells incubated with conditioned medium was protected from PCSK9-mediated degradation by the addition of nocodazole or ammonium chloride, but was not protected when the conditioned medium was made hypertonic. These findings indicate that the intracellular degradation of the LDLR involves intracellular transport along microtubules, an acidic intracellular compartment and that it occurs even when endocytosis through clathrin-coated pits has been blocked. CONCLUSION: Degradation of the LDLR by PCSK9 is not mediated by a secreted protein acted upon by PCSK9 extracellularly. Also the PCSK9-mediated degradation of the LDLR does not take place on the cell surface. Rather, the PCSK9-mediated degradation of the LDLR appears to take place intracellularly and occurs even when endocytosis through clathrin-coated pits is blocked by hypertonic medium

Left ventricular mechanical dispersion by tissue Doppler imaging: a novel approach for identifying high-risk individuals with long QT syndrome

Forutsigelse av livstruende hjerterytmeforstyrrelser Hjertespesialist og forsker Kristina Hermann Haugaa har i sin doktorgrad funnet en ny metode som kan brukes til å forutsi hvilke pasienter som kommer til å få alvorlige hjerterytmeforstyrrelser: Ultralyd av hjertet med ny metode kan avsløre hvem som har risiko for hjerterytmeforstyrrelser og hvem som ikke har det. Plutselig hjertedød på grunn av hjerterytmeforstyrrelser er en av de vanligste dødsårsakene i Norge og i den øvrige vestlige verden. Den største risikogruppen er personer som har hatt hjerteinfarkt. Plutselig hjertedød hos yngre skyldes ofte arvelige hjertesykdommer. I avhandlingen “Prediction of cardiac ventricular arrhythmias by echocardiography in patients at risk” undersøker Kristina Haugaa både yngre pasienter med arvelige hjerterytmeforstyrrelser og pasienter som har gjennomgått hjerteinfarkt med den nye metoden for hjerteultralyd. Pasientene ble fulgt i over to år etter hjerteinfarkt. Studiene viser at ujevn hjertekontraksjon er en risikomarkør for å få hjerterytmeforstyrrelser og at den nye metoden vurderer risikoen bedre enn dagens metoder. Med bedre risikovurdering kan man bedre fordele resursene for behandling. Behandlingen innebærer oftest at pasientene i tillegg til medisin får operert inn en automatisk hjertestarter. Den nye metoden som er brukt i avhandlingen vil kunne forbedre utvelgelsen av pasienter med høy risiko for død slik at disse kan utstyres med hjertestarter

Subjects with Molecularly Defined Familial Hypercholesterolemia or Familial Defective apoB-100 Are Not Being Adequately Treated

To study whether subjects with a molecular genetic diagnosis of familial hypercholesterolemia (FH) or familial defective apoB-100 (FDB) are being adequately treated.A questionnaire regarding medical history was sent to 2611 subjects who had been provided with a molecular genetic diagnosis of FH or FDB, and a blood sample was obtained for lipid measurements.956 (36.6%) of the 2611 subjects participated. The mean age for starting lipid-lowering therapy was 33.4 (±12.1) years. Among those below 18 years of age, only 20.4% were on lipid-lowering drugs, whereas 89.1% of those aged 18 and above were on lipid-lowering drugs. The mean levels of total serum cholesterol and LDL-cholesterol were 5.7 (±1.5) mmol/l and 3.9 (±1.3) mmol/l, respectively. Among those who were on lipid-lowering drugs, 29.0% and 12.2% had levels of LDL cholesterol below 3.0 mmol/l and 2.6 mmol/l, respectively. Only 47.3% of the 956 subjects were considered as being adequately treated largely due to a failure to titrate their drug regimens. From the use of cholesterol-years score, lipid-lowering therapy must start before the age of 20 in order to prevent the subjects from contracting premature coronary heart disease.The majority of FH/FDB subjects are being diagnosed late in life and are not being adequately treated. In order to prevent them from contracting premature coronary heart disease, it is key that levels of LDL cholesterol are normalized from a young age and that sufficient doses of lipid-lowering drugs are being used

Molecular analysis of three known and one novel LPL variants in patients with type I hyperlipoproteinemia.

Abstract Background and aims Type I hyperlipoproteinemia, also known as familial chylomicronemia syndrome (FCS), is a rare autosomal recessive disorder caused by variants in LPL, APOC2, APOA5, LMF1 or GPIHBP1 genes. The aim of this study was to identify novel variants in the LPL gene causing lipoprotein lipase deficiency and to understand the molecular mechanisms. Methods and results A total of 3 individuals with severe hypertriglyceridemia and recurrent pancreatitis were selected from the Lipid Clinic at Sahlgrenska University Hospital and LPL was sequenced. In vitro experiments were performed in human embryonic kidney 293T/17 (HEK293T/17) cells transiently transfected with wild type or mutant LPL plasmids. Cell lysates and media were used to analyze LPL synthesis and secretion. Media were used to measure LPL activity. Patient 1 was compound heterozygous for three known variants: c.337T > C (W113R), c.644G > A (G215E) and c.1211T > G (M404R); patient 2 was heterozygous for the known variant c.658A > C (S220R) while patient 3 was homozygous for a novel variant in the exon 5 c.679G > T (V227F). All the LPL variants identified were loss-of-function variants and resulted in a substantial reduction in the secretion of LPL protein. Conclusion We characterized at the molecular level three known and one novel LPL variants causing type I hyperlipoproteinemia showing that all these variants are pathogenic

A Novel SCN5A Mutation in a Patient with Coexistence of Brugada Syndrome Traits and Ischaemic Heart Disease

Brugada syndrome (BrS) is a primary electrical heart disease, which can lead to sudden cardiac death. In older patients with BrS, the disease may coexist with ischaemic heart disease (IHD) and recent studies support a synergistic proarrhythmic effect of the two disease entities. We report a case that illustrates this. The index patient was a middle-aged patient with BrS traits, IHD, and aborted sudden cardiac death. Mutation analysis discovered a novel mutation P468L in the NaV1.5 sodium channel. Surprisingly, voltage-clamp experiments on the wild-type and mutant NaV1.5 channels expressed in HEK cells revealed no functional effect of the mutation. In a patient like ours, the distinction between IHD and BrS as the cause of an aborted sudden cardiac death is hard to establish and mounting evidence shows that coexistence of the two may have a synergistic proarrhythmic effect

Identification and characterization of two novel mutations in the LPL gene causing type I hyperlipoproteinemia

Background Type 1 hyperlipoproteinemia is a rare autosomal recessive disorder most often caused by mutations in the lipoprotein lipase (LPL) gene resulting in severe hypertriglyceridemia and pancreatitis. Objectives The aim of this study was to identify novel mutations in the LPL gene causing type 1 hyperlipoproteinemia and to understand the molecular mechanisms underlying the severe hypertriglyceridemia. Methods Three patients presenting classical features of type 1 hyperlipoproteinemia were recruited for DNA sequencing of the LPL gene. Pre-heparin and post-heparin plasma of patients were used for protein detection analysis and functional test. Furthermore, in\ua0vitro experiments were performed in HEK293\ua0cells. Protein synthesis and secretion were analyzed in lysate and medium fraction, respectively, whereas medium fraction was used for functional assay. Results We identified two novel mutations in the LPL gene causing type 1 hyperlipoproteinemia: a two base pair deletion (c.765_766delAG) resulting in a frameshift at position 256 of the protein (p.G256TfsX26) and a nucleotide substitution (c.1211\ua0T\ua0>\ua0G) resulting in a methionine to arginine substitution (p.M404\ua0R). LPL protein and activity were not detected in pre-heparin or post-heparin plasma of the patient with p.G256TfsX26 mutation or in the medium of HEK293\ua0cells over-expressing recombinant p.G256TfsX26 LPL. A relatively small amount of LPL p.M404\ua0R was detected in both pre-heparin and post-heparin plasma and in the medium of the cells, whereas no LPL activity was detected. Conclusions We conclude that these two novel mutations cause type 1 hyperlipoproteinemia by inducing a loss or reduction in LPL secretion accompanied by a loss of LPL enzymatic activity

Serum lipids, apoproteins and nutrient intake in rural Cretan boys consuming high-olive-oil diets

A high intake of olive oil has produced high levels of high-density and low levels of low-density lipoprotein cholesterol in short-term dietary trials. To investigate long-term effects of olive oil we have studied the diet and serum lipids of boys in Crete, where a high olive oil consumption is the norm. Seventy-six healthy rural Cretan boys aged 7–9 years were studied. The diet was assessed by a 2-day dietary recall. Blood was collected according to a standardized protocol and sera were analyzed in a rigidly standardized laboratory. The mean daily intake of energy was 11.0 MJ (2629 kcal). The intake of fat (45.0% of energy) and oleic acid (27.2% of energy) was high, and that of saturated fat low (10.0% of energy), reflecting a high consumption of olive oil. The high consumption of olive oil was confirmed by a high proportion of oleic-acid (27.1 %) in serum cholesteryl fatty acids. Mean concentration of serum total cholesterol was 4.42 mmol 1−1 (171 mg dl−1 ), of HDL-cholesterol 1.40 mmol 1−1 (54 mg dl−1), of serum triglycerides 0.59 mmol I−1 (52 mg dl−1 ), of apo-A1 1210 mg 1−1 and of LDL apo-B 798 mg 1−1. The body mass index of the Cretan boys (18.2 kg m−2) was on average 2 kg m−2 higher than that of boys from other countries. Contrary to our expectation, the Cretan boys did not show a more favourable serum lipoprotein pattern than boys from more westernized countries studied previously using the same protocol. Our hypothesis that a typical, olive-oil-rich Cretan diet causes a relatively high HDL- to total cholesterol ratio is not supported by the present findings

Leisure time physical activity in middle age predicts the metabolic syndrome in old age: results of a 28-year follow-up of men in the Oslo study

<p>Abstract</p> <p>Background</p> <p>Data are scarce on the long term relationship between leisure time physical activity, smoking and development of metabolic syndrome and diabetes. We wanted to investigate the relationship between leisure time physical activity and smoking measured in middle age and the occurrence of the metabolic syndrome and diabetes in men that participated in two cardiovascular screenings of the Oslo Study 28 years apart.</p> <p>Methods</p> <p>Men residing in Oslo and born in 1923–32 (n = 16 209) were screened for cardiovascular diseases and risk factors in 1972/3. Of the original cohort, those who also lived in same area in 2000 were invited to a repeat screening examination, attended by 6 410 men. The metabolic syndrome was defined according to a modification of the National Cholesterol Education Program criteria. Leisure time physical activity, smoking, educational attendance and the presence of diabetes were self-reported.</p> <p>Results</p> <p>Leisure time physical activity decreased between the first and second screening and tracked only moderately between the two time points (Spearman's ρ = 0.25). Leisure time physical activity adjusted for age and educational attendance was a significant predictor of both the metabolic syndrome and diabetes in 2000 (odds ratio for moderately vigorous versus sedentary/light activity was 0.65 [95% CI, 0.54–0.80] for the metabolic syndrome and 0.68 [0.52–0.91] for diabetes) (test for trend P < 0.05). However, when adjusted for more factors measured in 1972/3 including glucose, triglycerides, body mass index, treated hypertension and systolic blood pressure these associations were markedly attenuated. Smoking was associated with the metabolic syndrome but not with diabetes in 2000.</p> <p>Conclusion</p> <p>Physical activity during leisure recorded in middle age prior to the current waves of obesity and diabetes had an independent predictive association with the presence of the metabolic syndrome but not significantly so with diabetes 28 years later in life, when the subjects were elderly.</p