Giovanni Battista Vigna

Efficacy and safety of a microsomal triglyceride transfer protein inhibitor in patients with homozygous familial hypercholesterolaemia: a single-arm, open-label, phase 3 study

BACKGROUND Patients with homozygous familial hypercholesterolaemia respond inadequately to existing drugs. We aimed to assess the efficacy and safety of the microsomal triglyceride transfer protein inhibitor lomitapide in adults with this disease. METHODS We did a single-arm, open-label, phase 3 study of lomitapide for treatment of patients with homozygous familial hypercholesterolemia. Current lipid lowering therapy was maintained from 6 weeks before baseline through to at least week 26. Lomitapide dose was escalated on the basis of safety and tolerability from 5 mg to a maximum of 60 mg a day. The primary endpoint was mean percent change in levels of LDL cholesterol from baseline to week 26, after which patients remained on lomitapide through to week 78 for safety assessment. Percent change from baseline to week 26 was assessed with a mixed linear model. FINDINGS 29 men and women with homozygous familial hypercholesterolaemia, aged 18 years or older, were recruited from 11 centres in four countries (USA, Canada, South Africa, and Italy). 23 of 29 enrolled patients completed both the efficacy phase (26 weeks) and the full study (78 weeks). The median dose of lomitapide was 40 mg a day. LDL cholesterol was reduced by 50% (95% CI -62 to -39) from baseline (mean 8·7 mmol/L [SD 2·9]) to week 26 (4·3 mmol/L [2·5]; p<0·0001). Levels of LDL cholesterol were lower than 2·6 mmol/L in eight patients at 26 weeks. Concentrations of LDL cholesterol remained reduced by 44% (95% CI -57 to -31; p<0·0001) at week 56 and 38% (-52 to -24; p<0·0001) at week 78. Gastrointestinal symptoms were the most common adverse event. Four patients had aminotransaminase levels of more than five times the upper limit of normal, which resolved after dose reduction or temporary interruption of lomitapide. No patient permanently discontinued treatment because of liver abnormalities. INTERPRETATION Our study suggests that treatment with lomitapide could be a valuable drug in the management of homozygous familial hypercholesterolaemia. FUNDING FDA Office of the Orphan Product Development, Aegerion Pharmaceuticals.

Role of Anti-Oxidants in Atherosclerosis: Epidemiological and Clinical Update

Low density lipoprotein (LDL) oxidative modification in the vascular wall seems to be a key factor in atherosclerosis development. Oxidised LDLs might recruit monocytes and favour their transformation into foam cells through a receptor-mediated intake (scavenger pathway). Moreover oxidised LDLs show cytotoxic potential which is probably responsible for endothelial cell damage and macrophage degeneration in the atherosclerotic human plaque. Following the oxidation hypothesis of atherosclerosis the role of natural antioxidants, i.e. Vitamin C, Vitamin E and carotenoids, has been investigated in a large number of epidemiological, clinical and experimental studies. Animal studies indicate that dietary antioxidants may reduce atherosclerosis progression, and observational data in humans suggest that antioxidant vitamin ingestion is associated with reduced cardiovascular disease, but the results of randomised controlled trials are mainly disappointing. It has been suggested that natural antioxidants may be effective only in selected subgroups of patients with high levels of oxidative stress or depletion of natural antioxidant defence systems. The favourable effects shown by some studies relating antioxidant dietary intake and cardiovascular disease, may have been exerted by other chemicals present in foods. Flavonoids are the ideal candidates, since they are plentiful in foods containing antioxidant vitamins (i.e. fruits and vegetables) and are potent antioxidants. Tea and wine, rich in flavonoids, seem to have beneficial effects on multiple mechanisms involved in atherosclerosis. Future studies should probably select patients in a context of high-oxidative stress / low-antioxidant defence, to verify if antioxidants may really prove useful as therapeutic anti-atherosclerotic agents.

Effect of a Standardized Grape Seed Extract on Low-Density Lipoprotein Susceptibility to Oxidation in Heavy Smokers

The aim of our study was to evaluate the effect of a standardized formulation of a polyphenolic extract of grapes (Leucoselect-Phytosome [LP]) on low-density lipoprotein (LDL) susceptibility to oxidation in a group of heavy smokers. A randomized, double-blind, crossover study was undertaken in 24 healthy male heavy smokers, aged > or = 50 years. Enrolled subjects were given 2 capsules twice daily for 4 weeks (phase 1). Each capsule contained 75 mg of a grape procyanidin extracts and soy-phosphatidlcholine or placebo consisiting of 75 mg lactose and soy-phosphatidlcholine. A wash out period of 3 weeks was then followed by 4 weeks of the opposite treatment (phase 2). Blood samples were taken at baseline and at the end of each phase and assayed for plasma lipids and LDL susceptibility to oxidation. Compliance was good, and no adverse effects were recorded. Subjects did not show significant modification of total cholesterol (TC), triglycerides (TG), high-density lipoprotein-cholesterol (HDL-C) and LDL-C during LP treatment. Among oxidative indices, thiobarbituric acid reactive substances (TBARS) concentration was significantly reduced in subjects taking LP (-14.7% +/- 21.1% v +5.0% +/- 18.1%, P <.01), and the lag phase prolonged (+15.4% +/- 24.4% v -0.1% +/- 16.0%, P <.05) compared with placebo and basal values. The antioxidant potential of grape seed extract polyphenols may prove effective in a model of oxidative stress (smoking); however more investigational data are needed before use in wider clinical settings.

Prevalence of ANGPTL3 and APOB Gene Mutations in Subjects With Combined Hypolipidemia

Objective—Mutations of the ANGPTL3 gene have been associated with a novel form of primary hypobetalipoproteinemia, the combined hypolipidemia (cHLP), characterized by low total cholesterol and low HDL-cholesterol levels. The aim of this work is to define the role of ANGPTL3 gene as determinant of the combined hypolipidemia phenotype in 2 large cohorts of 913 among American and Italian subjects with primary hypobetalipoproteinemia (total cholesterol <5th percentile). Methods and Results—The combined hypolipidemia cut-offs were chosen according to total cholesterol and HDL-cholesterol levels reported in the ANGPTL3 kindred described to date: total cholesterol levels, <2nd percentile and HDL-cholesterol, levels <2nd decile. Seventy-eight subjects with combined hypolipidemia were analyzed for ANGPTL3 and APOB genes. We identified nonsense and/or missense mutations in ANGPTL3 gene in 8 subjects; no mutations of the APOB gene were found. Mutated ANGPTL3 homozygous/compound heterozygous subjects showed a more severe biochemical phenotype compared to heterozygous or ANGPTL3 negative subjects, although ANGPTL3 heterozygotes did not differ from ANGPTL3 negative subjects. Conclusion—These results demonstrated that in a cohort of subjects with severe primary hypobetalipoproteinemia the prevalence of ANGPTL3 gene mutations responsible for a combined hypolipidemia phenotype is about 10%, whereas mutations of APOB gene are absent.

Lipids and other risk factors selected by discriminant analysis in symptomatic patients with supra-aortic and peripheral atherosclerosis

BackgroundDifferent patterns of risk factors might be related to the involvement of specific vascular districts by atherosclerosis. In this sense, many investigations have addressed coronary artery disease, whereas extracoronary atherosclerosis has received less extensive attention. Methods and ResultsVascular risk factors, with particular attention to lipid parameters (total cholesterol [TC]; triglycerides; high density lipoprotein cholesterol [HDL-C], HDI2 and HDL3 cholesterol [HDL2-C, HDL3-C]), were evaluated by means of univariate and multivariate (discriminant) analysis in a group of 169 patients (128 men and 41 women; mean ages, 58±7 and 62±7 years, respectively) with clinically and angiographically demonstrated atherosclerosis of the supra-aortic trunk and/or lower limbs. Patients with coronary artery disease were excluded from this study. The control group consisted of 140 age- and sex-matched individuals. By univariate analysis, smoking was more closely associated with peripheral atherosclerosis, whereas blood pressure was higher in patients with supra-aortic disease. Unrecognized diabetes mellitus was a frequent finding in patients with peripheral disease. The percentage of hyperlipidemias was fourfold higher in patients than in control subjects, with differences consisting of higher triglycerides and lower HDL-C, HDL2-C, and HDL3-C concentrations. By discriminant analysis, high correct classification (CC) rates were achieved in the various patient subgroups on the basis of variables selected from the statistical function. In male patients with peripheral disease, the variables HDL-C, smoking, diastolic blood pressure, uric acid, and glucose, in that order, yielded a CC in 90.4% of the cases; in female patients, smoking, TC/HDL-C, and body mass index gave a CC rate of 95.9%. In men with cerebral disease, the selected variables TC/HDL-C, diastolic blood pressure, and TC yielded a CC of 90.7%; in women, uric acid, TC/HDL-C, and fibrinogen levels produced a CC rate of 89.2%. ConclusionsRisk profiles in atherosclerosis of the supra-aortic trunks and lower limbs seem to differ in relation to gender and circulatory district involved. The importance of lipid parameters, in particular HDL-C, HDL2-C, and TC/HDL-C, as extracoronary risk factors is further confirmed.

Lipoprotein(a), Inflammation, and Peripheral Arterial Disease in a Community-Based Sample of Older Men and Women (the InCHIANTI Study)

Lipoprotein(a) (Lp[a]) may represent an independent risk factor for peripheral arterial disease of the lower limbs (LL-PAD), but prospective data are scant. We estimated the association between baseline Lp(a) with prevalent and incident LL-PAD in older subjects from the InCHIANTI Study. LL-PAD, defined as an ankle-brachial index <0.90, was assessed at baseline and over a 6-year follow-up in a sample of 1,002 Italian subjects 60 to 96 years of age. Plasma Lp(a) and potential traditional and novel cardiovascular risk factors (including a score based on relevant inflammatory markers) were entered in multivariable models to assess their association with prevalent and incident LL-PAD. At baseline, Lp(a) concentration was directly related to the number of increased inflammatory markers (p <0.05). There were 125 (12.5%) prevalent cases of LL-PAD and 57 (8.3%) incident cases. After adjustment for potential confounders, participants in the highest quartile of the Lp(a) distribution (>/=32.9 mg/dl) were more likely to have LL-PAD compared to those in the lowest quartile (odds ratio [OR] 1.83, 95% confidence interval [CI] 1.01 to 3.33). The association was stronger (OR 3.80, 95% CI 1.50 to 9.61) if LL-PAD was defined by harder criteria, namely an ankle-brachial index <0.70. Compared to subjects in the lowest Lp(a) quartile, those in the highest quartile showed a somewhat increased risk of incident LL-PAD (lowest quartile 7.7%, highest quartile 10.8%), but the association was not statistically significant (OR 1.52, 95% CI 0.71 to 3.22). In conclusion, Lp(a) is an independent LL-PAD correlate in the cross-sectional evaluation, but further prospective studies in larger populations, with longer follow-up and more definite LL-PAD ranking, might be needed to establish a longitudinal association.

Risk of hospitalization for upper gastrointestinal tract bleeding

OBJECTIVE This study evaluates the hospitalization risk for upper gastrointestinal bleeding (UGIB) with reference to the clinical characteristics of patients and drugs taken before admission. METHODS This study is based on the GIFA (Italian Group for the Pharmacosurveillance in the Elderly) database. Cases with an ICD-9 code of esophagus, stomach or duodenum bleeding, or acute esophago-gastroduodenal disease associated with anemia have been classified as UGIB. Sex, age, year of observation, drugs taken at home, comorbidity, smoking, alcohol, and use of gastroprotectants have been also taken into account. Statistical analysis has been conducted using multivariate logistic regression models. RESULTS 32,388 patients have been enrolled, 940 of which presented UGIB. Age, comorbidity, use of smoke and alcohol, hospitalization duration, and mortality during hospitalization were significantly higher in UGIB than nonUGIB patients. Increased UGIB risk has been found in patients taking NSAIDs (both when aspirin was included or excluded), acetaminophen, constipating agents, iron, ethacrynic acid, propranolol. Reduced UGIB risk has been found in patients taking nitrates. CONCLUSIONS UGIB risk appears to correlate with clinical characteristics of the patient: it increases with age, comorbidity, and smoke and alcohol consumption. Among drugs, NSAIDs are associated with the highest UGIB risk, while nitrates with a reduction of risk.

Novel mutations of CETP gene in Italian subjects with hyeralphalipoproteinemia

Cholesteryl ester transfer protein (CETP) is a plasma glycoprotein that catalyses the transfer of cholesteryl esters from HDL to the other plasma lipoproteins. Genetic deficiency of CETP is one of the known causes of elevation of plasma HDL-C (primary hyperalphalipoproteinemia, HALP). We sequenced CETP gene in a group of 24 Italian subjects with primary HALP (HDL-C>80 mg/dl) suspected to have CETP deficiency. Two unrelated subjects both coming from the same geographical district, were found to be heterozygous for a nucleotide substitution in exon 6 (c.544C>T) and another subject was found to be heterozygous for a C>T transition in exon 9 (c.802C>T). Both mutations introduce a premature stop codon and are predicted to cause the production of truncated proteins (Q165X and R268X, respectively) devoid of function. The fourth proband was found to carry a T>C substitution in intron 15 (c.1407+2T>C) predicted to abolish the function of the donor splice site. To define the effect of this mutation on CETP pre-mRNA splicing we analysed CETP mRNA in COS-1 cells expressing a CETP minigene harbouring the mutation. The analysis of minigene transcript in COS-1 cells showed that IVS15+2T>C mutation caused the formation of an abnormal mRNA in which exon 14 joins directly to exon 16, predicted to encode a truncated peptide of 435 amino acids. In mutation carriers plasma CETP activity was found to be reduced by 38-60%. These are the first mutations in the CETP gene found in Italian subjects with HALP.

Targeted lipidomics distinguishes patient subgroups in mild cognitive impairment (MCI) and late onset Alzheimer's disease (LOAD).

Background Diverse research approaches support the concept that a clinical diagnosis of Late-Onset Alzheimers Disease (LOAD) does not distinguish between subpopulations with differing neuropathologies, including dementia patients with amyloid deposition and dementia patients without amyloid deposition but with cortical thinning. Mild cognitive impairment (MCI) is generally considered the prodromal phase for LOAD, however, while a number of studies have attempted to define plasma biomarkers for the conversion of MCI to LOAD, these studies have not taken into account the heterogeneity of patient cohorts within a clinical phenotype. Methods Studies of MCI and LOAD in several laboratories have demonstrated decrements in ethanolamine plasmalogen levels in plasma and brain and increased levels of diacylglycerols in plasma and brain. To further extend these studies and to address the issue of heterogeneity in MCI and LOAD patient groups we investigated the levels of diacylglycerols and ethanolamine plasmalogens in larger cohorts of patients utilizing, high-resolution (0.2 to 2 ppm mass error) mass spectrometry. Results For the first time, our lipidomics data clearly stratify both MCI and LOAD subjects into 3 different patient cohorts within each clinical diagnosis. These include i) patients with lower circulating ethanolamine plasmalogen levels; ii) patients with augmented plasma diacylglycerol levels; and iii) patients with neither of these lipid alterations. Conclusions These represent the first serum biochemical data to stratify MCI and LOAD patients, advancing efforts to biochemically define patient heterogeneity in cognitive disorders. General significance Lipidomics offers a new approach for identifying biomarkers and biological targets in cognitive disorders.

Long-Term Efficacy and Safety of the Microsomal Triglyceride Transfer Protein Inhibitor Lomitapide in Patients With Homozygous Familial Hypercholesterolemia

Homozygous familial hypercholesterolemia is a genetic disorder characterized by low-density lipoprotein (LDL)-receptor dysfunction, markedly elevated levels of LDL-cholesterol (LDL-C) and premature atherosclerosis. Patients are often poorly responsive to conventional lipid-lowering therapies that upregulate LDL-receptor expression.1 Lomitapide inhibits microsomal triglyceride transfer protein, which lipidates nascent apolipoprotein (apo)B-containing lipoproteins. In a pivotal 78-week open-label trial, lomitapide, titrated to the maximal tolerable dose, decreased LDL-C by 50% at the end of the efficacy phase (week 26) in patients with homozygous familial hypercholesterolemia.2 The principal adverse events included gastrointestinal disturbances, hepatic enzyme elevations, and increased liver fat. Here we provide additional long-term efficacy and safety data, including an exploratory analysis of the potential metabolic consequences of hepatic fat accumulation from an extension trial (NCT00943306). Patients continued on lomitapide at the maximally tolerated dose until transition to commercial or compassionate lomitapide. Lipid-lowering therapies, including apheresis, could be modified at the investigator’s discretion if LDL-C was <100 mg/dL. Both studies received institutional review board and regulatory approval, and all participants provided informed consent. Significance of the percent changes from baseline was assessed using a mixed linear model; correlations were assessed with Pearson correlation. Nineteen (mean age, 30.4 years; 10 male/9 female) of the 23 patients who completed the pivotal trial enrolled in the extension trial, and 17 completed week 126 (78 weeks pivotal + 48 weeks extension) assessments (primary efficacy end point). Three patients discontinued prematurely (relocation, elevated transaminases and excess alcohol, sudden cardiac death). The median lomitapide dose remained mostly consistent at 40 mg (range, 20–60 mg) from week 36 in the pivotal study to week 282 in the extension trial. Overall, the median treatment duration with lomitapide across both trials was 5.1 years (range, 2.1–5.7 years). Among the 17 patients who completed week 126, LDL-C decreased from 356±127 mg/dL at baseline to …