C. Giacobbe

Causative mutations and premature cardiovascular disease in patients with heterozygous familial hypercholesterolaemia

Background Familial hypercholesterolemia is a common autosomal dominant disease, caused by mutations leading to elevated low-density lipoprotein (LDL) cholesterol and, if untreated, to premature cardiovascular disease. Methods Patients (young adults with a family history of hypercholesterolaemia or premature cardiovascular disease) with LDL cholesterol concentration ≥4.9 mmol/l, after excluding Familial Combined Hyperlipidaemia, were evaluated for causative mutations, Dutch Lipid Clinic Network score calculation and non-invasive ultrasound examination of carotid arteries. Results Of the 263 patients, 210 were heterozygotes for LDL receptor (LDLR) mutations, four had APOB gene mutations, one PCSK9 gene mutation, while 48 had no evidence of mutations. Among 194 unrelated index cases 149 had mutations (77%). Among patients with LDLR mutations (n = 145), there were five compound heterozygotes, 75 patients with null mutations and 65 with missense mutations. As many as 178 patients underwent a follow-up and treatment (statin ± ezetimibe), achieving a mean reduction of 49% in LDL cholesterol, with 21% of patients reaching the LDL goal of 2.6 mmol/l. In a multivariate analysis, carotid plaques, at ultrasound examination, were associated with the presence of genetic mutation (p = 0.001), LDL cholesterol (p < 0.001), Dutch Lipid Clinic Network score (p < 0.001), independently of age, gender, smoking habits and systolic blood pressure. The presence of carotid plaque (p = 0.017), LDL cholesterol (p < 0.003), Dutch Lipid Clinic Network score (p < 0.001) were independently associated with premature cardiovascular disease. Conclusions We identified patients with causative mutations in 82% of the cases under study. In addition to LDL cholesterol and Dutch Lipid Clinic Network score, carotid plaques in ultrasound evaluation provide direct evidence of premature vascular disease and are associated with high risk for cardiovascular events.

Identification and in vitro characterization of two new PCSK9 Gain of Function variants found in patients with Familial Hypercholesterolemia

Familial hypercholesterolemia (FH) is an autosomal dominant disease caused by pathogenic variants in genes encoding for LDL receptor (LDLR), Apolipoprotein B and Proprotein convertase subtilisin/kexin type 9 (PCSK9). Among PCSK9 variants, only Gain-of- Function (GOF) variants lead to FH. Greater attention should be paid to the classification of variants as pathogenic. Two hundred sixty nine patients with a clinical suspect of FH were screened for variants in LDLR and the patients without pathogenic variants were screened for variants in PCSK9 and APOB. Functional characterization of PCSK9 variants was performed by assessment of protein secretion, of LDLR activity in presence of PCSK9 variant proteins as well as of the LDLR affinity of the PCSK9 variants. Among 81 patients without pathogenic variants in LDLR, 7 PCSK9 heterozygotes were found, 4 of whom were carriers of variants whose role in FH pathogenesis is still unknown. Functional characterization revealed that two variants (p.(Ser636Arg) and p.(Arg357Cys)) were GOF variants. In Conclusions, we demonstrated a GOF effect of 2 PCSK9 variants that can be considered as FH-causative variants. The study highlights the important role played by functional characterization in integrating diagnostic procedures when the pathogenicity of new variants has not been previously demonstrated.

EXPRESSION OF ADIPONECTIN RECEPTORS IN HUMAN CAROTID ATHEROSCLEROTIC PLAQUES Abstracts from the 12th National Congress of the Italian Society of Cardiovascular Prevention (SIPREC), Naples, 6–8 March 2014

Abstracts from the 12th National Congress of the Italian Society of Cardiovascular Prevention (SIPREC), Naples, 6–8 March 2014s from the 12th National Congress of the Italian Society of Cardiovascular Prevention (SIPREC), Naples, 6–8 March 2014 Springer International Publishing Switzerland 2014 LACTOBACILLUS PARACASEI B21060, ARABINOGALATTANO AND XILOOLIGOSACCHARIDES: SYMBIOTIC CANDIDATES TO REDUCE CHOLESTEROL LEVELS Ludovica Leone, Monica Malamisura, Antonia De Matteo, Valentina Bruno, Roberto Berni Canani, Basilio Malamisura Department of Translational Medicine, Pediatric Section and European Laboratory for the Investigation of Food Induced Diseases, University of Naples ‘‘Federico II’’, Via S. Pansini 5, 80131 Naples, Italy, Campus Biomedico University of Rome, Via Álvaro del Portillo 21, Rome, Italy, Department of Pediatrics, University Hospital of Salerno, Via San Leonardo, 1, 84131 Salerno, Italy Introduction: Elevated blood cholesterol is an important risk factor associated with atherosclerosis and coronary heart disease. Probiotics have been proposed for the treatment of dyslipidemia. Aim: To evaluate efficacy, tolerability and safety of a new symbiotic formulation containing a combination of probiotic (Lactobacillus paracasei B21060) and prebiotics (arabinogalattano, xilooligosaccharides) and amine (L-glutamine) in the early treatment of children affected by familial hypercholesterolemia (FH). Methods: FH subjects, aged 6–12 years, consecutively observed a Tertiary Center for Pediatric Nutrition were randomly allocated to two groups of intervention for 6 months: active group, received a low saturated fats diet plus the symbiotic (2.5 9 109 cfu, bid) for 6 months; control group, received low saturated fats diet alone. All children received written indications for low saturated fats diet. The plasmatic lipid profile was assessed by peripheral blood sampling at T0 and T1. Results: 40 FH children were enrolled (20 in active group: 8 male, median age 8.4 yrs, BMI 17.6; 20 in control group: 8 male, median age 7.5 years, BMI 17.0. All subjects completed the study. At T1 a reduction of C-LDL, total cholesterol, LDL/HDL ratio was observed in both groups, but the differences were significant only in active group (median value (IQR) C-LDL: 221 (55) vs 192 (33), p \ 0.01; total cholesterol: 280 (31) vs 260 (43), p \ 0.05; LDL/HDL: 4.0 (1.5) vs 3.5 (1.3), p \ 0.05). The symbiotic preparation was well accepted by the children, the adherence was [90 %, no side effects were observed. Conclusions: The symbiotic containing Lactobacillus paracasei B21060 is able to significantly reduce main laboratory biomarkers in children with FH. The treatment was well accepted and tolerated by patients. Our results open the light on new opportunity for the early treatment of pediatric FH. Further studies are advocated to better define the mechanism of action and the potential of a long term use of this new strategy. ANALYSIS OF THE INFLUENCE OF BASELINE DIABETES DURATION (DD) ON BLOOD PRESSURE RESPONSES TO LIRAGLUTIDE Angela Sciacqua, Francesco Perticone, Vivian Fonseca, Jorge Plutzky, Cristiano Bette, J. Hans DeVries Magna Graecia University, Catanzaro, Italy, Tulane University, New Orleans, LA, USA, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, USA, Novo Nordisk, Rome, Italy, Academic Medical Centre, University of Amsterdam, The Netherlands Introduction: Across the phase 3 ‘Liraglutide Effect and Action in Diabetes’ (LEAD)1–6 and liraglutide vs sitagliptin (Lira-DPP-4i) trials, once-daily liraglutide (LIRA) consistently reduced systolic blood pressure. In a sub-analysis of LEAD trials, significantly greater reductions in SBP were observed with LIRA in patients aged B65 vs [65 years. Since diabetes duration (DD) was 3–4 years longer in the groups [65 years, this High Blood Press Cardiovasc Prev (2014) 21:151–169 DOI 10.1007/s40292-014-0053-4