Margherita Botta

Circulating Levels of Proprotein Convertase Subtilisin/Kexin Type 9 and Arterial Stiffness in a Large Population Sample: Data From the Brisighella Heart Study

Background Proprotein convertase subtilisin/kexin type 9 (PCSK9) circulating levels are significantly associated with an increased risk of cardiovascular events. This study aimed to evaluate the relationship between circulating levels of PCSK9 and arterial stiffness, an early instrumental biomarker of cardiovascular disease risk, in a large sample of overall healthy participants. Methods and Results From the historical cohort of the Brisighella Heart Study, after exclusion of active smokers, participants in secondary prevention for cardiovascular disease, and patients in treatment with statins or vasodilating agents, we selected 227 premenopausal women and 193 age‐matched men and 460 postmenopausal women and 416 age‐matched men. In these participants, we evaluated the correlation between PCSK9 plasma circulating levels and pulse wave velocity. Postmenopausal women showed higher PCSK9 levels (309.9±84.1 ng/mL) compared with the other groups (P<0.001). Older men had significant higher levels than younger men (283.2±75.6 versus 260.9±80.4 ng/mL; P=0.008). In the whole sample, pulse wave velocity was predicted mainly by age (B=0.116, 95% CI 0.96–0.127, P<0.001), PCSK9 (B=0.014, 95% CI 0.011–0.016, P<0.001), and serum uric acid (B=0.313, 95% CI 0.024–0.391, P=0.026). Physical activity, low‐density lipoprotein cholesterol, high‐density lipoprotein cholesterol, and estimated glomerular filtration rate were not associated with pulse wave velocity (P>0.05).By considering the subgroups described, age and PCSK9 levels were mainly associated with pulse wave velocity, which also correlated with serum uric acid in postmenopausal women. Conclusions In the Brisighella Heart Study cohort, circulating PCSK9 is significantly related to arterial stiffness, independent of sex and menopausal status in women.

PPAR Agonists and Metabolic Syndrome: An Established Role?

Therapeutic approaches to metabolic syndrome (MetS) are numerous and may target lipoproteins, blood pressure or anthropometric indices. Peroxisome proliferator-activated receptors (PPARs) are involved in the metabolic regulation of lipid and lipoprotein levels, i.e., triglycerides (TGs), blood glucose, and abdominal adiposity. PPARs may be classified into the α, β/δ and γ subtypes. The PPAR-α agonists, mainly fibrates (including newer molecules such as pemafibrate) and omega-3 fatty acids, are powerful TG-lowering agents. They mainly affect TG catabolism and, particularly with fibrates, raise the levels of high-density lipoprotein cholesterol (HDL-C). PPAR-γ agonists, mainly glitazones, show a smaller activity on TGs but are powerful glucose-lowering agents. Newer PPAR-α/δ agonists, e.g., elafibranor, have been designed to achieve single drugs with TG-lowering and HDL-C-raising effects, in addition to the insulin-sensitizing and antihyperglycemic effects of glitazones. They also hold promise for the treatment of non-alcoholic fatty liver disease (NAFLD) which is closely associated with the MetS. The PPAR system thus offers an important hope in the management of atherogenic dyslipidemias, although concerns regarding potential adverse events such as the rise of plasma creatinine, gallstone formation, drug–drug interactions (i.e., gemfibrozil) and myopathy should also be acknowledged.

Plasma PCSK9 levels and lipoprotein distribution are preserved in carriers of genetic HDL disorders

Proprotein convertase subtilisin/kexin 9 (PCSK9), a protein regulating the number of cell-surface LDL receptors (LDLR), circulates partially associated to plasma lipoproteins. How this interaction alters PCSK9 plasma levels is still unclear. In the present study, we took advantage of the availability of a large cohort of carriers of genetic HDL disorders to evaluate how HDL defects affect plasma PCSK9 levels and its distribution among lipoproteins. Plasma PCSK9 concentrations were determined by ELISA in carriers of mutations in LCAT, ABCA1, or APOAI genes, and lipoprotein distribution was analyzed by FPLC. Carriers of one or two mutations in the LCAT gene show plasma PCSK9 levels comparable to that of unaffected family controls (homozygotes, 159.4u202fng/mL (124.9;243.3); heterozygotes, 180.3u202fng/mL (127.6;251.5) and controls, 190.4u202fng/mL (146.7;264.4); P for trendu202f=u202f0.33). Measurement of PCSK9 in plasma of subjects carrying mutations in ABCA1 or APOAI genes confirmed normal values. When fractionated by FPLC, PCSK9 peaked in a region between LDL and HDL in control subjects. In carriers of all HDL defects, lipoprotein profile shows a strong reduction of HDL, but the distribution of PCSK9 was superimposable to that of controls. In conclusion, the present study demonstrates that in genetically determined low HDL states plasma PCSK9 concentrations and lipoprotein distribution are preserved, thus suggesting that HDL may not be involved in PCSK9 transport in plasma.

High Density Lipoproteins Inhibit Oxidative Stress-Induced Prostate Cancer Cell Proliferation

Recent evidence suggests that oxidative stress can play a role in the pathogenesis and the progression of prostate cancer (PCa). Reactive oxygen species (ROS) generation is higher in PCa cells compared to normal prostate epithelial cells and this increase is proportional to the aggressiveness of the phenotype. Since high density lipoproteins (HDL) are known to exert antioxidant activities, their ability to reduce ROS levels and the consequent impact on cell proliferation was tested in normal and PCa cell lines. HDL significantly reduced basal and H2O2-induced oxidative stress in normal, androgen receptor (AR)-positive and AR-null PCa cell lines. AR, scavenger receptor BI and ATP binding cassette G1 transporter were not involved. In addition, HDL completely blunted H2O2-induced increase of cell proliferation, through their capacity to prevent the H2O2-induced shift of cell cycle distribution from G0/G1 towards G2/M phase. Synthetic HDL, made of the two main components of plasma-derived HDL (apoA-I and phosphatidylcholine) and which are under clinical development as anti-atherosclerotic agents, retained the ability of HDL to inhibit ROS production in PCa cells. Collectively, HDL antioxidant activity limits cell proliferation induced by ROS in AR-positive and AR-null PCa cell lines, thus supporting a possible role of HDL against PCa progression.

Influence of body variables in the development of metabolic syndrome—A long term follow-up study

Objectives The body variable associated with the diagnosis of Metabolic Syndrome (MetS) is an elevated waist circumference (WC), although a number of other variables have been suggested. Among these, an elevated waist-to-height ratio (WHtR), ie a value higher than 0.5, that may identify abnormality, independently from height. An elevated WHtR provided the best correlation with MetS in a prior study in a large Italian population. In order to assess the validity of this conclusion, a long-term follow-up study re-examined this population, also in order to detect possible associations with cardiovascular (CV) risk. Methods and results 1,071 subjects with a complete follow-up of over 6 years were evaluated with a comparative assessment of the three anthropometric variables, namely WHtR, WC and body mass index (BMI). WHtR≥ 0.5 had the highest sensitivity for the identification of MetS, both in males and females (94.1% and 86.7% respectively). WHtR was of reduced specificity, occurring, yet less frequently (17.7% in males and 30% in females), in patients without MetS. By contrast, enlarged WC occurred with a lower frequency in male patients who developed MetS (30.2%) whereas in females, frequency was higher than in males (69.3%). Finally, a BMI≥ 25 kg/m2 had intermediate sensitivity and specificity regardless of gender. WC showed the highest odds ratio (2.62, 95%CI: 1.18–5.78) for the prediction of CV occurrence. Conclusion The present study confirms WHtR as an excellent screening tool in identifying MetS carriers, but, different from reports in other countries, it shows a lower specificity in our population.

From lipoprotein apheresis to proprotein convertase subtilisin/kexin type 9 inhibitors: Impact on low-density lipoprotein cholesterol and C-reactive protein levels in cardiovascular disease patients:

In this observational study, we compared the effect of lipoprotein apheresis and evolocumab or alirocumab on levels of lipoprotein cholesterol, triglycerides and inflammatory markers (C reactive protein and interleukin 6) in cardiovascular patients (nu2009=u20099). Patients were monitored during the last year of lipoprotein apheresis followed by six months of treatment with proprotein convertase subtilisin/kexin type 9 inhibitors. The biochemical parameters were determined pre- and post- every apheresis procedure for 12 months and then after one, three and six months of treatment with evolocumab (140u2009mg every two weeks [Q2W]) or alirocumab (75u2009mg or 150u2009mg every two weeks [Q2W]). Lipoprotein apheresis significantly reduced low-density lipoprotein cholesterol levels from 138u2009±u200932u2009mg/dl to 46u2009±u200916u2009mg/dl (pu2009<u20090.001), with an inter-apheresis level of 114u2009±u200926u2009mg/dl. Lipoprotein(a) was also reduced from a median of 42u2009mg/dl to 17u2009mg/dl (pu2009<u20090.01). Upon anti-proprotein convertase subtilisin/kexin type 9 therapy, low-density lipoprotein cholesterol levels were similar to post-apheresis (59u2009±u200925, 41u2009±u200922 and 42u2009±u200921mg/dl at one, three and six months, respectively) as well as those of lipoprotein(a) (18u2009mg/dl). However, an opposite effect was observed on high-density lipoprotein cholesterol levels: –16.0% from pre- to post-apheresis and +34.0% between pre-apheresis and proprotein convertase subtilisin/kexin type 9 inhibitors. Apheresis significantly reduced high-sensitivity C-reactive protein levels (1.5u2009±u20091.2u2009mg/l pre-apheresis to 0.6u2009±u20090.6u2009mg/l post-apheresis), while no changes were found upon proprotein convertase subtilisin/kexin type 9u2009mAbs administration. In conclusion, our study demonstrated that, by switching from lipoprotein apheresis to anti-proprotein convertase subtilisin/kexin type 9 therapies, patients reached similar low-density lipoprotein cholesterol and lipoprotein(a) levels, increased those of high-density lipoprotein cholesterol, and showed no changes on high-sensitivity C-reactive protein.

PCSK9 deficiency reduces insulin secretion and promotes glucose intolerance: the role of the low-density lipoprotein receptor

AimsnPCSK9 loss of function genetic variants are associated with lower low-density lipoprotein cholesterol but also with higher plasma glucose levels and increased risk of Type 2 diabetes mellitus. Here, we investigated the molecular mechanisms underlying this association.nnnMethods and resultsnPcsk9 KO, WT, Pcsk9/Ldlr double KO (DKO), Ldlr KO, albumin AlbCre+/Pcsk9LoxP/LoxP (liver-selective Pcsk9 knock-out mice), and AlbCre-/Pcsk9LoxP/LoxP mice were used. GTT, ITT, insulin and C-peptide plasma levels, pancreas morphology, and cholesterol accumulation in pancreatic islets were studied in the different animal models. Glucose clearance was significantly impaired in Pcsk9 KO mice fed with a standard or a high-fat diet for 20u2009weeks compared with WT animals; insulin sensitivity, however, was not affected. A detailed analysis of pancreas morphology of Pcsk9 KO mice vs. controls revealed larger islets with increased accumulation of cholesteryl esters, paralleled by increased insulin intracellular levels and decreased plasma insulin, and C-peptide levels. This phenotype was completely reverted in Pcsk9/Ldlr DKO mice implying the low-density lipoprotein receptor (LDLR) as the proprotein convertase subtilisin/kexin Type 9 (PCSK9) target responsible for the phenotype observed. Further studies in albumin AlbCre+/Pcsk9LoxP/LoxP mice, which lack detectable circulating PCSK9, also showed a complete recovery of the phenotype, thus indicating that circulating, liver-derived PCSK9, the principal target of monoclonal antibodies, does not impact beta-cell function and insulin secretion.nnnConclusionnPCSK9 critically controls LDLR expression in pancreas perhaps contributing to the maintenance of a proper physiological balance to limit cholesterol overload in beta cells. This effect is independent of circulating PCSK9 and is probably related to locally produced PCSK9.

Efficacy and safety of a nutraceutical with probiotic and red yeast rice extract in patients with moderate hypercholesterolemia: A randomized, double-blind, placebo-controlled study

s / Atherosclerosis 263 (2017) e29ee110 e109 Aim: Our aim was to compare the ESC/EAS primary prevention guidelines with the idea of actual cardiovascular risk based on the results of carotid ultrasound. Methods: This cross-sectional study included 165 patients (53±8 years, 48% females) with at least one cardiovascular risk factor. Primary prevention strategy was identified based on global cardiovascular risk calculation by SCORE and the level of LDL-C. The strategy was compared to the findings of carotid ultrasound. The presence of plaque was defined as a marker of subclinical atherosclerosis. Results: According to the ESC/EAS guidelines, immediate drug intervention was required in 23 patients. In 18 of them the appropriateness of therapy was confirmed by the presence of plaque. Considering medical therapy after lifestyle modification should have been necessary in 106 participants and 34 of them had plaque. On the other hand, among 36 patients who didn’t require statins, 17% had carotid plaque. Thus, the calculated risk requiring statins for primary prevention had a sensitivity of 90% in prediction of carotid plaque presence. On the other hand, calculated risk not high enough to justify the drug intervention, predicted the absence of plaque with a specificity of 28%. Conclusions: The criteria for primary prevention based on the use of SCORE and LDL-C level are highly sensitive in detecting patients who have actually high risk identified by carotid imaging. On the other hand, the specificity of these guidelines may be lower, which results in identifying 72% of patients not having a carotid plaque as candidates for drug therapy for primary prevention. SAG221. EFFICACY AND SAFETY OF A NUTRACEUTICAL WITH PROBIOTIC AND RED YEAST RICE EXTRACT IN PATIENTS WITH MODERATE HYPERCHOLESTEROLEMIA: A RANDOMIZED, DOUBLE-BLIND, PLACEBO-CONTROLLED STUDY Chiara Macchi, Margherita Botta, Raffaella Bosisio, Chiara Pavanello, Chiara Maria Toldo, Giuliana Mombelli, Laura Calabresi, Massimiliano Ruscica, Paolo Magni. Dipartimento di Scienze Farmacologiche e Biomolecolari, Universit a degli Studi di Milano, Milano, Italy; Centro Grossi Paoletti, Dipartimento di Scienze Farmacologiche e Biomolecolari, Universit a degli Studi di Milano, Milano, Italy; Centro Dislipidemie, ASST Grande Ospedale Metropolitano Niguarda, Milano, Italy Aim: Aim. Specific probiotics, like the Bifidobacterium longum BB536, show a high biliary salt hydrolase activity and may contribute to lower circulating total cholesterol (TC) and LDL cholesterol (LDL-C) by reducing intestinal cholesterol reabsorption and may prove useful in the management of moderate dyslipidemia. Ă Study objective: to evaluate the effect of a nutraceutical mix (Bifidobacterium longum BB536, red yeast rice (RYR) extract (10 mg/day monacolin K), niacin, coenzyme Q10) on circulating lipid parameters (TC, HDLcholesterol (HDL-C), non-HDL-C, LDL-C, triglycerides (TG), apoliproteins AI (ApoAI) and B (ApoB), PCSK9) in subjects with borderline-high TC/LDL-C and low cardiovascular risk. Methods: Methods. Study design: randomized, double-blind, placebocontrolled study; intervention duration: 12 weeks. Thirtytwo subjects with moderate hypercholesterolemia (LDL-C: 130-200 mg/dL) were randomly allocated to either nutraceutical preparation or placebo. Results: Results. In active-treatment group, compared to placebo group, after 12 weeks, we observed a significant reduction of LDL-C (from 183 to 138 mg/dL (-25% vs. 0% (placebo)), TC (-16% vs. 0%), non-HDL-C (23% vs. 0%), ApoB (-20% vs. -1%). Similar changes were already achieved after 6 weeks. HDL-C, ApoAI, PCSK9 levels were unchanged and TG decreased to the same extent (-12/13%) in both arms. No adverse effects and a 97% compliance were observed. Conclusions: Conclusions: A 12-week treatment with a nutraceutical mix with the probiotic Bifidobacterium longum BB536, RYR extract, niacin and coenzyme Q10 was well tolerated by subjects with borderline hypercholesterolemia and resulted in a significant improvement of the proatherogenic lipid profile. ClinicalTrials.gov n NCT02689934. Supported by an unrestricted grant from Montefarmaco OTC S.p.A. (Bollate, MI, Italy). SAG222. ADHERENCE TO A MEDITERRANEAN DIET IS ASSOCIATED WITH THE PRESENCE AND EXTENT OF ATHEROSCLEROTIC PLAQUES IN MIDDLEAGED ASYMPTOMATIC ADULTS: THE AWHS STUDY Rocio Mateo Gallego, Irina Uzhova, Bel en Moreno-Franco, Montserrat Le on-Latre, Jos e A. Casasnovas, Fernando Civeira, Jos e L. Pe~ nalvo. Unidad Clínica y de Investigaci on en Lípidos y Arteriosclerosis, Hospital U. Miguel Servet, IIS Arag on, Zaragoza, Spain; 2 Fundaci on Centro Nacional de Investigaciones Cardiovasculares Carlos III (CNIC), Madrid, Spain; Unidad de Prevenci on Cardiovascular. Instituto Aragon es de Ciencias de la Salud (IACS), Zaragoza, Spain; Departamento de Medicina, Psiquiatría y Dermatología. Universidad de Zaragoza, Zaragoza, Spain; 5 Friedman School of Nutrition Science and Policy, Tufts University, Boston,