Hyperlipidemic Subjects Research Articles

Evaluation of plasma phytosterols in sitosterolemia, their kindreds and hyperlipidemia subjects

BackgroundPatients suffering from sitosterolemia with ABCG5/8 mutation typically present with early-onset or rapidly progressive atherosclerosis. Their kindreds with partial genetic deficiencies of ABCG5/8 are often considered healthy. However, discerning sitosterolemia from its familial kindreds and hyperlipidemia subjects has remained challenging. MethodsHere we retrospectively recruited seven families including 8 individuals diagnosed with sitosterolemia subjects, and 14 kindreds carrying single gene mutations. Additionally, 17 individuals with hyperlipidemia and 130 healthy controls served as positive and negative controls, respectively. A total of 6 phytosterols combined with cholesterol absorption indices (including sitosterol, campesterol, stigmasterol, and cholestanol) and cholesterol synthesis markers (desmosterol and 7-dehydrocholesterol), was compared across the aforementioned four groups. ResultsAs expected, the sitosterolemia subjects with double mutations demonstrated significantly elevated levels of sitosterol and other cholesterol absorption indices. Meanwhile, sitosterolemia kindreds with single gene mutation showed a similar pattern of activated cholesterol-absorption ability to the hyperlipidemia group, but not as high as the double mutation group. Notably, the cholesterol-synthesis enzyme 7-dehydrocholesterol reductase displayed an increase in the hyperlipidemia group but a decrease in the sitosterolemia kindred group, suggesting a potential discriminative role of 7-dehydrocholesterol in distinguishing between these two groups. The combination of phytosterols was more valuable than clinical lipid index for sitosterolemia diagnosis. ConclusionOur study revealed mild disruptions of cholesterol absorption capacities in sitosterolemia kindreds with single mutations. Furthermore, the combination of 6 phytosterols proved effective in distinguishing between sitosterolemia, its single mutation carriers, and hyperlipidemia patients.

Elevated Kallistatin promotes the occurrence and progression of non-alcoholic fatty liver disease

Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease worldwide, and the development of non-alcoholic steatohepatitis (NASH) might cause irreversible hepatic damage. Hyperlipidemia (HLP) is the leading risk factor for NAFLD. This study aims to illuminate the causative contributor and potential mechanism of Kallistatin (KAL) mediating HLP to NAFLD. 221 healthy control and 253 HLP subjects, 62 healthy control and 44 NAFLD subjects were enrolled. The plasma KAL was significantly elevated in HLP subjects, especially in hypertriglyceridemia (HTG) subjects, and positively correlated with liver injury. Further, KAL levels of NAFLD patients were significantly up-regulated. KAL transgenic mice induced hepatic steatosis, inflammation, and fibrosis with time and accelerated inflammation development in high-fat diet (HFD) mice. In contrast, KAL knockout ameliorated steatosis and inflammation in high-fructose diet (HFruD) and methionine and choline-deficient (MCD) diet-induced NAFLD rats. Mechanistically, KAL induced hepatic steatosis and NASH by down-regulating adipose triglyceride lipase (ATGL) and comparative gene identification 58 (CGI-58) by LRP6/Gɑs/PKA/GSK3β pathway through down-regulating peroxisome proliferator-activated receptor γ (PPARγ) and up-regulating kruppel-like factor four (KLF4), respectively. CGI-58 is bound to NF-κB p65 in the cytoplasm, and diminishing CGI-58 facilitated p65 nuclear translocation and TNFα induction. Meanwhile, hepatic CGI-58-overexpress reverses NASH in KAL transgenic mice. Further, free fatty acids up-regulated KAL against thyroid hormone in hepatocytes. Moreover, Fenofibrate, one triglyceride-lowering drug, could reverse hepatic steatosis by down-regulating KAL. These results demonstrate that elevated KAL plays a crucial role in the development of HLP to NAFLD and may be served as a potential preventive and therapeutic target.

Borderline hyperlipidemia preventive management with Berberine PL in asymptomatic prevention of early atherosclerosis.

The aim of this pilot, efficacy supplement registry was to use a supplementary management with berberine to control hyperlipidemia. The supplement Berberine (Berbevis™ as Sophy® tablets) was used to control lipids and to evaluate (as a natural, preventive management) the early evolution of subclinical atherosclerosis in subjects (otherwise healthy, not using drugs) with borderline hyperlipidemia. The registry involved two groups of subjects not using drugs for a total of 50 subjects and three months of supplementation. The registry groups using standard management (SM) or SM and supplement were resulted comparable. No side effects were observed during the three months of berberine supplementation. No tolerability problems were reported. All subjects managed with berberine completed the three-month registry. Compliance was >97% (% of correctly used tablets). Total cholesterol was significantly decreased with berberine (P<0.05) and HDL was significantly improved (P<0.5) with supplementation. Triglycerides decreased in the berberine groups (P<0.05) and the levels of CoQ10 remained within normal values in supplemented subjects. Oxidative stress - measured in Carr units - was significantly decreased with berberine (P<0.05). Routine blood tests remained within normal values during the registry. Body weight was significantly more decreased (P<0.05) with berberine in comparison with standard management. The fat proportion also decreased (P<0.05) with berberine supplementation and the abdominal fat thickness (in the peri-umbilical area) was significantly decreased after berberine supplementation (P<0.05). This pilot registry indicates that berberine administration is effective in reducing lipids (decreasing weight, fat percentage and abdominal fat) in otherwise healthy subjects not using drugs. A longer study, with more advanced hyperlipidemic subjects is suggested. Predictive analytics according to Siegel suggests that a six-month study with 60 patients, in more advanced hyperlipidemic, also evaluating the intima-media thickness for the analysis of vascular benefits, may produce a stronger evaluation for this product.

The effect of date seed powder supplementation on glycaemic markers and lipid profile in women with overweight and obesity: a randomised controlled trial

SummaryThis randomised controlled trial aimed to investigate the effect of date kernel on glycaemic control, lipid profile, and weight in women with overweight and obesity. A total of 124 women with overweight or obesity were randomly assigned to receive the date kernel supplement or placebo for 12 weeks. Haemoglobin A1c (HbA1c) was measured by high‐performance liquid chromatography and fasting plasma glucose and lipid profiles by enzyme‐linked immunosorbent assay kits. In all, 98 subjects aged 40.7 years completed the trial (48 in the date kernel group and 50 in the placebo group). After 12 weeks, date kernel supplementation resulted in a significant decrease in HbA1c [mean (SD): −0.45 (0.36)] and fasting plasma glucose compared to the placebo [−0.16 (0.27)]. No significant change was observed in the lipid profiles of the total participants. However, date kernel decreased cholesterol and triglyceride (TG) in hypercholesterolaemic and hypertriglyceridaemic participants compared to the placebo. Date kernel improved glycaemic markers in overweight or obese women compared with the placebo. Furthermore, a significant reduction in total cholesterol and TG was observed only in hyperlipidaemic subjects. Date seed is a source of valuable components, including dietary fibre and bioactive compounds, that could provide economic, nutritional, and health benefits to the nutraceutical and food industries.

Elevated Circulating Endocan Levels Are Associated with Increased Levels of Endothelial and Inflammation Factors in Postprandial Lipemia.

Postprandial lipemia (PPL) causes endothelial dysfunction by causing endothelial damage to lipoproteins that remain rich in triglycerides. Endocan is a proteoglycan with increased tissue expression, endothelial activation, and neovascularization. The aim of the study was to examine circulating endocan levels in PPL subjects by considering the degree of PPL response according to a high-fat test meal. The other aim was to determine the association between endocan levels and endothelial and inflammatory factors. Fifty-four hyperlipidemic subjects and 28 normolipidemic subjects consumed the high-fat meal. Endocan, sICAM-1, sVCAM-1, and VEGFA as endothelial factors and IL-6 and LFA-1α as inflammatory factors were evaluated. Fasting serum endocan, VEGFA, sICAM-1, sVCAM-1 IL-6, and LFA-1α levels were increased in the PPL group compared to the control group. The PPL group was divided into tertiles based on mean AUC levels. Endocan levels in tertile 3 were at the highest and were increased significantly compared to tertiles 1 and 2. AUC and endocan levels were positively correlated with other endothelial and inflammation factors. ROC analysis showed endocan levels to be one of the highest values. Circulating endocan is seen at significantly higher levels and independently associated with endothelial and inflammatory factors in postprandial lipemia and dyslipidemia.

Intestinal fat absorption shifting: polyglucosamine biopolymer controls lipids and weight and reduces the progression of subclinical atherosclerosis.

Intestinal fat absorption shifting (IFAS) can be obtained in hyperlipidemic subjects with polyglucosamine biopolymer (BP) able to segregate most metabolic fats in the gut, making them unavailable for intestinal interaction (shift). The aim of this study was to evaluate the effects of a SM (standard management) for hyperlipidemia in asymptomatic subjects for primary cardiovascular prevention focusing on arterial wall morphology (IMT thickness) in comparison to SM associated to the administration of the BP. Two groups of comparable subjects (SM and SM+oral BP, 3 g/day) were considered; subjects were managed - in a supplement, pilot registry - for a year. Weight, fat mass, lipid profile, oxidative stress, IMT (carotids), the presence of granulations at the internal arterial layers and "near wall low density 'bubbles' were observed and compared at 1 year of management. A non-parallel, comparable group of subjects (102) using a statin for the same conditions was used as a reference population. Two hundred eighty-four subjects completed one year (140 in the SM group and 144 in the SM+BP group). Compliance was optimal with (96.3% of the table correctly used) with no side effects. BMI, fat mass and oxidative stress decreased more in the SM+BP group (P<0.05). Cholesterol and triglycerides levels were significantly improved with BP (P<0.05). IMT measurements were significantly decreased (P<0.05) in the SM+BP group (as for the intimal granulation/bubbles) with minimal variations in the comparative SM group. In the statin group, the lipid profile was modified (P<0.05) but not the IMT and the rate of drop outs was higher (15.7%); these patients stopped the management; in 23% of these subjects muscular pain not seen with BP, was observed. These results indicate positive effects of IFAS due to BP on IMT and arterial wall morphology and weight after 12 months. Fat shifting at intestinal level and the reduction of oxidative stress limit lipid oxidation/deposition into the arterial wall.

Studies on life style and dietary pattern of seleceted hyper lipidemic subjects of Parbhani city in Maharashtra

Hypercholesterolemia or hyperlipidemia or dyslipidaemia, an anothernoncommunicable health problem. Hence, study was conducted with an objective to determine the socioeconomic status, information about dietary intake of hyperlipidemia. Thirty non-insulin dependent hyperlipidemic subjects (men and women) were selected from Parbhani city of Maharashtra state. Information on diet pattern, lifestyle, known risk factors for disease were collected.The information regarding their dietary habits, frequency of consumption of different food groups in a day etc. were collected.The actual food intake of the selected diabetic subjects was collected for the immediate past 24 hours. The intake of different nutrients per day by each selected subject was then calculated from the food intake values using food composition tables. Food and nutrient adequacy was calculated based on balanced diets and Recommended Dietary Allowances respectively.It was observed that from hyperlipidemic subjects were more from 50-60 years age group (76.66 %) respectively. Sex wise data indicated that more number of male were sufferers of hyperlipidemia (70 %).Hyperlipidemia was observed more among the subjects who were engaged in the service (36.66 % each). The income wise distribution indicated that there was increase in the number of subjects as the income was increased. A very high per cent (70%) of males were having hyperlipidemia.There were 6.66 per cent and 13.33 per cent subjects who had impaired lipid profile form 3-5 years and &gt; 5 years duration.The increase in weight was experienced by all the selected 30 hyperlipidemic subjects after diagnosis of disorder.There were more than half with family history of hyperlipidemia. Oilseeds were consumed daily by 60 per cent hyperlipidemic respondents. It can be concluded that hyperlipidemia can be well controlled by modification in the diet and lifestyle.

Safety and efficacy of oil palm phenolic supplementation in improving lipid profile among hyperlipidemic adults: a phase 2, randomized, double-blind, placebo-controlled clinical trial.

Introduction: Oil palm phenolic (OPP) is an antioxidant aqueous palm oil by-product and contains a high amount of phenolics. OPP has been proven to have many therapeutical benefits, and one of them is as an antihyperlipidemic agent. The previous phase 1 clinical trial proved OPP was safe to be orally consumed by healthy volunteers and yielded a good lipid profile. Thus, this phase 2 clinical trial was conducted to determine the effectiveness of OPP in improving the lipid profile among hyperlipidemic subjects. Methods: A parallel, placebo-controlled, randomized, double-blinded clinical trial was conducted for 2months on 50 hyperlipidemic subjects aged 20-50years old. The subjects were randomly distributed to two treatment arms with 25 participants each: control/placebo (11 males and 14 females) and 250mg of OPP (10 males and 15 females). The subjects were required to consume one capsule per day for 60days. Fasting blood sampling for routine blood profile (hematology, liver function, renal function, and lipid) analysis and a medical examination were conducted at baseline, day 30, and day 60. t-test analysis was used to compare the difference between two test groups. Results: The baseline lipid profile between control group (TC, 5.78 ± 0.52mmol/L; LDL, 3.88 ± 0.51mmol/L; HDL, 1.30 ± 0.25; TG, 1.30 ± 0.82), and 250mg OPP (TC, 5.76 ± 0.54mmol/L; LDL, 3.82 ± 0.59mmol/L; HDL, 1.37 ± 0.34; TG, 1.25 ± 0.54) is insignificant. No serious adverse events (SAEs) were reported. No abnormality in fasting blood parameters in all groups was found. Compared to the control group among male participants, the 250mg OPP group showed an improved serum triglyceride level. There were no statistically significant changes in all blood parameters from day 1 to day 60 with the exception of triglyceride level. Conclusion: The absence of SAEs reported and no abnormal findings in biochemistry and hematology results suggested that the 250mg OPP was safe to be taken by hyperlipidemic patients with a high probability of reducing triglyceride level in hyperlipidemic male patients The outcomes from this phase II trial suggest that by incorporating OPP supplements into the diet may be a promising strategy for individuals with hyperlipidemia to improve their lipid profiles and reduce cardiovascular risk. However, more research is needed to fully understand the mechanisms of action and establish the long-term efficacy and safety of OPP supplementation in larger scale. Limitation: Small samples size hence lack of diversity (25 subjects per groups) and early sharing of treatment-response results. Clinical Trial Registration: clinicaltrials.gov, identifier NCT04573218.

Genetic polymorphism of fatty acid binding protein-2 in hyperlipidemic Asian Indians in North India.

Fatty acid binding protein-2 (FABP-2) is involved in the metabolism of lipids in the intestine. FABP-2 Ala54Thr polymorphism involves a transition of G to A at codon 54 of FABP-2, resulting in an amino acid substitution Ala54 to Thr54 and is associated with elevated fasting triglycerides in some hyperlipidemic populations. In current genome builds and gene databases the variant of the Ala54Thr FABP-2 (rs 1 799 883) is annotated as c.163A>G (p. Thr55Ala). The status of this polymorphism in hyperlipidemic Asian Indians from North India has not been investigated. This study was aimed to evaluate the distribution of the polymorphic variants of the Ala54Thr FABP-2 and their association with lipids in hyperlipidemic subjects. Ala54Thr FABP-2 polymorphism in both hyperlipidemic (n=210) and normolipidemic (n=342) subjects was assessed by PCR-RFLP. Ala54Thr genotypes and alleles distribution did not differ between the hyperlipidemic and normolipidemic groups. The heterozygous genotype FABP-2 Ala/Thr was significantly associated with higher levels of triglycerides and very low-density lipoproteins as compared to the homozygous variant (Thr/Thr) genotype and the wild type homozygous (Ala/Ala) genotype. The heterozygous genotype FABP-2 Ala54Thr is a risk factor for the development of hypertriglyceridemia and increased levels of VLDL-c in Asian Indians from North India.

Prevalence of genetically defined familial hypercholesterolemia and the impact on acute myocardial infarction in Taiwanese population: A hospital-based study.

BackgroundFamilial hypercholesterolemia (FH) is a common genetic disorder with markedly increased risk of coronary artery diseases (CAD), especially acute myocardial infarction (AMI). However, genetic tests for FH are not always necessary in the current diagnostic criteria of FH, which might lead to underestimation of the prevalence of FH and a lack of awareness of FH-associated CAD and AMI. We aimed to explore the prevalence of genetically defined FH in the hospital-based population and to determine the impact of FH risk variants on CAD and AMI.MethodsThe study participants were recruited between June 24, 2019 and May 12, 2021, at a medical center in Taiwan, in cooperation with the Taiwan Precision Medicine Initiative (TPMI) project. The prevalence of FH was calculated and the effects of FH pathogenic variants on CAD and AMI were analyzed by logistic regression models and shown as ORs and 95% CI.ResultsThe prevalence of genetically defined FH was 1.13% in the hospital-based population in Taiwan. Highest LDL and total cholesterol levels were observed in patients with LDLR rs28942084 (LDL 219.4±55.2; total cholesterol 295.8±55.4). There was an approximately 4-fold increased risk of hyperlipidemia in subjects with the LDLR rs769446356 polymorphism (OR, 4.42; 95% CI, 1.92-10.19) and AMI in individuals with the LDLR rs730882109 polymorphism (OR, 3.79; 95% CI, 2.26-6.35), and a 2-fold increased risk of CAD in those with the LDLR rs749038326 polymorphism (OR, 2.14; 95% CI, 1.31-3.50), compared with the groups without pathogenic variants of FH.ConclusionsThe prevalence of genetically defined FH was 1.13% in the hospital-based population in Taiwan, which was higher than the rate observed in individuals with clinically defined FH. The risk of CAD and AMI was increased to varying degrees in subjects with different FH risk alleles. Close monitoring and risk stratification strategy are essential in high-risk patients with FH risk alleles to facilitate early detection and treatments.

The Pattern of Dyslipidemia and its Association with Dietary Habits in Individuals Attending BSMMU Outpatient Department

Introduction: Abnormal level of circulating lipids refers to dyslipidemia. Dyslipidemia mostly affects non-communicable diseases like CHD, CVD, cancer, autoimmune disease, etc. The population group most affected by non-communicable diseases (NCDs) in Bangladesh comprises middle-aged persons and the elderly, having a major share of the disease burden and mortality in the country. Changing dietary habits and lifestyle, rapid urbanization, growth of commuting, tobacco use, uncontrolled growth and consumption of processed foods and beverages, indoor air pollution, road-traffic injuries, lack of awareness about healthful behavioral patterns, and psychological pressure are among the important factors responsible for such non-communicable diseases. The present study was conducted with the goal of observing any association between dyslipidemia and dietary habits in men and women. Aim of the study: The aim of the study was to observe the association of dyslipidemia with dietary habits. Methods: This cross-sectional analytical study was conducted at the Department of Biochemistry, Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka, Bangladesh. The study duration was one year, from March 2016 to February 2017. Non-probability sampling method was used to select a total of 245 dyslipidemia cases from the adult population attending the BSMMU outpatient department during the study period. Result: The majority of the dyslipidemic patients had isolated hypercholesterolemia. All types of dyslipidemia had a significantly higher prevalence among the male population. Mixed hyperlipidemia was significantly higher among the older population. Statistical significance was only observed among the total cholesterol levels and the male population and TG level and the female population of mixed hyperlipidemic subjects in regards to healthy food consumption. Unhealthy food consumption had a high correlation with dyslipidemia in both male and female population. Conclusion: The study shows that isolated hypercholesterolemia is prevalent. Food habits also have an impact on lipid profile among study subjects. Healthy foods are mostly associated with high HDL-C. Unhealthy food consumption has a high correlation with dyslipidemia in both male and female populations.

EFFICACY OF A LIPID-LOWERING DIET ON KEY FATTY ACID RATIOS AND OMEGA-3 INDEX IN HYPERLIPIDEMIC SUBJECTS.

Polyunsaturated fatty acid (PUFA) dietary intake, status and serum key fatty acid (FA) ratios may aid in cardiovascular disease-related risk assessment. The aim of this study was to investigate the effects of lipid-lowering diet on key FA ratios in serum phospholipids and omega-3 index in erythrocyte phospholipids in moderately hyperlipidemic subjects. The study included 41 subjects, mean age 56±6 years. Nutritional habits were evaluated by food frequency questionnaire. Participants followed lipid lowering diet for 12 weeks. Energy intake of omega-6 and omega-3 FA was changed from 7.6% and 0.6% to 5.7% and 1.2%, respectively. Marked decrease in four FA ratios in serum phospholipids, i.e., omega-6/omega-3, arachidonic acid (AA)/eicosapentaenoic acid (EPA), AA/docosahexaenoic acid (DHA), AA/(EPA+DHA) and omega-3 index (EPA+DHA) was found in study subjects after lipid-lowering diet. Total cholesterol/high-density lipoprotein (HDL), low-density lipoprotein (LDL)/HDL and triacylglycerol/HDL-cholesterol ratios positively correlated with all FA ratios, and negatively correlated with total omega-3 levels in serum phospholipids and omega-3 index in erythrocytes. Total serum omega-3 levels showed strongest association with lipoprotein ratios and positive correlation with homeostatic model assessment (HOMA) index. In conclusion, lipid-lowering diet resulted in decreased serum key FA ratios, increased omega-3 levels, and improved insulin sensitivity that may lead to a lower risk of cardiovascular disease in subjects with moderate hyperlipidemia.

Assessment The effect of prebiotics, probiotics and synbiotics on Hyperlipidemia

Hypercholesterolemia is a major risk factor for the development of atherosclerosis; a number of pharmacological and non-pharmacological (including dietary) approaches being employed to reduce it. Up until now, the trials to prove the hypocholesterolemia effect of probiotics have not been convincing. There are only few data suggesting that the supplementation of the diet with functional food products containing probiotic bacteria lower LDL-cholesterol (LDL-C) concentration in patients with moderately elevated cholesterol concentration in plasma. The aim of the present study was to evaluate in vitro the anti-atherosclerotic effect of a probiotic (Lactobacillus rhamnosus, casei, plantarum, and reuteri), prebiotic (Flaxseed & Cinnamon, green coffee, ginger, and green tea extracts) & combination mix using MCF-7cell line. Recording data showed that probiotic compound administration to hyperlipidemia: a significant decrease of the total serum cholesterol & triglycerides, a significant increase of the serum antioxidant potential. In conclusion, our data support the administration of probiotic Lactobacillus bacteria and prebiotic and combination to decrease serum cholesterol, triglycerides, and lipid droplets increase the antioxidant potential in hyperlipidemia subjects.

Promoter polymorphism of TNF‑α (rs1800629) is associated with ischemic stroke susceptibility in a southern Thai population

Stroke represents the leading cause of disability and mortality amongst the elderly worldwide. Multiple risk factors, including both genetic and non-genetic components, as well as their interactions, are proposed as etiological factors involved in the development of ischemic stroke (IS). Promoter polymorphisms of the IL-6-174G/C (rs1800795) and TNF-α-308G/A (rs1800629) genes have been considered as predictive risk factors of IS; however, these have not yet been evaluated in a Thai population. The aims of this study were to investigate the association of IL-6-174G/C and TNF-α-308G/A polymorphisms with IS. Genomic DNA from 200 patients with IS and 200 controls were genotyped for IL-6-174G/C and TNF-α-308G/A polymorphisms using TaqMan™ SNP genotyping and quantitative PCR-high resolution melting analysis, respectively. It was found that the TNF-α-308 A allele was significantly associated with an increased risk of IS development compared with the G allele [odds ratio (OR)=2.044; 95% CI=1.154-3.620; P=0.014]. Moreover, the IS risk was significantly higher in the presence of TNF-α-308 GA or AA genotypes compared with that in the presence of GG genotypes with a dominant inheritance (OR=1.971; 95% CI=1.080-3.599; P=0.027). However, there was no association between IL-6-174G/C and the risk of IS development. The interaction study demonstrated that IL-6-174 GG and TNF-α-308 GG genotypes enhanced IS susceptibility when combined with hypertension, hyperlipidemia and alcohol consumption. Hypertensive and hyperlipidemic subjects with the TNF-α-308 GA and AA genotypes were more likely to develop IS compared with those who did not have these two conditions and had the GG genotype. In a matched study design (1:1), the IL-6-174 GC genotype was associated with higher IL-6 levels in the control group. Collectively, the present results highlight the utility of the TNF-α-308G/A polymorphism as a predictive genetic risk factor for development of IS.

The Effect of Temperature on the Stability of PCSK-9 Monoclonal Antibody: An Experimental Study

Background: PCSK9 monoclonal antibody lowers plasma PCSK9 and LDL-cholesterol levels. The manufacturers recommend drug storage at 2–8°C, and not above 25°C. This study aimed to investigate drug stability at various temperatures that this drug could be exposed to during medication handling and transportation in tropical countries. Methods: Alirocumab and evolocumab were tested in 3 study conditions: room temperature (RT), cooler device with cold pack, and freeze-thaw for 9 and 18 hours. Heated drugs were used as negative control. Free plasma PCSK9 levels from 9 hyperlipidemia subjects were measured with ELISA. Results: Average subject age was 49.2±18.4 years. Percent PCSK9 inhibition significantly declined in heated drugs compared to baseline. Average RT during the study period was 30.4°C. Change in percent PCSK9 inhibition of PCSK9 mAb at RT from baseline was -5.8±4.4 (p=0.005) and -11.0±8.9% (p=0.006) for alirocumab at 9 hours and 18 hours, and -9.7±11.8% (p=0.04) and -15.1±14.3% (p=0.01) for evolocumab at 9 and 18 hours, respectively. In contrast, there were no significant changes in percent PCSK9 inhibition from baseline when PCSK9 mAb was stored in a cooler. In freeze-thaw condition, changes in percent PCSK9 inhibition from baseline to 9 and 18 hours were -5.2±2.9% (p=0.001) and -2.6±4.9% (p=0.16) for alirocumab, and -1.8±4.2% (p=0.24) and 0.4±6.1% (p=0.83) for evolocumab. Conclusion: Proper drug storage according to manufacturer’s recommendation is essential. Drug storage at RT in tropical climate for longer than 9 hours significantly decreased drug efficacy; however, storage in a cooler device with cold pack for up to 18 hours is safe.

Decreased Efficiency of Very-Low-Density Lipoprotein Lipolysis Is Linked to Both Hypertriglyceridemia and Hypercholesterolemia, but It Can Be Counteracted by High-Density Lipoprotein.

Impaired triglyceride-rich lipoprotein plasma catabolism is considered the most important factor for hypertriglyceridemia development. The aim of this study was to evaluate the impact of hypercholesterolemia and hypertriglyceridemia on the efficiency of lipoprotein lipase (LPL)-mediated very-low-density lipoprotein (VLDL)-triglyceride lipolysis and the role of high-density lipoprotein (HDL) in this process. Subjects with no history of cardiovascular disease (CVD) and untreated with lipid-lowering agents were recruited into the study and divided into normolipidemic, hypercholesterolemic, and hyperlipidemic groups. VLDL was isolated from serum and incubated with LPL in the absence or presence of HDL. For the hypercholesterolemic and hyperlipidemic groups, a significantly lower percentage of hydrolyzed VLDL-triglyceride was achieved compared to the normolipidemic group (p < 0.01). HDL enhanced the lipolysis efficiency in the hypercholesterolemic and hyperlipidemic groups on average by ~7% (p < 0.001). The lowest electrophoretic mobility of the VLDL remnants indicating the most effective lipolysis was obtained in the normolipidemic group (p < 0.05). HDL presence significantly reduced the electrophoretic mobility of the VLDL remnants for the hypercholesterolemic and hyperlipidemic groups (p < 0.05). The results of our study indicate that VLDL obtained from hypercholesterolemic and hyperlipidemic subjects are more resistant to lipolysis and are additional evidence of the need for early implementation of hypocholesterolemic treatment, already in asymptomatic CVD subjects.

Gamma-glutamyl carboxylated Gas6 facilitates the prophylactic effect of vitamin K in inhibiting hyperlipidemia-associated inflammatory pathophysiology via arresting MCP-1/ICAM-1 mediated monocyte-hepatocyte adhesion

Role of growth arrest-specific 6 (Gas6), member of vitamin K (VK)-dependent protein family in hyperlipidemia-associated inflammation remains unresolved. To address this, blood samples were collected from hyperlipidemic subjects and age-matched healthy controls and observed that gamma-glutamyl carboxylated Gas6 (Gla-Gas6) but not total Gas6 were significantly lower while pro-inflammatory markers, MCP-1 and ICAM-1 were remarkably higher in hyperlipidemic subjects compared to control. Correlation analyses demonstrated that Gla-Gas6 levels were inversely correlated with MCP-1 and ICAM-1 but positively with plasma VK in hyperlipidemic subjects but not in control. This suggests that boosting VK level might ameliorate the hyperlipidemia-associated inflammatory pathophysiology via augmenting Gla-Gas6. Further studies with high fat diet (HFD)-fed mice demonstrated that VK supplementation (1, 3, and 5 µg/kg BW, 8 weeks) dose-dependently reduced both hepatic and plasma levels of MCP-1 and ICAM-1 while elevating that of Gla-Gas6 but not total Gas6 in HFD-fed mice. Cell culture studies with gamma-glutamyl carboxylase (enzyme causes VK-dependent carboxylation of Gas6) knockdown hepatocytes and monocytes dissected the direct role of Gla-Gas6 in inhibiting high palmitic acid (0.75 mM)-induced inflammation via arresting MCP-1/ICAM-1 mediated hepatocyte-monocyte adhesion. The present study demonstrated an important role of Gla-Gas6 in facilitating the prophylactic effect of VK against hyperlipidemia associated inflammation.

Machine learning modelling of blood lipid biomarkers in familial hypercholesterolaemia versus polygenic/environmental dyslipidaemia

Familial hypercholesterolaemia increases circulating LDL-C levels and leads to premature cardiovascular disease when undiagnosed or untreated. Current guidelines support genetic testing in patients complying with clinical diagnostic criteria and cascade screening of their family members. However, most of hyperlipidaemic subjects do not present pathogenic variants in the known disease genes, and most likely suffer from polygenic hypercholesterolaemia, which translates into a relatively low yield of genetic screening programs. This study aims to identify new biomarkers and develop new approaches to improve the identification of individuals carrying monogenic causative variants. Using a machine-learning approach in a paediatric dataset of individuals, tested for disease causative genes and with an extended lipid profile, we developed new models able to classify familial hypercholesterolaemia patients with a much higher specificity than currently used methods. The best performing models incorporated parameters absent from the most common FH clinical criteria, namely apoB/apoA-I, TG/apoB and LDL1. These parameters were found to contribute to an improved identification of monogenic individuals. Furthermore, models using only TC and LDL-C levels presented a higher specificity of classification when compared to simple cut-offs. Our results can be applied towards the improvement of the yield of genetic screening programs and corresponding costs.

A Study of Blood Fatty Acids Profile in Hyperlipidemic and Normolipidemic Subjects in Association with Common PNPLA3 and ABCB1 Polymorphisms.

Adiponutrin (patatin-like phospholipase domain-containing 3; PNPLA3), encoded in humans by the PNPLA3 gene, is a protein associated with lipid droplet and endoplasmic reticulum membranes, where it is apparently involved in fatty acid redistribution between triglycerides and phospholipids. A common polymorphism of PNPLA3 (I148M, rs738409), linked to increased PNPLA3 presence on lipid droplets, is a strong genetic determinant of non-alcoholic fatty liver disease (NAFLD) and of its progression. P-glycoprotein (Pgp, MDR1—multidrug resistance protein 1, ABCB1—ATP-binding cassette sub-family B member 1), encoded by the ABCB1 gene, is another membrane protein implicated in lipid homeostasis and steatosis. In the past, common ABCB1 polymorphisms have been associated with the distribution of serum lipids but not with fatty acids (FA) profiles. Similarly, data on the effect of PNPLA3 I148M polymorphism on blood FAs are scarce. In this study, a gas chromatography-flame ionization detection (GC-FID) method was optimized, allowing us to analyze twenty FAs (C14: 0, C15: 0, C15: 1, C16: 0, C16: 1, C17: 0, C17: 1, C18: 0, C18: 1cis, C18: 2cis, C20: 0, C20: 1n9, C20: 2, C20: 3n6, C20: 4n6, C20: 5, C23: 0, C24: 0, C24: 1 and C22: 6) in whole blood, based on the indirect determination of the fatty acids methyl esters (FAMES), in 62 hyperlipidemic patients and 42 normolipidemic controls. FA concentrations were then compared between the different genotypes of the rs738409 and rs2032582 (ABCB1 G2677T) polymorphisms, within and between the hyperlipidemic and normolipidemic groups. The rs738409 polymorphism appears to exert a significant effect on the distribution of blood fatty acids, in a lipidemic and fatty acid saturation state-depending manner. The effect of rs2032582 was less pronounced, but the polymorphism did appear to affect the relative distribution of blood fatty acids between hyperlipidemic patients and normolipidemic controls.

Abstract 12579: CVI-LM001, a First-in-class Novel Oral PCSK9 Modulator, Lowers Plasma Ldl-c and Reduces Circulating PCSK9 in Preclinical Animal Models and in Hyperlipidemic Human Subjects

Hypercholesterolemia is a major risk factor for atherosclerotic cardiovascular disease (ASCVD) and lowering low-density lipoprotein cholesterol (LDL-C) levels via upregulation of hepatic LDL receptor (LDLR) produces CVD benefits. More recently, monoclonal antibodies targeting PCSK9, a LDLR degradation protein, has emerged as a new therapeutic approach for lowering LDL-C levels, offering great efficacy and positive benefits in ASCVD patients. However, PCSK9 inhibitors require subcutaneous injections and the cost of therapy is high. CVI Pharmaceuticals has discovered a series of novel PCSK9 small molecule modulators that can reduce PCSK9 gene expression and increase LDLR abundance by PCSK9 dependent and independent mechanisms in hepatocytes. Among them, the lead compound CVI-LM001 is advanced to Phase 2 stage to treat hypercholesterolemic patients. Here we report that in hyperlipidemic hamsters, treatment with CVI-LM001 (40, 80 and 160 mg/kg, QD) for 4 weeks dose-dependently increased liver LDLR protein levels up to 3.5-fold and decreased circulating PCSK9 levels to 10% of control at the highest dose, this was accompanied by significant reductions in serum LDL-C. In a double-blind, randomized Phase 1a study conducted in healthy volunteers with normal lipid levels, compared to baseline, we observed a 36.4% (p&lt;0.001) reduction in serum PCSK9 levels after 10 days of oral treatment with CVI-LM001 (300 mg, QD). Moreover, in a Proof of Mechanism Phase 1b study conducted in subjects with elevated LDL-C, compared with placebo cohort, treatment with CVI-LM001 (300 mg, QD) for 28 days significantly reduced serum LDL-C (-26.3% p&lt;0.01), TC (-20.1%, p&lt;0.01), Apo B (-17.4%, p=0.01) and PCSK9 (-39.2%, p&lt;0.05). CVI-LM001 had a benign safety profile and was well tolerated in 105 treated healthy volunteers and 33 treated hyperlipidemic subjects. In addition, CVI-LM001 exhibited excellent pharmacokinetics properties with peak concentrations occurred approximately 1-1.5h post-dose. Mean half-lives ranged from 32 to 45h after single dosing and 62 to 68h following multiple dosing. These studies, for the first time, demonstrate that CVI-LM001, a novel PCSK9 modulator, has the potential to be a new oral cholesterol-lowering drug and warrant further development.