Serum PCSK9 Research Articles

Serum PCSK9 is a novel serological biomarker for the diagnosis and prognosis of pancreatic cancer.

Although CA19-9 is an essential blood biomarker of pancreatic cancer (PC), its sensitivity and specificity are limited for early detection. We analyzed the serum proprotein convertase subtilisin/kexin type 9 (sPCSK9) in PC patients, benign disease groups (BDG), and healthy controls (HC) by ELISA. Consistently, sPCSK9 was considerably lower in PC patients than in HC (Z = -2.546, P < 0.05), and sPCSK9 in PC patients was statistically significantly higher than in BDG (Z = -5.457, P < 0.001). sPCSK9 was linked to the invasion of lymph nodes (χ2 = 6.846, P < 0.01). According to ROC curves, combining sPCSK9 with CA19-9 could potentially enhance the diagnostic capability of CA19-9 in early-stage PC patients. Furthermore, the low sPCSK9 group (n = 41) exhibited statistically significantly prolonged overall survival compared to the high sPCSK9 group (n = 15), with median survival times of 27 months (95% CI [17.59-36.41]) and 11 months (95% CI [7.21-14.79]), respectively (P = 0.022). The diagnostic performance of CA19-9 for early-stage PC patients could be improved by combining sPCSK9 with CA19-9. Moreover, the higher sPCSK9 group has a significantly shorter overall survival rate.

Association between circulatory microRNA-218 expression, serum PCSK9 levels, inflammatory markers, and monocyte subsets in coronary artery disease patients: impact of statin therapy

BackgroundProprotein convertase subtilisin/kexin type-9 (PCSK9), an enzyme produced mainly by hepatocytes and breaks low-density lipoprotein receptor (LDL-R), inflammatory markers [toll like receptor 4 (TLR4), high mobility group box 1 (HMGB1), tumor necrosis factor alpha (TNFα), c-reactive protein (CRP)], and monocyte subtypes are associated with coronary artery disease (CAD) pathogenesis. The circulating microRNA-218 (miR-218) can relieve CAD through the suppression of HMGB1 in monocyte-derived inflammatory cytokines. Herein, we explored the association between circulatory miR-218 expression and serum levels of PCSK9, inflammatory markers, and monocyte subtypes in statin and non-statin CAD patients. This study involved 91 healthy (control) and 91 stable CAD participants which were subdivided into no-statin (NS, n = 25), low-statin (LS, n = 25), and high-statin (HS, n = 41) groups. low-density lipoprotein cholesterol (LDL-C) and CRP serum levels were calorimetrically determined. Serum levels of PCSK9, TLR4, HMGB1, and TNFα were detected by ELISA, while monocyte subsets [classical (CM), intermediate (IM), non-classical (NC)] were calculated by flow cytometry. Circulatory miR-218 expression was detected by real-time PCR.ResultsThe CAD group had significantly lower miR-218 expression and significantly higher levels of PCSK9, inflammatory markers (HMGB1, CRP, TLR4, and TNFα), and IM% than the control group. Among CAD patients, LS and HS groups had significantly lower miR-218 expression, LDL-C levels, and inflammatory markers and significantly higher levels of PCSK9 than the NS group. The HS group exhibited the lowest miR-218 expression and inflammatory markers and the highest PCSK9 levels. However, there were no significant changes in IM% among statin and non-statin groups. In the three CAD groups, miR-218 showed a significantly negative correlation with PCSK9 and inflammatory markers (HMGB1, CRP, TLR4, and TNFα), while this expression exhibited a significantly negative correlation with CM%, IM%, and NCM% only in the NS group. Results of multivariable linear regression indicated a correlation between miR-218 and five independent variables (PCSK9, HMGB1, CRP, TLR4, and TNFα) in the total statin (LS + HS) group, and eight independent variables (PCSK9, HMGB1, CRP, TLR4, and TNFα, CM%, IM%, NCM%) in the NS group. Provided that all other independent variables are constant, miR-218 expression was significantly correlated to CRP (Beta = 0.234) and PCSK9 (Beta = − 0.875) in the total statin group; TLR4 (Beta = − 0.554) in the LS group; HMGB1 (Beta = − 0.507) in the HS group; and CRP (Beta = − 0.745) in the NS group.ConclusionsStatin-treated CAD patients have a unique negative correlation between miR-218 and PCSK9, HMGB1, and TLR4, and subsequently with CAD progress. Therefore, it could be recommended to combine activators of miR-218 and inhibitors of PCSK9, HMGB1, and TLR4 with statin to efficiently treat CAD.

Serum pcsk9 levels as a novel biomarker for pregnant women with risk of fetal neural tube defects

Serum pcsk9 levels as a novel biomarker for pregnant women with risk of fetal neural tube defects

Mediation effect of metabolic factors and inflammation on the association of serum uric acid with serum proprotein convertase subtilisin/kexin type 9 levels

Mediation effect of metabolic factors and inflammation on the association of serum uric acid with serum proprotein convertase subtilisin/kexin type 9 levels

Association between serum PCSK9 and coronary heart disease in patients with type 2 diabetes mellitus

Background and aimsProprotein convertase subtilisin/kexin type 9 (PCSK9) is considered a new biomarker for atherosclerosis, but its ability to predict cardiovascular outcomes has been controversial. This study aimed to address the lack of data on PCSK9, coronary heart disease (CHD) severity, and major cardiovascular events (MACEs) in patients with type 2 diabetes mellitus (T2DM).MethodsA total of 2984 T2DM patients underwent selective coronary angiography, and their serum PCSK9 levels were measured using enzyme-linked immunosorbent assay. Correlation and logistic regression analyses were performed to investigate the association between PCSK9 expression and CHD severity. This study used Cox regression analysis to assess the association between circulating PCSK9 levels and the risk of MACEs.ResultsCirculating PCSK9 levels were significantly higher in the CHD group than in the non-CHD group [554.62 (265.11) ng/mL vs. 496.86 (129.05) ng/mL, p < 0.001]. Circulating PCSK9 levels positively correlated with CHD severity (diseased vessels: r = 0.35, p < 0.001; Gensini score: r = 0.46, p < 0.001). Elevated PCSK9 levels are an independent risk factor for CHD risk and severity (CHD group vs. non–CHD group: OR = 2.829, 95% CI: 1.771–4.520, p < 0.001; three vessel disease group vs. one vessel disease group: OR = 4.800, 95% CI: 2.387–9.652, p < 0.001; high GS group vs. low GS group: OR = 5.534, 95% CI: 2.733–11.208, p < 0.001). Through a six-year follow-up and multivariate Cox regression analysis, elevated circulating PCSK9 levels were found to be independently associated with MACEs in all participants (HR: 3.416, 5% CI: 2.485–4.697, p < 0.001; adjusted HR: 2.780, 95% CI: 1.930–4.004, p < 0.001).ConclusionsSerum PCSK9 levels were positively correlated with multi-vessel CHD and Gensini score. Elevated circulating PCSK9 levels are an independent risk factor for CHD and increased incidence of MACEs in T2DM.

The Immunogenic Potential of PCSK9 Peptide Vaccine in Mice.

To evaluate the immunogenic potential of the carrier-free peptide-based anti-PCSK9 (proprotein convertase subtilisin/kexin 9) vaccine in albino mice. The immunogenic pcsk9 peptide and 0.4% alum adjuvant were mixed thoroughly at a 1:1 ratio and used as a vaccine formulation. To assess the humoral immune response, animals' blood was sampled two weeks after the last immunization. The ELISA method was employed to measure serum anti-PCSK9 antibody titers, PCSK9 concentrations, and PCSK9/LDLR interaction. ELISA analysis showed significant induction of IgG antibody titers by PCSK9 peptide vaccine in vaccinated mice sera compared to the control mice (in male and female mice were 12000±586 and 11566±642, respectively, p<0.001). Mechanistic analyses showed a significant reduction in serum PCSK9 concentrations by vaccine-induced antibodies in vaccine groups compared to the control groups (in male mice by 29±5 ng/mL (22.4%), p<0.001 and female mice by 26±5 ng / mL (21.0%), p<0.001). Serum concentrations of PCSK9 in control and vaccine groups were 131±8.6 ng / mL and 102±8.1 ng/ml in male mice and 124±6 ng/ml and 98±10 ng/ml in female mice, respectively. Moreover, vaccine-induced antibodies inhibited the PCSK9-LDLR interaction in male and female groups by 34% and 26%, respectively. No significant difference was detected between the male and female groups in all tests (p>0.05). According to our results, the PCSK9 peptide vaccine provoked the humoral immune system in albino mice to produce functional antibodies that inhibit plasma PCSK9. These effects were seen in both genders without any significant difference.

Detrimental effects of PCSK9 loss-of-function in the pediatric host response to sepsis are mediated through independent influence on Angiopoietin-1

BackgroundSepsis is associated with significant mortality. Yet, there are no efficacious therapies beyond antibiotics. PCSK9 loss-of-function (LOF) and inhibition, through enhanced low-density lipoprotein receptor (LDLR) mediated endotoxin clearance, holds promise as a potential therapeutic approach among adults. In contrast, we have previously demonstrated higher mortality in the juvenile host. Given the potential pleiotropic effects of PCSK9 on the endothelium, beyond canonical effects on serum lipoproteins, both of which may influence sepsis outcomes, we sought to test the influence of PCSK9 LOF genotype on endothelial dysfunction.MethodsSecondary analyses of a prospective observational cohort of pediatric septic shock. Genetic variants of PCSK9 and LDLR genes, serum PCSK9, and lipoprotein concentrations were determined previously. Endothelial dysfunction markers were measured in day 1 serum. We conducted multivariable linear regression to test the influence of PCSK9 LOF genotype on endothelial markers, adjusted for age, complicated course, and low- and high-density lipoproteins (LDL and HDL). Causal mediation analyses to test impact of select endothelial markers on the association between PCSK9 LOF genotype and mortality. Juvenile Pcsk9 null and wildtype mice were subject to cecal slurry sepsis and endothelial markers were quantified.ResultsA total of 474 patients were included. PCSK9 LOF was associated with several markers of endothelial dysfunction, with strengthening of associations after exclusion of those homozygous for the rs688 LDLR variant that renders it insensitive to PCSK9. Serum PCSK9 was not correlated with endothelial dysfunction. PCSK9 LOF influenced concentrations of Angiopoietin-1 (Angpt-1) upon adjusting for potential confounders including lipoprotein concentrations, with false discovery adjusted p value of 0.042 and 0.013 for models that included LDL and HDL, respectively. Causal mediation analysis demonstrated that the effect of PCSK9 LOF on mortality was mediated by Angpt-1 (p = 0.0008). Murine data corroborated these results with lower Angpt-1 and higher soluble thrombomodulin among knockout mice with sepsis relative to the wildtype.ConclusionsWe present genetic and biomarker association data that suggest a potential direct role of the PCSK9-LDLR pathway on Angpt-1 in the developing host with septic shock and warrant external validation. Further, mechanistic studies on the role of PCSK9-LDLR pathway on vascular homeostasis may lead to the development of pediatric-specific sepsis therapies.Graphical abstract

Gene Expression and Methylation Levels of PCSK9 Gene in Iraqi Patients with Coronary Artery Disease

The expression of the Proprotein Convertase Subtilisin/Kexin Type 9 gene (PCSK9) is inextricably related to lipid levels and a risk of atherosclerotic coronary artery disease (ASCAD). The present study aims to measure the quantity of PCSK9 gene expression and the effect of methylation on its expression level taking part in the pathogenesis of acute coronary artery disorder. A current study included 150 subjects from the Iraqi population, 100 ASCAD patients and 50 healthy controls. The concentration of PCSK9 in each serum sample was determined by the ELISA technique, the expression levels of the PCSK9 gene in whole blood were estimated by RT-qPCR – Quantitative Reverse Transcription PCR method, and DNA methylation level detection in PCSK9 gene by using High Resolution Melting Analysis (HRM) technique. The expression level of the PCSK9 gene was 6.06 ± 1.84 in ASCAD patients group compared with a control group 1.00 ±0.19. That indicates the up-regulation of the PCSK9 gene in patient with atherosclerosis. In addition, the PCSK9 concentration was higher in patient group in comparison to the healthy controls group P= &lt; 0.01. However, PCSK9 methylation levels, a highly significant distinction between the two study groups p=0.002. In conclusion, serum PCSK9 levels are associated with coronary artery lesions as shown in receiver operating characteristic (ROC) analysis. Suggests that, serum PCSK9 might be a good indicator of coronary artery disease development.

The diagnostic value of echocardiography combined with serum Hcy and PCSK9 in coronary heart disease

ABSTRACT Coronary heart disease (CHD) is one of the most common cardiovascular diseases. This study analyzed the diagnostic value of echocardiography combined with serum homocysteine (Hcy) and proprotein convertase subtilisin 9 (PCSK9) in CHD. Here, 108 CHD patients were selected as the study group. Additionally, 108 patients with suspected CHD excluded by coronary angiography were selected as the control group. Serum Hcy and PCSK9 levels were detected by circulating enzymatic and biochemical analysis assay. The contrast echocardiography showed that the contrast agent filling velocity (β) and the maximum number of microbubbles (A) in the study group were declined compared to the control group. Serum Hcy and PCSK9 levels in the study group were higher than those in the control group. In addition, β, A, Hcy and PCSK9 are important influencing factors of CHD. The coronary artery disease branch numbers and the degree of stenosis in CHD patients were negatively correlated with β and A values, and positively correlated with serum Hcy and PCSK9 levels. β, A combined with serum Hcy and PCSK9 levels have diagnostic value for CHD, and are significantly correlated with the severity of CHD.

PCSK9 involves in the high-fat diet-induced abnormal testicular function of male mice.

Impaired spermatogenesis resulting from disturbed cholesterol metabolism due to intake of high-fat diet (HFD) has been widely recognized, however, the role of preprotein invertase subtilin 9 (PCSK9), which is a negative regulator of cholesterol metabolism, has never been reported. This study aims to reveal the role of PCSK9 on spermatogenesis induced by HFD in mice. Long-term consumption of a high-fat diet (HFD) is an important factor that leads to impaired spermatogenesis exhibiting poor sperm quantity and quality. However, the mechanism of this is yet to be elucidated. Disrupted cholesterol homeostasis is one of many crucial pathological factors which could contribute to impaired spermatogenesis. As a negative regulator of cholesterol metabolism, preprotein invertase subtilin 9 (PCSK9) mediates low density lipoprotein receptor (LDLR) degradation to the lysosome, thereby reducing the expression of LDLR on the cell membrane and increasing serum low-density lipoprotein cholesterol level, resulting in lipid metabolism disorders. Here, we aim to study whether PCSK9 is a pathological factor for impaired spermatogenesis induced by HFD and the underlying mechanism. To meet the purpose of our study, we utilized wild-type C57BL/6 male mice and PCSK9 knockout mice with same background as experimental subjects and alirocumab, a PCSK9 inhibitor, was used for treatment. Results indicated that HFD induced higher PCSK9 expression in serum, liver, and testes, and serum PCSK9 is negatively correlated with spermatogenesis, while both PCSK9 inhibitor treatment and PCSK9 knockout methodologies ameliorated impaired lipid metabolism and spermatogenesis in mice fed a HFD. This could be due to the overexpression of PCSK9 induced by HFD leading to dyslipidemia, resulting in testicular lipotoxicity, thus activating the Bcl-2-Bax-Caspase3 apoptosis signaling pathway in testes, particularly in Leydig cells. Our study demonstrates that PCSK9 is an important pathological factor in the dysfunction of spermatogenesis in mice induced by HFD. This finding could provide innovative ideas for the diagnosis and treatment of male infertility.

Abstract 15273: Impact of Lipid Lowering Therapy on Progenitor Endothelial Cell Phenotype and Function in Coronary Artery Disease

Introduction: Early revascularization of infarcted zone is crucial to improve patient′s outcomes. Despite their primary actions, major lipid-lowering drugs may also reduce cardiovascular events by pleiotropic mechanisms. Hypothesis: Conventional (statin) and novel lipid-lowering therapies [(PSCK9-inhibitor (PCSK9i) and eicosatetraenoic acid (EPA)] can improve endothelial progenitor cells (EPC) phenotype and favor neovascularization. Methods: EPC were isolated from peripheral blood of patients with acute coronary syndrome (n=62) who were statin-naïve (n=34) or chronic statin treatment (n=28) at time of inclusion. A set of statin-naïve EPC and mature endothelial cells (HUVEC) were treated in vitro with statin (20nM), PCSKi (1μM) and EPA (3μM). EPC phenotype was determined as EPC colony number and time of appearance, proliferative rate, and ability to form capillary-like structures. Serum PCSK9 and markers of angiogenesis and endothelial function were determined by multiplexed protein assay. Results: Frequency of EPC appearance was higher in samples of statin group (78%) compared to naïve group (66%) (Fig A). They also appeared much earlier (Fig B), in greater quantity (4.7 vs 2.1 colony, p = 0.0009) and had higher proliferative rate than EPCs of naïve group. Besides they had increased potential of tube formation and shaped more branched vessel-like structures (Fig C). No differences in serum levels of biomarkers were found. In vitro treatment of EPC with statin, PCSK9i and EPA did not modify colonies behavior (Fig D). PCSK9 levels showed negative correlation with EPC proliferation (r -0.4150, p=0.004), and PCSK9i treatment increased angiogenic potential of HUVEC (Fig E). Conclusions: Statin and PCSK9i have pleiotropic effects on EPC, boosting their potential to form endothelial cells colonies and to induce angiogenesis. These effects could contribute to the progression and outcomes of coronary artery disease. Funding: ISCIII - FEDER-ERDF (PI19/00264)

ODP557 PCSK9 Level Correlated with Disease Progression in Patient with Non-small Cell Lung Cancer

Abstract Background Hyperlipidemia may be present in cancer patients despite their debilitated general condition. It has also been reported that several types of cancer cells produce PCSK9, and that in advanced non-small cell lung cancer, chemotherapy with immune checkpoint inhibitors is effective in cases with low PCSK9 levels. We administered PCSK9 inhibitor to patients with advanced non-small cell lung cancer with hyperlipidemia according to the indications for hyperlipidemia. Chemotherapy with immune checkpoint inhibitors was performed, the patient's condition and the level of PSCK9 were observed. CASE: A woman in her 60s underwent surgery for lung cancer in 2017. The pathological diagnosis of squamous cell carcinoma of the left lower bronchus and the postoperative diagnosis of pT2bN2M0 stage IIIA were confirmed. Adjuvant chemotherapy with CDDP and VNR was started after surgery for her, but in 2018, recurrence of pulmonary lymph nodes was confirmed by intrabronchial induced transbronchial needle aspiration. The immunostaining result for PD-L1 was 50%. The first-line chemotherapy, the immune checkpoint inhibitor pembrolizumab, was started but was ineffective. She had high serum LDL levels of 174 mg/dL and high serum PCSK9 levels of 471 ng /mL (Norm: 47.4-225.2 ng/mL) despite exhaustion during her lung cancer treatment process. Her plasma PCSK9 levels were correlated with lung cancer status, suggesting PCSK9 production in lung cancer cells. Ezetimibe and statins were also discontinued because they were reported to increase serum PCSK9. The PCSC9K inhibitor evolocumab was administered at 140 mg every 2 weeks. Three months after the start of evolocumab, her serum PCSK9 level decreased slightly to 441 ng /mL. The immune checkpoint inhibitor nivolumab was then started, but the chemotherapy was ineffective, and her general condition worsened, correlating with her serum PCSK9 levels rising to 1635 ng/mL, and she died in 2021. Discussion and Clinical Lessons This is the first report on the clinical course of non-small cell lung cancer and the course of serum PCSK9 and the administration of PCSK9 inhibitors. Although PSCK9 is thought to be involved in cancer progression, the use of PCSK9 inhibitors at the same doses and doses as in the treatment of conventional hyperlipidemia failed to control cancer progression. The anti-PCSK9 vaccine has been experimentally shown to be effective against tumor progression. It is expected that PCSK9 inhibition by various methods will bring good results for cancer patients. Further research is needed to elucidate the exact mechanism by which PCSK9 regulates cancer cell proliferation and apoptosis. PCSK9 inhibitors, initially developed to treat hypercholesterolemia, may have significant involvement in the treatment of various cancers and their metastases. Therefore, collaboration between endocrinologists and oncologists is important in the laboratory and clinical practice. Presentation: No date and time listed

The immunogenic potential of PCSK9 peptide vaccine in mice

Abstract Background Proprotein convertase subtilisin/Kexin 9 (PCSK9) is a serum protein expressed by liver cells that regulates plasma low-density lipoprotein cholesterol (LDL-C) levels by modulating LDL receptors (LDLR). Recent studies of our group indicated the efficient immunogenicity of the nanoliposomal PCSK9 peptide vaccine in different animal models. Purpose Herein, we sought to evaluate the immunogenic potential of the non-liposomal form of the vaccine due to its simplicity of processing and lower production cost. Methods PCSK9 immunogenic peptide was prepared by linking a short PCSK9 peptide to a tetanus peptide as B and T cell epitopes, respectively. The immunogenic peptide and 0.4% alum adjuvant, in a 1:1 ratio, were mixed thoroughly and homogeneously prior to administration as vaccine formulation. In order to assess the humoral immune response, mice were inoculated subcutaneously four times at bi-week intervals. Immunized animals' blood was sampled two weeks after the last injection. The titers of the anti-PCSK9 antibodies in the serum of vaccinated mice were evaluated using the ELISA method. ELISA was also used to investigate the effects of anti-PCSK9 vaccination on PCSK9 serum concentrations and its interaction with LDL receptor (LDLR). Results ELISA analysis showed significant induction of IgG antibody titers by PCSK9 peptide vaccine in vaccinated mice compared to the control group (586±12000 and 642±11566 in male and female vaccinated mice, respectively, p&amp;lt;0.001). No significant difference was found between the male and female vaccine groups (p&amp;gt;0.05). Mechanistic analyses showed that vaccine-induced antibodies decreased serum PCSK9 concentration in vaccine groups compared to the control group (in male mice by 26±5 ng/mL, p&amp;lt;0.01 and female mice by 29±5 ng/mL, p&amp;lt;0.01). Serum concentrations of PCSK9 in control and vaccine groups were found to be 131±8.6 ng/mL and 102±8.1 ng/ml in male mice, and 124±6 ng/ml and 98±10 ng/ml in female mice, respectively. Moreover, vaccine-induced antibodies inhibited the PCSK9/LDLR interaction in the presence of serum from vaccinated mice compared to control mice serum (in male and female groups by 34% and 26%, respectively). The PCSK9 peptide vaccine significantly reduced LDL-C in immunized mice in both male (16.93±2.5 mg/dL, p=0.001) and female groups (22±2.7 mg/dL, p=0.001) compared with the control group. Conclusions According to our results, the PCSK9 peptide vaccine stimulates the humoral immune response in albino mice to produce active antibodies that inhibit plasma PCSK9 resulting in plasma LDL-C reduction. Funding Acknowledgement Type of funding sources: None.

Does pathological type of primary nephrotic syndrome affect serum concentrations of proprotein convertase subtilisin/kexin type 9?

BackgroundDyslipidemia is a common finding in primary nephrotic syndrome (PNS). Serum PCSK9 level is also increased in PNS and is the main cause of dyslipidemia in such patients. There is a paucity of data on the relation between dyslipidemia and pathological types of PNS. We hypothesized that severity of dyslipidemia varies across different types of PNS, and this variation is due to differences in serum PCSK9 levels.MethodsFifty patients recently diagnosed with PNS were included in this cross-sectional study. Serum PCSK9, albumin, creatinine, total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), very low-density lipoprotein cholesterol (VLDL-C), triglycerides (TG), and 24-h urine protein were measured. Comparison of patients’ characteristics between pathological types of PNS and correlations between serum PCSK9 and other parameters were assessed.ResultsSerum PCSK9 levels were significantly higher in PNS patients compared with healthy individuals (314.58 ± 73.83 vs 253.42 ± 36.66 ng/ml, p < 0.001). No differences found between PNS types regarding serum levels of PCSK9 (p = 0.571), TC (p = 0.806), LDL-C (p = 0.950), HDL-C (p = 0.844), VLDL-C (p = 0.472), and TG (p = 0.969). Serum PCSK9 levels correlated significantly with TC (p < 0.001), LDL-C (p < 0.001), HDL-C (p = 0.003), VLDL-C (p = 0.008), TG (p = 0.005), 24-h urine protein (p = 0.005), and male sex (p = 0.014).ConclusionThe pathological type of PNS does not affect serum levels of PCSK9 and components of lipid profile.

Evaluation of serum and salivary PCSK9 and IL6 and its association with periodontal inflammation and atherosclerotic cardiovascular diseases.

and Purpose: The aim of this study was to assess, compare and correlate the ability of PCSK9 and IL6 as potential common serum and salivary biomarkers for stage III/IV periodontitis and the risk of developing atherosclerotic cardiovascular diseases. 76 patients were alloted into 4 groups consisting of subjects with clinically healthy periodontium and stage III/IV periodontitis with and without ACVD. Clinical parameters - PD, CAL, number of teeth, PI, mSBI, TC, Tg, HDL and LDL were recorded. Serum and saliva samples were obtained and subjected to ELISA for quantifying the biomarker levels. The level of these biomarkers was found to be the lowest (IL6 Serum: 21.92±14.54, IL6 Saliva: 12.34±7.72, PCSK9 Serum: 178.82±35.07, PCSK9 Saliva: 80.82±25.43) in group I and highest in group IV (IL6 Serum: 73.4±24.86, IL6 Saliva: 37.66±15.77, PCSK9 Serum: 346.54±45.11, PCSK9 Saliva: 157.72±43.28). The clinical parameters PI, mSBI, PPD, CAL, total number of teeth, TC, HDL and LDL showed a significant correlation with biomarkers PCSK9 and IL6. This study features the elevation of serum and salivary PCSK9 and IL6 in periodontitis and ACVD. A triad of serum and salivary PCSK9 and IL6 along with the clinical markers of periodontal disease can relatively predict the future risk of developing ACVD.

Plasma Proprotein Convertase Subtilisin/kexin Type 9 (PCSK9) in the Acute Respiratory Distress Syndrome.

BackgroundProprotein convertase subtilisin/kexin type 9 (PCSK9) is a serine protease that is a mediator of the immune response to sepsis. PCSK9 is also highly expressed in pneumocytes and pulmonary endothelial cells. We hypothesized that serum PCSK9 levels would be associated with death and ICU outcomes in patients with ARDS.MethodsUsing data and plasma samples from the NIH BioLINCC data repository, we assembled a cohort of 1,577 patients with the acute respiratory distress syndrome (ARDS) enrolled in two previously completed clinical trials, EDEN and SAILS. We measured PCSK9 levels in plasma within 24 h of intubation using commercially available ELISA kits (R&D Systems). We assessed the association of PCSK9 with mortality using Cox proportional hazard models. We also assessed clinical factors associated with PCSK9 level and the association of PCSK9 with the number of days free of mechanical ventilation and days free of ICU care.ResultsIn 1,577 ARDS patients, median age was 53 years (IQR 42–65 years) and median APACHE III score 91 (72–111) connoting moderate critical illness. PCSK9 levels were 339.3 ng/mL (IQR 248.0–481.0). In multivariable models, race, cause of ARDS, body mass index, pre-existing liver disease, body temperature, sodium, white blood cell count and platelet count were associated with PCSK9 level. Presence of sepsis, use of vasopressors and ventilator parameters were not associated with PCSK9 level. PCSK9 levels were not associated with in-hospital mortality (HR per IQR 0.96, 95% CI 0.84–1.08, P = 0.47). Higher PCSK9 levels were associated with fewer ICU and ventilator free days.ConclusionsPlasma PCSK9 is not associated with mortality in ARDS, however higher PCSK9 levels are associated with secondary outcomes of fewer ICU free and ventilator free days. Clinical factors associated with PCSK9 in ARDS are largely unmodifiable. Further research to define the mechanism of this association is warranted.

Time-averaged low-density lipoprotein cholesterol lowering with evolocumab: Pooled analysis of phase 2 trials

LDL-C is the pivotal risk factor for atherosclerotic cardiovascular disease, and the benefit from LDL-C lowering is proportional to the magnitude of reduction. Clinical trials demonstrate that evolocumab reduces LDL-C levels by approximately 60% when measured at the trough of drug effect, which may underestimate cumulative LDL-C reduction. We obtained a time-averaged estimate of LDL-C lowering that included both peaks and troughs. Pooled analysis of 5 phase 2 trials included patients with hypercholesterolemia who received placebo or evolocumab (140 mg every 2 weeks [Q2W] or 420 mg monthly [QM]). Percent changes from baseline LDL-C and free serum PCSK9 were averaged across weeks 9–12. In 372 patients, time-averaged percent reduction from baseline in LDL-C with evolocumab vs placebo was 67.6% (95% CI: 63.9–71.3) with Q2W dosing and 65.0% (95% CI: 60.7–69.3) with QM dosing. The time-averaged measure yielded LDL-C reductions for evolocumab that exceeded measurements at the end of dosing intervals and may provide a better estimate of cardiovascular benefit during long-term therapy.

MO244: PCSK9 Inhibitors are Efficient in Treatment of Resistant Hypercholesterolemia in Nephrotic Syndrome

Abstract BACKGROUND AND AIMS Hypercholesterolemia in nephrotic syndrome is commonly resistant to statin therapy and represents an important clinical challenge both for cardiovascular risk prevention and to eliminate lipotoxic effect on podocytes. Serum PCSK9 level is elevated in nephrotic syndrome and has a key pathophysiologic role in development and persistance of hyperlipidemia. Thus, we aimed to investigate the role of PCSK9 inhibitors in control of hypercholesterolemia in patients with neprotic syndrome. METHOD Consecutive patients with nephrotic syndrome and elevated atherogenic Lp (a) (n = 4) and patients with resistant secondary glomerulonephritis with nephrotic proteinuria (n = 2) were included. In all patients, in addition to full dose statin therapy, evolocumab 420 mg s.c. monthly was introduced. Lipid levels were measured after a median of 4 (range 2–9) months after starting evolocumab. RESULTS Nephrotic syndrome was caused by focal segmental glomerulosclerosis in four patients (66.7%) and membranous nepropathy was the cause in two patients (33.3%). Median age of included patients was 49.5 (range 24–70) years. Four patients with primary cause of glomerulonephritis received specific imunosupressive treatment. During follow-up, we observed descrease in glomerular filtration rate (eGFR median 58.5 interquartile range (IQR) [46.7–98.7] mL/min/1.73 m2 versus median 47 IQR [38.25–89.25] mL/min/1.73 m2; P = .046) and persistence of proteinuria [median 11 970 IQR (3862–13 765) mg/dU versus median 10 847 (5407–14 340) mg/dU; P = .753]. However, despite persistent proteinuria and decrease in eGFR, significant decrease in total cholesterol [median 9.10 IQR (6.92–10.87) mmol/L versus median 6.81 (5.4–8.45) mmol/L; P = .028] and LDL cholesterol [median 5.40 IQR (4.12–7.25) versus median 4.04 IQR (2.70–4.60); P = .046] was observed. There was no significant difference in Lp (a) and triglyceride levels. CONCLUSION PCSK9 inhibitors are a promising new therapeutic option for hyperlipidemia in patients with nephrotic syndrome. Further research, including a larger number of patients, is needed to correctly position PCSK9 inhibitors in treatment of hyperlipidemia in patients with nephrotic syndrome.

Relationship of sortilin and proprotein convertase subtilisin/kexin type 9 in blood serum with the severity of carotid and coronary atherosclerosis in hypertensive patients

Sortilin is an important molecular protein involved in lipid metabolism, atherosclerosis, and aortic valve calcification. Sortilin presumably regulates the PCSK9 signaling pathways.Aim. To study correlations of sortilin and PCSK9 with atherosclerosis development in hypertensive patients.Material and methods. The study included 161 patients aged 30 to 65 years. We performed collection of complaints and anamnesis, physical examination, blood biochemical test with the determination of total cholesterol, low-density lipoprotein cholesterol, triglycerides, blood glucose, serum creatinine with estimation of glomerular filtration rate. Serum PCSK9, sortilin and interleukins 8, 10 were determined by enzyme-linked immunosorbent assay. The following investigations were also performed: electrocardiography, echocardiography, extracranial artery ultrasound, coronary angiography.Results. Sortilin levels (b=2,37; odds ratio (OR), 10,74; 95% CI, 1,05-109,47, p=0,045), IL-8 (b=-2,42; OR, 9,74; 95% CI, 0,01-0,81, p=0,032), age (b=0,21; OR, 1,24; 95% CI, 1,12-1,37, p&lt;0,001) were identified as independent predictors of coronary atherosclerosis with a sensitivity of 87% and a specificity of 70%. PCSK9 (b=0,005; OR, 1,00; 95% CI, 1,00-1,01, p=0,038) and IL-8 (b= -0,33; OR, 0,72; 95% CI, 0,55-0,94, p=0,014) were identified as independent predictors of carotid atherosclerosis with a sensitivity of 75% and a specificity of 71%.Conclusion. In addition to non-invasive imaging, the determination of atherosclerosis biomarkers can make a significant contribution to the diagnosis and prediction of carotid and coronary atherosclerosis progression. It is noteworthy that not only PCSK9, but also sortilin can be a potential therapeutic target. Further large-scale studies are needed.

Prevention of Triglyceridemia by (Non-)Anticoagulant Heparin(oids) Does Not Preclude Transplant Vasculopathy and Glomerulosclerosis.

Background: In renal transplantation, chronic transplant dysfunction (CTD) is associated with increased PCSK9 and dyslipidemia. PCSK9 is an enzyme that increases plasma cholesterol levels by downregulating LDLR expression. We recently showed increased PCSK9–syndecan-1 interaction in conditions of proteinuria and renal function loss. Treatment with heparin(oids) might be a therapeutic option to improve dyslipidemia and CTD. We investigated the effects of (non-)anticoagulant heparin(oids) on serum lipids, syndecan-1 and PCSK9 levels, and CTD development. Methods: Kidney allotransplantation was performed from female Dark Agouti to male Wistar Furth recipients. Transplanted rats received daily subcutaneous injections of saline, unfractionated heparin, and RO-heparin or NAc-heparin (2 mg heparin(oid)/kg BW) until sacrifice after 9 weeks of treatment. Results: Saline-treated recipients developed hypertension, proteinuria, and loss of creatinine clearance (all p < 0.05 compared to baseline), along with glomerulosclerosis and arterial neo-intima formation. Saline-treated recipients showed significant increase in plasma triglycerides (p < 0.05), borderline increase in non-HDLc/HDLc (p = 0.051), and ∼10-fold increase in serum syndecan-1 (p < 0.05), without significant increase in serum PCSK9 at 8 weeks compared to baseline. Heparin and non-anticoagulant RO-heparin administration in transplanted rats completely prevented an increase in triglycerides compared to saline-treated recipients at 8 weeks (both p < 0.05). Heparin(oids) treatment did not influence serum total cholesterol (TC), plasma syndecan-1 and PCSK9 levels, creatinine clearance, proteinuria, glomerulosclerosis, and arterial neo-intima formation, 8 weeks after transplantation. Combining all groups, increased syndecan-1 shedding was associated with TC (r = 0.5; p = 0.03) and glomerulosclerosis (r = 0.53; p = 0.021), whereas the non-HDLc/HDLc ratio was associated with the neo-intimal score in the transplanted kidneys (r = 0.65; p < 0.001). Conclusion: Prevention of triglyceridemia by (non-)anticoagulant heparin(oids) neither influenced PCSK9/syndecan-1 nor precluded CTD, which however did associate with the shedding of lipoprotein clearance receptor syndecan-1 and the unfavorable cholesterol profile.