Addition Of Simvastatin Research Articles

Long-term hemodynamic response of carvedilol and simvastatin in compensated versus decompensated chronic liver disease with portal hypertension

Background: A combination of carvedilol and Simvastatin have recognized their role as rescue therapy for carvedilol non-responders in chronic liver disease (CLD)-associated portal hypertension. However, there are scarce data regarding its role in specific subclasses, and stratification of response according to the computed tomographic perfusion (CTP) score has never been seen before. Aims and Objective: (1) To compare hemodynamic response and side effects of a combination therapy in decompensated and compensated patients. (2) Stratify the effect of combination therapy according to CTP score. Materials and Methods: In 102 consecutive patients of CLD with esophageal varices, the hepatic venous pressure gradient was measured at baseline and after 3 months of dose optimization of carvedilol. Simvastatin was added to non-responders and hemodynamics repeated at 1 month of dual therapy. The response of compensated CLD was compared with decompensated patients and was stratified as per the CTP on follow-up. Results: Overall, out of 43 compensated patients, 21 responded acutely and response increased to 29 (67.44%) at 3 months. While 31 of 59 (52.54%) decompensated patients responded acutely but dose escalation did not increase response significantly after 3 months. The addition of Simvastatin did increase the response, although side effects were more in decompensated group. The addition of Simvastatin also decreased decompensation in the high-risk compensated group and maintained their MELD.Na. Conclusion: Response in compensated CLD patients was more than in decompensated patients; however, it was statistically insignificant. Attrition was more in patients with CTP >10 due to drug intolerance, side effects, or deaths.

Simvastatin Overcomes Resistance to Tyrosine Kinase Inhibitors in Patient-derived, Oncogene-driven Lung Adenocarcinoma Models.

There is an unmet clinical need to develop novel strategies to overcome resistance to tyrosine kinase inhibitors (TKI) in patients with oncogene-driven lung adenocarcinoma (LUAD). The objective of this study was to determine whether simvastatin could overcome TKI resistance using the in vitro and in vivo LUAD models. Human LUAD cell lines, tumor cells, and patient-derived xenograft (PDX) models from TKI-resistant LUAD were treated with simvastatin, either alone or in combination with a matched TKI. Tumor growth inhibition was measured by the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay and expression of molecular targets was assessed by immunoblots. Tumors were assessed by histopathology, IHC stain, immunoblots, and RNA sequencing. We found that simvastatin had a potent antitumor effect in tested LUAD cell lines and PDX tumors, regardless of tumor genotypes. Simvastatin and TKI combination did not have antagonistic cytotoxicity in these LUAD models. In an osimertinib-resistant LUAD PDX model, simvastatin and osimertinib combination resulted in a greater reduction in tumor volume than simvastatin alone (P < 0.001). Immunoblots and IHC stain also confirmed that simvastatin inhibited TKI targets. In addition to inhibiting 3-hydroxy-3-methyl-glutaryl-CoA (HMG-CoA) reductase, RNA sequencing and Western blots identified the proliferation, migration, and invasion-related genes (such as PI3K/Akt/mTOR, YAP/TAZ, focal adhesion, extracellular matrix receptor), proteasome-related genes, and integrin (α3β1, αvβ3) signaling pathways as the significantly downregulated targets in these PDX tumors treated with simvastatin and a TKI. The addition of simvastatin is a safe approach to overcome acquired resistance to TKIs in several oncogene-driven LUAD models, which deserve further investigation.

Effect of Simvastatin on Hepatic Venous Portal Gradient for Maintenance of Lipid Profile of Liver Cirrhosis patients

Background: Liver cirrhosis has an essential influence on the lipid profile. Statins have well-known beneficial cardiovascular effects reducing cardiovascular events and mortality. Aim: To compare the change in hepatic venous portal gradient with or without simvastatin for maintenance of lipid profile of liver cirrhosis patients Methods: This Randomized controlled trial was done at Department of Medicine, South medical ward, Mayo hospital, Lahore from October 2021 to March 2022.patients were randomly divided in two groups. In group A, patients were given simvastatin with standard treatment. In group B, patients were given standard treatment only. Before and after 1 month of treatment, lipid profile and HVPG level were assessed again. The change in HVPG and lipid profile was calculated. Data was entered and analyzed in SPSS version 25. Results: The mean age of patients received simvastatin was 55.26 ± 13.24 years and mean age of patients in control was 53.74 ± 16.88 years. In simvastatin group, there were 82 (54.7%) males and 68(45.3%) females. In control group, there were 93 (62.0%) males and 57(38%) females. With simvastatin, the mean HVPG level was reduced from 20.11± 6.21mg/dl to 15.42±4.71 mg/dl after a month (mean change = 4.69 ± 1.50 mg/dl). While in control group, mean HVPG level was reduced from 19.28±7.61 mg/dl to 17.82± 6.52 mg/dl after a month (mean change = 1.46±1.09 mg/dl). The effect size was 23.3% with simvastatin while 7.6% without simvastatin for reduction of HVPG in cirrhotic patients. The difference was observed to be significant (p&lt;0.05). Conclusion: Thus, addition of simvastatin is effective in controlling lipid profile of patients with liver cirrhosis. It also has beneficial role in improving condition of liver cirrhosis patients. Keywords: hepatic venous portal gradient, simvastatin, lipid profile, liver cirrhosis, LDL, HDL, total cholesterol, triglycerides

Simvastatin and fluvastatin attenuate trauma-induced cell death and catabolism in human cartilage.

Joint injuries are known to induce pathomechanisms that might lead to posttraumatic osteoarthritis (PTOA). In this regard, statins with their pleiotropic effects could represent potential therapeutic agents in preventing the development of PTOA. Therefore, we investigated the effects of simvastatin and fluvastatin in a drop-tower-based human ex vivo cartilage trauma model. After 7 days, a mechanical impact (0.59 J) resulted in a decrease of the cell viability and increased expression of catabolic enzymes in cartilage explants. Simvastatin and fluvastatin treatment of impacted cartilage demonstrated cell protective effects in a concentration dependent manner. Moreover, statin therapy exhibited chondroprotective effects as demonstrated by attenuated expression of MMP-2 and MMP-13 as well as subsequent breakdown of collagen type II (after impact). Further analysis indicated antioxidative properties of the statins by upregulating the gene expression of SOD2 and suppression that of NOX2 and NOX4. Despite its protective effects, simvastatin impaired the biosynthesis of collagen type II, which was confirmed during chondrogenic redifferentiation of high passage chondrocytes. However, while long-term administration of statins for 4 weeks impaired chondrogenic redifferentiation, addition of simvastatin at low concentrations for 1 week exhibited a slightly promoting effect. In conclusion, our data imply that simvastatin and fluvastatin are suitable in terms of initial harm reduction after cartilage trauma.

Modulation of the plasma lipidomic profile with simvastatin in metastatic castration-resistant prostate cancer (mCRPC).

154 Background: Elevated circulating sphingolipids are associated with poorer outcomes across the natural history of prostate cancer (PC), including metastatic relapse in localised PC, earlier androgen deprivation failure in metastatic hormone-sensitive PC, and shorter overall survival (OS) in mCRPC. We have derived and validated a poor prognostic 3-lipid signature (3LS) [consisting of ceramide Cer(d18:1/24:1), sphingomyelin SM(d18:2/16:0) and phosphatidylcholine PC(16:0/16:0)], which was independently associated with shorter radiographic progression-free survival (rPFS) and OS in men with mCRPC commencing taxanes or androgen receptor signaling inhibitors (ARSI). Statins significantly reduce plasma levels of ceramides, sphingomyelin and cholesterol in cardiovascular disease. We hypothesised that this therapy could change the poor prognostic lipid profile of patients with mCRPC. This study assessed whether the addition of simvastatin to standard treatment for mCRPC modulates the circulating lipidomic profile. Methods: This investigator-initiated, multi-centre, single arm, pilot study enrolled men with mCRPC commencing taxanes or ARSI for disease progression, who were not on a lipid-lowering agent. Men were treated with simvastatin 40mg orally once daily for 12 weeks, commencing on day 1 of treatment for mCRPC. Plasma was taken at baseline and after 12 weeks of simvastatin, and underwent lipidomic profiling of ̃800 lipids. Differences in lipid levels between baseline and post-simvastatin samples and between those with and without the 3LS were assessed using t-tests. Results: 27 men (74% on taxanes, 26% on ARSI) were recruited between May 2018 to March 2021. 46% of the men had the poor prognostic 3LS at baseline, of whom 45% lost the 3LS after simvastatin. Comparison between all paired baseline and post-simvastatin samples showed significant reduction (p &lt; 0.05) in free cholesterol, cholesteryl esters and some sphingolipids (sphingomyelins, hexosylceramides) with simvastatin treatment. Baseline profiles with the 3LS displayed significantly higher levels (p &lt; 0.05) of ceramides, hexosylceramides and sphingomyelins, relative to baseline profiles without the 3LS. Men who lost the 3LS after treatment (n = 5) demonstrated significant reductions in ceramides (23-45%, p≤0.046), hexosylceramides (27-52%, p≤0.049) and sphingomyelins (28-44%, p≤0.047). These changes were not seen in men with persistent 3LS after treatment (n = 6). Conclusions: Simvastatin in addition to standard treatment for mCRPC can modulate the circulating lipidomic profile and eliminate the presence of a poor prognostic 3LS in 45% of participants with the 3LS. Further prospective randomised control studies are required to determine if modulation of the 3LS by simvastatin can improve clinical outcomes. Clinical trial information: ACTRN12617000965303.

Analysis of Efficacy, Safety, and Prognostic Factors of mFOLFOX6 Regimen Combined with Cetuximab and Simvastatin in the Treatment of K-RAS Mutant Colorectal Cancer.

Colorectal cancer (CRC) is one of the most common malignant tumors with high morbidity and mortality. The early symptoms are latent, and most patients are in the middle or late stage when they are diagnosed. The best opportunity for surgery has been lost, and surgical resection has failed to achieve good results. In clinical practice, targeted therapy or chemotherapy is usually the main treatment. The mFOLFOX6 regimen is a standardized regimen for the treatment of advanced CRC. The main drugs in this regimen are oxaliplatin and 5-fluorouracil (5-FU). Patients with advanced CRC combined with standard chemotherapy regimens can achieve a higher resection rate of liver metastases in unresectable patients, which can achieve significant survival improvement. Therefore, in this study, oxaliplatin + calcium folinate + 5-Fu + mFOLFOX6 regimen was combined with cetuximab and simvastatin to treat CRC patients, and the clinical efficacy and prognosis were analyzed, as well as the prognostic factors. The results showed that the addition of simvastatin on the basis of conventional mFOLFOX6 regimen combined with cetuximab chemotherapy could effectively improve the efficacy, reduce the total incidence of adverse reactions, improve the overall survival rate, and prolong the overall survival time of patients. Pathological grade and peritoneal metastasis were the factors affecting the mean survival time of CRC patients.

Hydroxyapatite-Based Magnetic Bionanocomposite as Pharmaceuticals Carriers in Chitosan Scaffolds

Hydroxyapatite (HA) is a bioceramic very similar to the mineral component of bones and teeth. It is well established that osteoblasts grow better onto HA-coated metals than on metals alone. Herein, the preparation of a new system consisting of magnetite (Fe3O4) and HA functionalized with oleic acid and simvastatin (SIMV), and incorporated in chitosan (CHI) scaffolds, was undertaken. HA was synthesized by the hydrothermal method, while Fe3O4 was synthesized by co-precipitation. The polymer matrix was obtained using a 2% CHI solution, and allowed to stir for 2 h. The final material was freeze-dried to produce scaffolds. The magnetic properties remained unchanged after the formation of the composite, as well as after the preparation of the scaffolds, maintaining the superparamagnetism. CHI scaffolds were analyzed by scanning electronic spectroscopy (SEM) and showed a high porosity, with very evident cavities, which provides the functionality of bone growth support during the remineralization process in possible regions affected by bone tissue losses. The synthesized composite showed an average particle size between 15 and 23 nm for particles (HA and Fe3O4). The scaffolds showed considerable porosity, which is important for the performance of various functions of the tissue structure. Moreover, the addition of simvastatin in the system can promote bone formation.

Modulation of plasma lipidomic signature in metastatic castration-resistant prostate cancer (mCRPC).

TPS331 Background: Altered lipid metabolism and its impact on prostate cancer (PC) is increasingly recognised, in light of the association between obesity and worse PC outcomes. Our exploratory study was the first to identify baseline plasma lipidomic profiles in men with mCRPC commencing docetaxel that were associated with survival. A prognostic three-lipid signature was derived, consisting of ceramide, sphingomyelin and phosphatidylcholine (HR 4.8, 95% CI 2.06-11.1, p = 0.0003). This signature was independently prognostic when modelled with clinicopathological factors and metabolic characteristics. A key question is whether therapeutic modulation of a patient’s lipid profile is possible. Statins significantly reduce the plasma levels of ceramides, sphingomyelin and cholesterol in cardiovascular disease, suggesting that this therapy could change the poor prognostic lipid profile of mCRPC patients. This trial assesses whether addition of simvastatin to docetaxel for mCRPC can reverse the poor prognostic lipid signature with the aim of developing a precision medicine strategy for metabolic targeting. Methods: This investigator-initiated, multi-centre, open-label, single arm, pilot study enrols patients with mCRPC commencing docetaxel for disease progression, not already receiving a lipid-lowering agent. Patients are treated with simvastatin 40mg orally once daily for 12 weeks, commencing on day 1 of the first cycle of docetaxel. Blood is taken at baseline and after 12 weeks of simvastatin and the plasma lipidomic profile is determined using liquid chromatography and electrospray ionisation-tandem mass spectrometry. The lipidomic profile is classified as either good or poor prognostic as per our three-lipid signature model derived by logistic regression. The primary objective is to assess the rates of conversion from a poor prognostic lipid signature to good prognostic after simvastatin. A sample size of 60 men provides over 90% power, with a 1-sided type 1 error of 10%, to detect conversion to the good prognostic signature in 50% of patients, assuming 25% of patients have the poor prognostic signature at baseline as previously detected. To date, 6 patients have been enrolled to the trial. Clinical trial information: ACTRN12617000965303.

The addition of simvastatin administration to cold storage solution of explanted whole liver grafts for facing ischemia/reperfusion injury in an area with a low rate of deceased donation: a monocentric randomized controlled double-blinded phase 2 study

BackgroundLiver transplantation is the best treatment for end-stage liver disease. The interruption of the blood supply to the donor liver during cold storage damages the liver, affecting how well the liver will function after transplant. The drug Simvastatin may help to protect donor livers against this damage and improve outcomes for transplant recipients. The aim of this study is to evaluate the benefits of treating the donor liver with Simvastatin compared with the standard transplant procedure.Patient and methodsWe propose a prospective, double-blinded, randomized phase 2 study of 2 parallel groups of eligible adult patients. We will compare 3-month, 6-month, and 12-month graft survival after LT, in order to identify a significant relation between the two homogenous groups of LT patients. The two groups only differ by the Simvastatin or placebo administration regimen while following the same procedure, with identical surgical instruments, and medical and nursing skilled staff. To reach these goals, we determined that we needed to recruit 106 patients. This sample size achieves 90% power to detect a difference of 14.6% between the two groups survival using a one-sided binomial test.DiscussionThis trial is designed to confirm the effectiveness of Simvastatin to protect healthy and steatotic livers undergoing cold storage and warm reperfusion before transplantation and to evaluate if the addition of Simvastatin translates into improved graft outcomes.Trial registrationISRCTN27083228.

Trimetazidine restores the positive adaptation to exercise training by mitigating statin-induced skeletal muscle injury.

BackgroundExercise rehabilitation is demonstrated to improve the prognosis of patients with coronary heart disease (CHD). Statins, as the key medicine to lower cholesterol in CHD, result in skeletal muscle injury and impair exercise training adaptation. Energy metabolism dysfunction is identified as the potential mechanism underlying statin‐induced skeletal muscle injury. In this study, we investigated the effects of the metabolic modulator trimetazidine on skeletal muscle energy metabolism and statin‐associated exercise intolerance.MethodsHigh‐fat fed apolipoprotein E knockout (ApoE−/−) mice were given aerobic exercise and administrated simvastatin, trimetazidine, or simvastatin plus trimetazidine by gavage. Exercise capacity was evaluated at the end of the treatment by hanging grid test, forelimb grip strength, and running tolerance test. Plasma glucose, lipid, and creatine kinase concentrations were measured at the end of the treatment. After sacrifice, gastrocnemii were stored for assessment of muscle morphology and fibre type. Energy metabolism was estimated by plasma lactic acid concentration, ragged red fibres, and glycogen stores. Activities of mitochondrial complex III, citrate synthase activity, and membrane potential were measured to assess mitochondrial function. Oxidative stress was also evaluated by superoxide in mitochondria, superoxide dismutase activity, and glutathione redox state.ResultsIn high‐fat fed ApoE−/− mice, exercise training had no effect on lipid concentrations. Lower lipid concentrations with increased creatine kinase were observed with additional simvastatin treatment. Exercise capacity increased significantly in response to exercise training alone but was blunted by the addition of simvastatin. Similarly, cross‐sectional area of muscle fibres and the proportion of slow‐twitch fibres increased in the exercise group but decreased in the simvastatin plus exercise group. Additionally, simvastatin increased centronucleated fibres and induced energy metabolism dysfunction by inhibiting complex III activity and thus promoted oxidative stress in gastrocnemius. We demonstrated that trimetazidine could reverse simvastatin‐induced exercise intolerance and muscle damages. We also found the ability of trimetazidine in restoration of muscle fibre hypertrophy and facilitating fast‐to‐slow type shift. The energy metabolism dysfunction and oxidative stress in gastrocnemii were rescued by trimetazidine.ConclusionsTrimetazidine alleviated statin‐related skeletal muscle injury by restoration of oxidative phenotype and increasing fibre cross‐sectional areas in response to exercise training. Correspondingly, the exercise training adaptation were improved in high‐fat fed ApoE−/− mice. Moreover, trimetazidine is able to exert its positive effects without affecting the beneficial lipid‐lowering properties of the statins. Thus, trimetazidine could be prescribed to remedy the undesirable statins‐induced exercise intolerance during cardiac rehabilitation in patients with CHD.

Osteogenic/Odontogenic Bioengineering with co-Administration of Simvastatin and Hydroxyapatite on Poly Caprolactone Based Nanofibrous Scaffold.

Purpose: Statin is an effective factor for promoting osteogenesis. The aim of the present study was to evaluate the effect of simvastatin (SIM) and/or HA addition on changes in osteogenesis levels by human DPSCs transferred onto three-dimensional (3D) nanofibrous Poly (ε-caprolactone) (PCL)/Poly lactic acide (PLLA) polymeric scaffolds. Methods: For this purpose, a 3D nanofibrous composite scaffold of PCL/PLLA/HA was prepared by electrospinning method. SIM was added to scaffolds during DPSCs culturing step. Cell proliferation and osteogenic activity levels were assessed by using MTT assay and Alizarin Red assay methods. In addition, the expression of genes responsible for osteogenesis, including BMP2, Osteocalcin, DSPP and RUNX2, were determined before and 2 weeks after incorporation of SIM. Results: The MTT assay showed that PCL/PLLA/HA scaffolds seeded with DPSCs has significant (p<0.05) more proliferative effect than PCL/PLLA or DMEM cultured cells, additionally SIM administration improved this result over the PCL/PLLA/HA scaffolds without SIM treatment. SEM imaging revealed improved adhesion and probably osteogenic differentiation of DPSCs on PCL/PLLA/HA nanofibers treated with SIM, moreover the alizarin red assay ensured significant (p<0.05) higher mineralization of this group. Finally, real time PCR confirmed the positive regulation (P<0.05) of the expression of osteo/odontogenesis markers BMP2, Osteocalcin, DSPP and RUNX2 genes in PLLA-PCL-HA (0.1)-SIM group. Conclusion: As a result, addition of simvastatin with incorporation of hydroxyapatite in PCL-PLLA scaffolds might increase the expression of osteogenesis markers in the DPSCs, with a possible increase in cell differentiation and bone formation.

Simvastatin-Induced Pleuro-Pericardial Effusion and Megaloblastic Anemia

Statins competitively inhibit 3-hyroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase, an enzyme involved in cholesterol synthesis, especially in the liver. Statins are considered for all patients with cardiovascular diseases such as coronary heart disease, peripheral vascular disease, non-haemorrhagic stroke, and transient ischemic attacks. Pleural and pericardial effusions may be caused by infectious diseases, malignancies and autoimmune disorders. However, unexplained effusions should raise the possibility of drugs as being the cause. Drug-induced anemia may be in the form of auto-immune or oxidative hemolysis, aplastic anemia, sideroblastic anemia or megaloblastic anemia. I report a case of lymphocytic pleural effusion, pericardial effusion and megaloblastic anemia in a patient known to have ischemic stroke, diabetes mellitus and hypertension. In the view of un-yielding clinical and laboratory evaluation, a drug-induced reaction was suspected. Because of the recent addition of simvastatin to the patient's regimen, the improvement of the patient's condition after discontinuation of simvastatin and recurrence of symptoms following it's re-administration, a conclusion was made that simvastatin may be the cause of the patient's clinical picture and should be further considered as a cause of unexplained pleural and pericardial effusions, and megaloblastic anemia.

Effect of topical simvastatin (1.2 mg) on gingival crevicular fluid interleukin-6, interleukin-8 and interleukin-10 levels in chronic periodontitis – A clinicobiochemical study

ObjectivesThe impact of simvastatin (SMV), a cholesterol lowering drug, on bone metabolism appears to involve complex interaction with cholesterol metabolites, hormones, inflammatory mediators and growth factors, thus having direct influence on extent and severity of periodontitis. The present study aims to evaluate the in vivo effect of subgingivally delivered SMV gel (1.2mg) as a local drug-delivery agent on clinical parameters and on interleukin-6 (IL-6), interleukin-8 (IL-8) and interleukin-10 (IL-10) levels in gingival crevicular fluid (GCF) of chronic periodontitis patients. Materials and methods50 patients were selected and categorized into two treatment groups: control (scaling and root planing) and test group (scaling and root planing with SMV gel). At initial appointment, clinical parameters were measured. Biochemical analysis of GCF samples was done to evaluate the amount of IL-6, IL-8 and IL-10. GCF sampling and clinical parameters were repeated at one and three months for both the groups. ResultsSMV has an inhibitory effect on pro-inflammatory cytokines (IL-6, IL-8) and stimulatory effect on anti-inflammatory cytokines (IL-10) in GCF of periodontitis patients and has significantly positive effect on all clinical parameters except relative attachment level (RAL). The addition of SMV, thereby, further alters the levels of cytokine that reflect enhanced antibacterial host defence activity at that site. ConclusionTopical SMV has a beneficial effect on periodontal health. Removal of the bacterial plaque and subgingival delivery of SMV significantly modulates the chemokines present in GCF. To summarize, SMV shows promising role in the management of periodontitis.

Successful Treatment of HU-Refractory Polycythemia Vera with Atorvastatin and Low Dose Hydroxyurea. Results from a Pilot Study in Bolivia

BackgroundPolycythemia Vera (PV) is a clonal myeloproliferative neoplasm, characterized by the JAK2V617F mutation.The main goal of current therapies for PV is to prevent thrombotic events and delay transformation to Myelofibrosis (MF) or Acute Myeloid Leukemia (AML).Treatment for PV to keep an hematocrit (Hct) level <45 %, has been associated with a reduction in cardiovascular deaths and thrombotic events (Marchioli, R et al. NEJM 2013). Currently, low-risk PV patients (<60 years and no previous thrombotic events) are treated with aspirin and phlebotomy while high-risk patients require additional cytoreductive therapy, usually with Hydroxyurea (HU). Resistance to HU is associated with an increased risk of transformation and reduced survival. This is why for HU-refractory patients, second line treatments with interferon alpha, anagrelide or even ruxolitinib are recommended. In Latin America, because of high cost and drugs availability, this last group reflects difficulties to be treated. Because statins have been reported to modulate the erythroid clonogenic activity of normal BM erythroid colonies we performed a pilot study to investigate in vitro and in vivo the biologic and clinical activity of atorvastatin in PV patientsPatients and MethodsTen high risk PV patients with a median age of 64.3 years (range 58-73) entered into this study. The diagnosis of PV was done according to the 2008 World Health Organization diagnostic criteria and patients were stratified according to an algorithm proposal provided by Griesshammer et al. (Ann Hematol, 2015). The definition of HU resistance (Barosi, G et al.: BJH 2009) was applicable to five patients (median age 63.9 years) failing to achieve a satisfactory hematologic response upon treatment with more than 2 g of HU, 100 mg of Aspirin and phlebotomies. The assessment of the JAK2V617F mutation was performed as previously described (Guerini et al.: Leukemia 2009). Colony assay, proliferation and apoptosis tests were performed with or without Simvastatin (3.5 uM), as previously described (Amaru, A, Experimental Hematology 2012), on cell lines (UKE1 and K562) and bone marrow mononuclear cells obtained from PV patients and healthy donors. Patients with HU refractory PV (n=5) and high risk PV with hypercholesterolemia (n=5) were eligible to receive Atorvastatin (20 mg/day) added on the top of the ongoing treatment with phlebotomies, Aspirin (100 mg/day) and cytoreductive HU therapy (500 mg/day). All treated patients were high altitude residents (> 3.600 m.a.s.l.) of La Paz (Bolivia) where the normal Hct level of healthy subjects is 48-57% for men and 44-54% for women. This pilot study was approved by the Review Board of the Hospital and the University of San Andres, La Paz.ResultsIn a preliminary set of in vitro proliferation cell assays, simvastatin (3.5 uM), added for 5 days, induced a 33% inhibition of cell proliferation of UKE-1 (JAK2V617F mutated) as compared to 5 % of K562 (BCR/ABL positive). A comparable result was obtained in a 7-day clonogenic cell assay where the colony inhibition was 50 % for UKE-1 and 10 % for K562. On the basis of these results similar experiments were also performed using BM mononuclear cells derived from PV patients and healthy donors. In these experiments performed with the addition of simvastatin, it induced a 41% of inhibition in BFU-E colonies of PV patients and a 25% of inhibition in healthy donors. Furthermore, BFU-E colonies inhibited by simvastatin presented a decrease in hemoglobinization and the size of colonies.HU refractory PV patients and High-risk PV patients with hypercholesterolemia treated with the addition of Atorvastatin, Aspirin, cytoreductive HU and phlebotomies; after a follow-up of 2.6 years (1-7 years), induced a decrease of WBC from 16.500 to 9.270/ul, Hct 61.1 to 52.3% and PLT 457.900,000 to 324.7000/ul. The number of required phlebotomies is reduced in comparison to the required at starting treatment. None of the patients presented thrombotic or cardiopulmonary event. One patient died within two years of starting treatment, due to complications of diabetes mellitus.ConclusionsIn vitro and in vivo, statins showed some evidence of inhibitory activity of the hematopoiesis of PV patients. These preliminary results might indicate the opportunity to further investigate the potential clinical value of these molecules in the treatment of PV. DisclosuresOff Label Use: Atorvastatin was used for its antiproliferative activity on myeloid progenitor cells shown by in vitro experiments.

Perindopril and barnidipine alone or combined with simvastatin on hepatic steatosis and inflammatory parameters in hypertensive patients

The aim of this study was to evaluate the effects of perindopril or barnidipine alone or combined with simvastatin on metabolic parameters and hepatic steatosis degree.One hundred and forty nine mild to moderate hypertensive, normocholesterolemic, overweight or obese outpatients with hepatic steatosis were enrolled. They were treated with perindopril 5mg/day, or barnidipine, 20mg/day, for 6 months; subsequently simvastatin, 20mg/day was added to both treatments for further 6 months. Blood pressure variation was recorded. Patients also underwent an ultrasound examination, at baseline and after 6, and 12 months. We also assessed: fasting plasma glucose (FPG), fasting plasma insulin (FPI), lipid profile, adiponectin (ADN), tumor necrosis factor-α (ΤΝF-α), interleukin-6 (IL-6), high-sensitivity C reactive protein (Hs-CRP).Both perindopril and barnidipine reduced blood pressure, with barnidipine being more effective. Barnidipine, but not perindopril, slightly decreased total cholesterol and triglycerides after 6 months compared to baseline; lipid profile improved in both groups when simvastatin was added. Regarding inflammatory parameters, barnidipine reduced TNF-a, IL-6, and Hs-CRP, both in monotherapy, and after simvastatin addition. Hepatic steatosis parameters improved only when simvastatin was added.We can conclude that barnidipine better reduced blood pressure compared to perindopril and inflammatory parameters. Regarding hepatic steatosis parameters, only the addition of simvastatin improved them.Registration number: NCT02064218, ClinicalTrials.gov

P0105 : Changes in the CNS connexin 43 expression and function in the pathogenesis of hepatic encephalopathy

should also target sinusoidal cells. However, there are no in vitro research tools that correctly mimic the unique features of the sinusoid: adequate spatial distribution of cells, biomechanical stimulation of the endothelial lineage and free paracrine interactions. Our aim was to design, fabricate and validate a three-dimensional co-culture chamber with microfluidics that mimics the hepatic sinusoid. Methods: A transparent PMMA chamber composed by 3 growing areas of 9.7 cm arranged at different heights and separated by 0.5mm was manufactured. The higher level integrated a microfluidic system allowing the application of homogeneous shear stress on a reinforced porous membrane where the endothelium is grown. The middle level had a second culture membrane, and on the lower level, the growing area was the base of the camera. Functional validation: Endothelial cells stimulated with shear stress (5 dyn/cm), HSC, and hepatocytes were cultured for 24h in the chamber. Morphology, viability and endothelial nitric oxide production was analyzed. Translational Validation: The applicability of the chamber in the field of cirrhosis was assessed co-culturing activated HSC and capillarized LSEC under shear stress, or in static conditions. Furthermore, the effects of adding the vasoprotective agent simvastatin were analyzed and compared with traditional culture methods. Results: Functional validation: Cells grown in the chamber showed excellent viability. Endothelial cells were aligned in the direction of shear stress, and increased their production of nitric oxide. Translational Validation: Dysfunctional LSEC cultured in the device showed a marked improvement in their phenotype (activation of the KLF2 pathway and decrease in endothelin-1) that was not observed using ordinary culture methods. The shear stress-derived improvement in LSEC led to a beneficial paracrine effect on HSC (reduced collagen I and alpha-SMA). Simvastatin addition produced a strong protective effect on both cell types, which was significantly higher than that obtained using traditional “transwell” methods. Conclusions: We herein describe a novel, versatile, easy to operate and highly reproducible device that can be applied in different fields of vascular biomedical research, including hepatology.

Simvastatin-Induced Pleuro-Pericardial Effusion and Megaloblastic Anemia

Statins competitively inhibit 3-hyroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase, an enzyme involved in cholesterol synthesis, especially in the liver. Statins are considered for all patients with cardiovascular diseases such as coronary heart disease, peripheral vascular disease, non-haemorrhagic stroke and transient ischemic attacks. Pleural and pericardial effusions may be caused by infectious diseases, malignancies and autoimmune disorders. However, unexplained effusions should raise the possibility of drugs as being the cause. Drug-induced anemia may be in the form of auto-immune or oxidative hemolysis, aplastic anemia, sideroblastic anemia or Megaloblastic anemia. I report a case of lymphocytic pleural effusion, pericardial effusion and Megaloblastic anemia in a patient known to have ischemic stroke, diabetes mellitus and hypertension. In the view of un-yielding clinical and laboratory evaluation, a drug-induced reaction was suspected. Because of the recent addition of Simvastatin to the patient's regimen, the improvement of the patient's condition after discontinuation of Simvastatin and recurrence of symptoms following it's re-administration, a conclusion was made that Simvastatin may be the cause of the patient's clinical picture and should be further considered as a cause of unexplained pleural and pericardial effusions and Megaloblastic anemia.

Statins, 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, potentiate the anti-angiogenic effects of bevacizumab by suppressing angiopoietin2, BiP, and Hsp90α in human colorectal cancer.

Background:Statins, 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, are commonly prescribed because of their therapeutic and preventive effects on cardiovascular diseases. Even though they have been occasionally reported to have antitumour activity, it is unknown whether statins have anti-angiogenic effect in human colorectal cancer (CRC).Methods:A total of 11 human CRC cell lines were used to test the effects of bevacizumab, statins, and bevacizumab plus statins on human umbilical vein endothelial cell (HUVEC) viability and invasion in vitro. To determine the molecular mechanism of statins as anti-angiogenic agents, we performed an angiogenesis antibody array and proteomics analysis and confirmed the results using immunoblot assay, HUVEC invasion rescue assay, and siRNA assay. The antitumoural effects of bevacizumab and statins were evaluated in xenograft models.Results:A conventional dose of statins (simvastatin 0.2 μM, lovastatin 0.4 μM, atorvastatin 0.1 μM, and pravastatin 0.4 μM) in combination with bevacizumab directly reduced the cell viability, migration, invasion, and tube formation of HUVECs. The culture media of the CRC cells treated with bevacizumab or statins were also found to inhibit HUVEC invasion by suppressing angiogenic mediators, such as angiopoietin2, binding immunoglobulin protein (BiP), and Hsp90α. The combined treatment with bevacizumab and simvastatin significantly reduced the growth and metastases of xenograft tumours compared with treatment with bevacizumab alone.Conclusions:The addition of simvastatin at a dose used in patients with cardiovascular diseases (40–80 mg once daily) may potentiate the anti-angiogenic effects of bevacizumab on CRC by suppressing angiopoietin2, BiP, and Hsp90α in cancer cells. A clinical trial of simvastatin in combination with bevacizumab in patients with CRC is needed.

Safety and efficacy of the addition of simvastatin to panitumumab in KRAS mutant metastatic colorectal cancer patients.

e14558 Background: Epithelial growth factor receptor (EGFR) inhibitors are not effective in KRAS mutant colorectal tumors. In these tumors the k-ras protein is activated by post-translational modification by binding farnesyl (C15) and geranylgeranyl (C17) moieties. Both are products of the mevalonate pathway, as is cholesterol. Statins (HMG-CoA-reductase inhibitors) not only inhibit synthesis of cholesterol, but also of C15 and C17 and thus may relevantly inhibit post-translational activation of ras proteins. Therefore, statins may inhibit the expression of the mutant KRAS phenotype and normalize the phenotype into KRAS wild type. In a phase II study we investigated whether simvastatin treatment renders KRAS mutant colorectal tumors sensitive to panitumumab. Methods: Major eligibility criteria were advanced or metastatic colorectal cancer failing prior 5FU, oxaliplatin and irinotecan containing regimens, with a mutation in codon 12, 13 or 61 of KRAS gene on tumor sample. Patients were treated with simvastatin 80mg once daily and panitumumab 6mg/kg every 2 weeks. Primary objective was to investigate the percentage of patients free from progression and alive 11 weeks after the first administration of panitumumab. This phase II study was performed in a Simon two stage design, performing an interim analysis after the enrolment of 15 patients. When at least 6 of these patients (i.e. 40%) were to be alive and free from progression free at 11 weeks after the first administration of panitumumab (similar to the results of the KRAS wild type population of the phase III study), another 31 patients would be included during stage 2. Results: Fifteen evaluable patients were enrolled by 4 study sites. Only one patient was free from progression 11 weeks after start of panitumumab treatment (6.7%). Mean progression free survival was 9.1 weeks (range 5-17 weeks). Most frequent reported side effect was skin toxicity (40%), one patient experienced myositis grade 3. Conclusions: Simvastatin 80mg once daily was not able to inhibit the KRAS mutant phenotype sufficiently to reach sensitivity to panitumumab in colorectal cancer patients with a mutation in the KRAS gene. Clinical trial information: NCT01110785.

Safety and efficacy of the addition of simvastatin to cetuximab in KRAS mutant metastatic colorectal cancer patients.

e14586 Background: Epithelial growth factor receptor (EGFR) inhibitors are not effective in KRAS mutant colorectal tumors. In these tumors the k-ras protein is activated by post-translational modification by binding farnesyl (C15) and geranylgeranyl (C17) moieties. Both are products of the mevalonate pathway, as is cholesterol. Statins (HMG-CoA-reductase inhibitors) not only inhibit synthesis of cholesterol, but also of C15 and C17 and thus may reduce post-translational activation of ras proteins. Therefore, statins may inhibit the expression of the mutant KRAS phenotype and normalize the phenotype into KRAS wild type. In a phase II study we investigated whether simvastatin treatment renders KRAS mutant colorectal tumors sensitive to cetuximab. Methods: Major eligibility criteria were metastatic colorectal cancer failing 5-FU, oxaliplatin and irinotecan containing regimens, with a mutation in codon 12, 13 or 61 of KRAS gene in a tumor sample. Patients were treated with simvastatin 80mg once daily and cetuximab 250mg/m2 weekly (after a starting dose of 400mg/m2). Primary objective was to investigate the percentage of patients free from progression and alive 12.5 weeks after the first administration of cetuximab, similar to the results of the KRAS wild type population of the phase III study. This phase II study was performed in a Simon two stage design, performing an interim analysis after enrolment of 15 patients. If at least 6 of these patients (i.e. 40%) were to be alive and free from progression at 12.5 weeks after the start of cetuximab, another 31 patients would be included during stage II. Results: Fifteen evaluable patients were enrolled at 3 study sites. Four patients (i.e. 27%) were free from progression at time of the primary endpoint. Three patients experienced serious adverse events; an allergic reaction during cetuximab infusion, rhabdomyolysis and intracranial haemorrhage due to cerebral metastases. Conclusions: Though 4 patients experienced stable disease at 12.5 weeks, the predefined efficacy boundary was not met in order to continue enrolment to stage II. Simvastatin 80mg once daily does not induce sensitivity to cetuximab in colorectal cancer patients with a mutation in the KRAS gene. Clinical trial information: NCT01190462.