scholarly journals Intracellular and Plasma Trough Concentration and Pharmacogenetics of Telaprevir

2015 ◽  
Vol 18 (2) ◽  
pp. 171 ◽  
Author(s):  
Jessica Cusato ◽  
Sarah Allegra ◽  
Amedeo De Nicolò ◽  
Lucio Boglione ◽  
Giovanna Fatiguso ◽  
...  
Keyword(s):  
Triple Therapy ◽  
Mononuclear Cells ◽  
Linear Regression Analysis ◽  
Plasma Concentrations ◽  
Kinetic Studies ◽  
Pharmacokinetic Data ◽  
Nucleotide Polymorphisms ◽  
Allelic Discrimination ◽  
Peripheral Blood Mononuclear ◽  
Drug Kinetic

PURPOSE: Triple therapy for HCV-1 infection consists in boceprevir or telaprevir, ribavirin and PEG-interferon. Telaprevir is a P-glycoprotein substrate and it is metabolized by CYP3A4/5. No data have been published on intracellular penetration of telaprevir. We determined peripheral blood mononuclear cells (PBMCs) and trough plasma S and R telaprevir isomers concentrations; moreover, we evaluated the influence of some single nucleotide polymorphisms (SNPs) on these pharmacokinetic data after 1 month of triple therapy in humans. METHODS: Plasma and intracellular telaprevir concentrations were determined at the end of dosing interval (Ctrough) using ULPC-MS/MS validated methods; allelic discrimination was performed through real-time PCR. RESULTS: Median telaprevir Ctrough plasma concentrations were 2579 ng/mL and 2233 ng/mL for the pharmacologically more active S, and R, enantiomers, respectively, with median S/R plasma ratio of 1.11. In PBMC, the medians were 6863 ng/mL and 1096 ng/mL for S and R, respectively, with median S/R being 5.73. The PBMC:plasma ratio for S was 2.59 for R. Plasma ribavirin concentrations were directly correlated with plasma S-telaprevir concentrations. In linear regression analysis, only CYP24A1_rs2585428 SNP (p=0.003) and body mass index (p=0.038) were able to predict S-telaprevir PBMC concentrations. CONCLUSIONS: Our preliminary data could increase the understanding of mechanisms underlying telaprevir intracellular and plasma exposure, suggesting the implementation of pharmacogenetics in these drug kinetic studies. This article is open to POST-PUBLICATION REVIEW. Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.

Abstract 318: Matrix Metalloproteinase 12 is Causally Implicated in Cardiovascular Disease

2016 ◽  
Vol 36 (suppl_1) ◽  
Author(s):  
Ulf Hedin ◽  
Hovsep Mahdessian ◽  
Ljubica Perisic ◽  
Mariette Lengquist ◽  
Karl Gertow ◽  
...  
Keyword(s):  
Cardiovascular Disease ◽  
Matrix Metalloproteinase ◽  
Mononuclear Cells ◽  
Plasma Concentrations ◽  
Intima Media Thickness ◽  
Proximity Ligation Assay ◽  
Nucleotide Polymorphisms ◽  
Peripheral Blood Mononuclear ◽  
Asymptomatic Patients ◽  
The Matrix

Recent evidence suggests that single nucleotide polymorphisms (SNPs) in the matrix metalloproteinase (MMP) gene cluster located at chromosome 11q22.3 are associated with large-vessel stroke. In the present study, we evaluated and extended the reported association by examining the relationship between MMPs and vascular disease in both clinical and experimental samples. Plasma concentrations of MMP-1, MMP-3, MMP-7, MMP-10 and MMP-12 were measured in 3 394 subjects with high-risk for cardiovascular disease (CVD) using the Olink ProSeek CVD array. Plasma MMP-12 concentration showed association with incident cardiovascular events (199 events over 36 months) and intima-media thickness progression over time (p=3.6x10 -5 ). The SNP variant rs1892971 was strongly associated with plasma MMP-12 concentration (p=8x10 -29 ) and weakly with susceptibility to coronary heart disease in the CardiogramplusC4D consortium study (p=8.8x10 -5 ). The same SNP was also significantly associated with MMP-12 gene expression in peripheral blood mononuclear cells using microarrays from patients with carotid atherosclerosis (n=96; p=1.8x10 -4 ). Expression of MMP-12 was strongly increased in carotid plaques (n=127) compared with undiseased arteries (n=10; p<0.0001) and in plaques from symptomatic (n=87) compared to asymptomatic patients (n=40; p=0.03) and localised to CD68+ macrophages. Using proximity ligation assay MMP-12 and elastin was demonstrated to co-interact in plaques in situ, particularly in regions with moderate to strong MMP-12 expression. Silencing of MMP-12 using siRNA in differentiated THP-1 cells indicated that MMP-12 has a role in macrophage migration. In conclusion, our study suggests that MMP-12 is a causal factor in CVD that is highly upregulated in human atherosclerotic plaques where it interacts with elastin and appears to enhance macrophage invasion.

Exercise elevates plasma levels but not gene expression of IL-1β, IL-6, and TNF-α in blood mononuclear cells

2000 ◽  
Vol 89 (4) ◽  
pp. 1499-1504 ◽  
Author(s):  
Andrei I. Moldoveanu ◽  
Roy J. Shephard ◽  
Pang N. Shek
Keyword(s):  
Gene Expression ◽  
Oxygen Uptake ◽  
Mononuclear Cells ◽  
Plasma Concentrations ◽  
Peak Oxygen Uptake ◽  
Cytokine Gene Expression ◽  
Mrna Accumulation ◽  
Peripheral Blood Mononuclear ◽  
Tnf Α ◽  
Blood Mononuclear Cells

Physical activity induces a subclinical inflammatory response, mediated in part by leukocytes, and manifested by elevated concentrations of circulating proinflammatory cytokines, including interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α). However, the source of the cytokines that appear during exercise remains unknown. In this study, we examined exercise-induced changes in plasma cytokine concentrations and their corresponding mRNA expression in peripheral blood mononuclear cells. Ten healthy [peak oxygen uptake = 48.8 ± 6.5 (SD) ml · kg−1 · min−1] but untrained men [age = 25 ± 5 (SD) yr] undertook 3 h of exercise (cycling and inclined walking) at 60–65% peak oxygen uptake. Circulating leukocyte subset counts were elevated during and 2 h postexercise but returned to normal within 24 h. Plasma concentrations of IL-1β, IL-6, and TNF-α peaked at the end of exercise and remained elevated at 2 h (IL-6) and up to 24 h (IL-1β and TNF-α) postexercise. Cytokine gene expression in circulating mononuclear cells was measured by using the reverse transcriptase-polymerase chain reaction; mRNA accumulation did not change with exercise. In conclusion, mRNA accumulation of IL-1β, IL-6, and TNF-α in circulating mononuclear cells is not affected by 3 h of moderate endurance exercise and does not seem to account for the observed increases in plasma cytokines.

Dose-escalation and pharmacodynamic study of topotecan in combination with cyclophosphamide in patients with refractory cancer.

1997 ◽  
Vol 15 (1) ◽  
pp. 148-157 ◽  
Author(s):  
J R Murren ◽  
S Anderson ◽  
J Fedele ◽  
G Pizzorno ◽  
D Belliveau ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Mononuclear Cells ◽  
Day Treatment ◽  
Alkylating Agents ◽  
Plasma Concentrations ◽  
Fixed Dose ◽  
Sign Test ◽  
Published Data ◽  
Drug Induced ◽  
Peripheral Blood Mononuclear

PURPOSE Based on preclinical data that demonstrated synergy between alkylating agents and topoisomerase (topo) I poisons, we determined the maximum-tolerated dose (MTD) of topotecan, using a 5 day bolus schedule, that could be given in combination with a single, fixed dose of cyclophosphamide. Pharmacodynamics of this combination were explored by analyzing biochemical effects of treatment in peripheral-blood mononuclear cells (PBMCs). PATIENTS AND METHODS Patients with refractory cancer were treated with cyclophosphamide 600 mg/m2 on day 1, followed by topotecan given as a 30-minute infusion for 5 consecutive days. Cycles were repeated every 3 weeks. Once the MTD was defined, granulocyte colony-stimulating factor (G-CSF) was added to the regimen in an attempt to escalate further the dose of topotecan. Plasma concentrations of topotecan were determined during the first treatment cycle by high-performance liquid chromatography. PBMCs were sampled at baseline and throughout the 5-day treatment period for analysis of topo I protein concentrations and to determine drug-induced DNA fragmentation. RESULTS Twenty-six patients were treated with topotecan at doses that ranged from 0.5 mg/m2/d to 1.2 mg/ m2/d for a total of 74 cycles. Reversible neutropenia was dose-limiting, with mild to moderate suppression of the other blood-cell elements commonly occurring. Transfusions of RBCs and platelets were required in 24% and 7% of treatment cycles, respectively. The most prominent nonhematologic toxicities were fatigue and weight loss. Compared with previously published data in which topotecan was administered alone, cyclophosphamide did not appear to alter the pharmacokinetics of topotecan. Significant increases in topo I concentration were identified in PBMCs following the administration of cyclophosphamide on day 1 and there was a significant decrease in topo 1 during the 5-day course of treatment (P < .01, sign test). DNA fragmentation as a result of drug treatment was identified in 11 of 15 (73%) cycles analyzed. CONCLUSION For previously treated patients, the recommended dose of topotecan in this schedule is 0.75 mg/m2/d without growth factor support and 1.0 mg/ m2/d if it is administered with G-CSF. Biochemical changes in cells induced by exposure to camptothecins can be measured in vivo and these effects may have important implication in the design of combination therapies and the optimal scheduling of this class of agents.

NK-Cell Reconstitution after HLA-Identical Blood and Marrow Transplantations for CML: The Recovery of NKG2D Is Inversely Correlated with NKG2A and It Promotes the GVL Effect.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4879-4879
Author(s):  
Juan Tong ◽  
Huilan Liu ◽  
Liangquan Geng ◽  
Zimin Sun ◽  
Baolin Tang ◽  
...  
Keyword(s):  
Nk Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Nk Cell ◽  
Linear Regression Analysis ◽  
Cell Activity ◽  
Target Cells ◽  
Peripheral Blood Stem Cells ◽  
Peripheral Blood Mononuclear ◽  
Blood Stem Cells

Abstract Natural killer (NK) cell alloreactivity is reported to mediate strong graft versus leukemia (GVL) effect in patients after allogeneic stem-cell transplantation. NKG2D receptors recognize human MHC class Ichain related A and B (MICA/B) and UL16-binding protein 1∼4(ULBP 1∼4) on target cells, thereby regulating NK cell activity. To examine the recovery of NKG2D, NKG2A and other receptors expression by NK cells, we used flow cytometry to evaluate samples from 11 chronic myeloid leukemia patients and their donors in the year following unmanipulated HLA completely matched peripheral blood stem cells plus bone marrow transplantation. Peripheral blood mononuclear cells from patients and their donors were tested in standard 51Cr release assays against cultured K562 targets to determine the cytotoxicity of the NK cells in the same intervals. There is no mismatched immunoglobulin-like receptor (KIR) ligand in both GVH and HVG direction. The reconstitution of KIR2DL1 (CD158a) after this transplantation protocol was very slow and these receptors didn’t reach normal value in the year and KIR2DL2 (CD158b) was much better. The NKG2D increased and the NKG2A decreased quickly at the same time after engraftment, and used linear regression analysis we demonstrated that NKG2A recovery was inversely correlated with NKG2D recovery in the year following transplantation. The ratio of NKG2D/NKG2A was directly associated with the capacity of NK-cell cytotoxicity. Thus, the reconstitution of NKG2D makes contribution to the recovery of the NK cytotoxicity. These results reveals that the NK cells generated after HLA matched blood plus bone morrow transplantation of CML patients are promoted at an immature state characterized by specific phenotypic features and enhanced functioning, having potential impact for immune responsiveness and transplantation outcome.

Genetic variations in the SOCS3 gene in patients with Graves’ ophthalmopathy

2015 ◽  
Vol 68 (6) ◽  
pp. 448-452 ◽  
Author(s):  
Ruijia Yan ◽  
Junjie Yang ◽  
Ping Jiang ◽  
Ling Jin ◽  
Jing Ma ◽  
...  
Keyword(s):  
Western Blot ◽  
Mononuclear Cells ◽  
Pcr Analysis ◽  
Nucleotide Polymorphisms ◽  
Chinese Han ◽  
Peripheral Blood Mononuclear ◽  
Protein Levels ◽  
Qrt Pcr ◽  
Graves Ophthalmopathy ◽  
Socs3 Mrna

AimsTo explore the role of the suppressor of cytokine signalling 3 (SOCS3) gene in Graves’ ophthalmopathy (GO) patients.MethodsA case–control study was conducted in a Chinese Han population by recruiting 114 Graves’ disease (GD) patients with GO and 156 GD patients without GO. We determined SOCS3 mRNA and protein levels in Epstein–Barr virus-transformed lymphoblastoid cell lines (EBV-LCLs) from peripheral blood mononuclear cells (PBMCs) by quantitative real-time (QRT)-PCR analysis and western blot analysis. We also genotyped five single nucleotide polymorphisms (SNPs) in the SOCS3 locus (SOCS3 rs12952093, rs4969170, rs4969168, rs4969169 and rs2280148) in all 270 GD patients using ligase detection reaction and multiplex PCR analyses. QRT-PCR and western blot assays were then performed to compare SOCS3 mRNA and protein levels between the rs4969170 AA and GG genotype groups from 20 GO patients.ResultsBasal SOCS3 mRNA and protein expression levels were significantly increased in patients with GO (p<0.05). The SOCS3 rs4969170 AA genotype was strongly associated with GO (OR=3.5, 95% CI 1.6 to 7.5, p=0.001). The AA genotype carriers had significantly higher SOCS3 mRNA and protein levels than those with the GG genotype (p<0.05).ConclusionsPatients with GD who carry the AA genotype of the rs4969170 SNP in SOCS3 are more susceptible to the development of GO.

scholarly journals Differential Expression of Apoptotic and Low-Grade Inflammatory Markers in Alzheimer Disease Compared to Diabetes Mellitus Type 1 and 2

2019 ◽  
Vol 3 (6) ◽  
pp. 1003-1013 ◽  
Author(s):  
Krystallenia I Alexandraki ◽  
Nikolaos V Apostolopoulos ◽  
Christos Adamopoulos ◽  
Evangelia Stamouli ◽  
Georgia Dalagiorgou ◽  
...  
Keyword(s):  
Alzheimer Disease ◽  
Fas Ligand ◽  
Mononuclear Cells ◽  
Plasma Concentrations ◽  
Diabetes Mellitus Type 1 ◽  
Low Grade ◽  
Peripheral Blood Mononuclear ◽  
Reactive Protein ◽  
Before And After

Abstract Background Neuroinflammation, impaired brain insulin signaling, and neuronal apoptosis may be interrelated in the pathophysiology of people with Alzheimer disease (AD) and diabetes, either type 1 or 2 diabetes (T1D or T2D, respectively). Methods We studied 116 patients: 41 with AD, 20 with T1D, 21 with T2D, and 34 healthy controls. The number (n) of cytokine-secreting peripheral blood mononuclear cells (PBMCs) before and after mitogenic stimulation was determined for interleukin 1β (IL1β), interleukin 6 (IL6), tumor necrosis factor (TNF) by the enzyme-linked-immuno-spot assay. Serum concentrations of C-reactive protein (CRP) and Fas ligand (FASLG) were determined by ELISA. Results The studied subgroups did not differ in sex but differed in age. Higher CRP concentrations were detected in the AD group than in the T1D group (P = 0.02) and lower in controls (P &lt; 0.001). The nPBMCs was higher in AD patients after stimulation than in basal conditions: after stimulation in nTNF (P &lt; 0.001 vs T2D; P &lt; 0.001 vs T1D; P = 0.001 vs control), nIL6 (P = 0.039 vs T2D; P &lt; 0.001 vs T1D; P = 0.007 vs control), and nIL1β (P = 0.03 vs control). The nPBMCs increased after stimulation with ΡΜA in all the subgroups (P &lt; 0.001). FASLG in the AD group displayed statistically higher concentrations than in all other subgroups (P &lt; 0.001 vs T2D; P &lt; 0.001 vs T1D; P = 0.012 vs control). The nPBMCs was positively correlated with plasma concentrations of FASLG in the AD subgroup. Conclusions Patients with AD display a low-grade systemic inflammation compared to people with diabetes. The FAS–FASLG pathway has a potential role because FASLG concentrations are positively correlated with the inflammatory response in AD. However, this positive correlation cannot be seen in people with diabetes, at least not with the apoptotic markers used in the present study.

scholarly journals MAPK pathway and B cells overactivation in multiple sclerosis revealed by phosphoproteomics and genomic analysis

2019 ◽  
Vol 116 (19) ◽  
pp. 9671-9676 ◽  
Author(s):  
Ekaterina Kotelnikova ◽  
Narsis A. Kiani ◽  
Dimitris Messinis ◽  
Inna Pertsovskaya ◽  
Vicky Pliaka ◽  
...  
Keyword(s):  
Multiple Sclerosis ◽  
B Cells ◽  
Kinase Inhibitors ◽  
Mononuclear Cells ◽  
Mapk Pathway ◽  
Genomic Analysis ◽  
Nucleotide Polymorphisms ◽  
Peripheral Blood Mononuclear ◽  
Disease Modifying Drugs

Dysregulation of signaling pathways in multiple sclerosis (MS) can be analyzed by phosphoproteomics in peripheral blood mononuclear cells (PBMCs). We performed in vitro kinetic assays on PBMCs in 195 MS patients and 60 matched controls and quantified the phosphorylation of 17 kinases using xMAP assays. Phosphoprotein levels were tested for association with genetic susceptibility by typing 112 single-nucleotide polymorphisms (SNPs) associated with MS susceptibility. We found increased phosphorylation of MP2K1 in MS patients relative to the controls. Moreover, we identified one SNP located in the PHDGH gene and another on IRF8 gene that were associated with MP2K1 phosphorylation levels, providing a first clue on how this MS risk gene may act. The analyses in patients treated with disease-modifying drugs identified the phosphorylation of each receptor’s downstream kinases. Finally, using flow cytometry, we detected in MS patients increased STAT1, STAT3, TF65, and HSPB1 phosphorylation in CD19+ cells. These findings indicate the activation of cell survival and proliferation (MAPK), and proinflammatory (STAT) pathways in the immune cells of MS patients, primarily in B cells. The changes in the activation of these kinases suggest that these pathways may represent therapeutic targets for modulation by kinase inhibitors.

A first-in-man phase I study of R306465, a histone deacetylase (HDAC) inhibitor exploring pharmacokinetics (PK) and pharmacodynamics (PD) utilizing an electrochemiluminescent immunoassay in patients (p) with advanced tumours

2007 ◽  
Vol 25 (18_suppl) ◽  
pp. 3578-3578 ◽  
Author(s):  
P. C. Fong ◽  
S. Settatree ◽  
R. Sinha ◽  
A. Hardcastle ◽  
P. W. Hellemans ◽  
...  
Keyword(s):  
Mononuclear Cells ◽  
Plasma Concentrations ◽  
Hdac Inhibitors ◽  
Fold Increase ◽  
Maximum Tolerated Dose ◽  
Transcriptional Level ◽  
Peripheral Blood Mononuclear ◽  
Fed State ◽  
Class 1 ◽  
Ecog Ps

3578 Background: R306565 is an aromatic hydroxamic acid with predominant inhibitory effects on Class 1 HDAC enzymes (with IC50 ∼10 nM). HDAC inhibitors (HDACi) affect gene expression at the transcriptional level, leading to cell cycle arrest and induction of apoptosis. Methods: P with solid tumours or lymphoma were given R306465 orally daily for 3 weeks (w) out of 4 in an escalating schedule. Objectives include safety, tolerability, PK (including food effect exploration), PD evaluation and circulating tumour cell (CTC) quantitation. Results: Four dose levels (100, 200, 300 and 400 mg) have been evaluated involving 15 p (7 male), age range 29–72 (median 59 y) and ECOG PS 0–2. A total of 37 cycles have been administered. Most common adverse events (AE) were Grade (G) 1–3 fatigue (87%), G1–2 nausea (66%), G1–2 vomiting (33%), G1–2 diarrhoea (40%), and G1–2 anorexia (40%). Dose limiting toxicity of G3 fatigue was seen in 1/6 p in the 400mg cohort. PK parameters were approximately dose proportional. Plasma concentrations increased in the fed state. PD effect of histone H3 acetylation (AcH3) in peripheral blood mononuclear cells (PBMC) was determined quantitatively with a novel validated electrochemiluminescent immunoassay developed in-house (applying Mesoscale Discovery technology). Although some interpatient variability exists, increased AcH3 was observed in 2/6 p in the 400 mg cohort, while the percentage rise in AcH3 was minimal for cohorts 1–3. Peak AcH3 achieved in 2 p dosed at 400 mg was approximately 5–10 fold increase over baseline. Using CellSearch technology for quantitation of CTCs, 8/14 p had detectable CTCs at baseline; the CTC trend will be presented. 4 p had stable disease (SD) for = 4 months. Conclusions: R306465 could be safely administered on a daily dosing schedule for 3 of 4 w up to 400 mg. Common toxicities seen were gastrointestinal and fatigue. Maximum tolerated dose has not been reached. PK suggests dose proportionality. Promising PD data showing increased acetylation in PBMC at 400 mg, further supports the utilization of the immunoassay platform in HDACi clinical trials. No significant financial relationships to disclose.

scholarly journals Population Pharmacokinetic Modeling of Plasma and Intracellular Ribavirin Concentrations in Patients with Chronic Hepatitis C Virus Infection

2015 ◽  
Vol 59 (4) ◽  
pp. 2179-2188 ◽  
Author(s):  
Liviawati S. Wu ◽  
Joseph E. Rower ◽  
James R. Burton ◽  
Peter L. Anderson ◽  
Kyle P. Hammond ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Mononuclear Cells ◽  
Formation Rate ◽  
Plasma Concentrations ◽  
Compartment Model ◽  
Population Pharmacokinetic ◽  
Peripheral Blood Mononuclear ◽  
First Order

ABSTRACTRibavirin, a guanosine analog, is a broad-spectrum antiviral agent. Ribavirin has been a fundamental component of the treatment of hepatitis C virus (HCV) infection for decades, but there is a very limited understanding of the clinical pharmacology of this drug. Furthermore, it is associated with a major dose-limiting toxicity, hemolytic anemia. Ribavirin undergoes intracellular phosphorylation by host enzymes to ribavirin monophosphate (RMP), ribavirin diphosphate (RDP), and ribavirin triphosphate (RTP). The intracellular forms have been associated with antiviral and toxic effectsin vitro, but the kinetics of these phosphorylated moieties have not been fully elucidatedin vivo. We developed a model to characterize the plasma pharmacokinetics of ribavirin and the difference between intracellular phosphorylation kinetics in red cells (nonnucleated) and in peripheral blood mononuclear cells (nucleated). A time-independent two-compartment model with first-order absorption described the plasma data well. The cellular phosphorylation kinetics was described by a one-compartment model for RMP, with the formation rate driven by plasma concentrations and the first-order degradation rate. RDP and RTP rapidly reached equilibrium with RMP. Concomitant telaprevir use, inosine triphosphatase genetics, creatinine clearance, weight, and sex were significant covariates. The terminal ribavirin half-life in plasma and phosphorylated anabolites in cells was approximately 224 h. We found no evidence of time-dependent kinetics. These data provide a foundation for uncovering concentration-effect associations for ribavirin and determining the optimal dose and duration of this drug for use in combination with newer direct-acting HCV agents. (This study has been registered at ClinicalTrials.gov under registration no. NCT01097395.)

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