scholarly journals Differences in Mammalian Target of Rapamycin Gene Expression in the Peripheral Blood and Articular Cartilages of Osteoarthritic Patients and Disease Activity

Arthritis ◽  
2013 ◽  
Vol 2013 ◽  
pp. 1-14 ◽  
Author(s):  
Elena V. Tchetina ◽  
A. Robin Poole ◽  
Elena M. Zaitseva ◽  
Eugeniya P. Sharapova ◽  
Natalya G. Kashevarova ◽  
...  
Keyword(s):  
Gene Expression ◽  
Disease Activity ◽  
Peripheral Blood ◽  
Caspase 3 ◽  
Joint Destruction ◽  
Mammalian Target Of Rapamycin ◽  
Joint Stiffness ◽  
P70 S6k ◽  
Articular Cartilages ◽  
End Stage

The gene expression of mTOR, autophagy-related ULK1, caspase 3, CDK-inhibitor p21, and TNFα was measured in the peripheral blood of osteoarthritic (OA) patients at different stages of the disease aiming to establish a gene expression profile that might indicate the activity of the disease and joint destruction. Whole blood of 65 OA outpatients, 27 end-stage OA patients, 27 healthy volunteers, and knee articular cartilages of 28 end-stage OA patients and 26 healthy subjects were examined. OA outpatients were subjected to clinical testing, ultrasonography, and radiographic and WOMAC scoring. Protein levels of p70-S6K, p21, and caspase 3 were quantified by ELISA. Gene expression was measured using real-time RT-PCR. Upregulation of mTOR gene expression was observed in PBMCs of 42 OA outpatients (“High mTOR expression subset”) and in PBMCs and articular cartilages of all end-stage OA patients. A positive correlation between mTOR gene expression in PBMCs and cartilage was observed in the end-stage OA patients. 23 OA outpatients in the “Low mTOR expression subset” exhibited significantly lower mTOR gene expression in PBMCs compared to healthy controls. These “Low mTOR” subset subjects experienced significantly more pain upon walking, and standing and increased total joint stiffness versus “High mTOR” subset, while the latter more often exhibited synovitis. The protein concentrations of p70-S6K, p21, and caspase 3 in PBMCs were significantly lower in the “Low” subset versus “High” subset and end-stage subjects. Increases in the expression of mTOR in PBMCs of OA patients are related to disease activity, being associated with synovitis more than with pain.

scholarly journals Association of Bone Loss with the Upregulation of Survival-Related Genes and Concomitant Downregulation of Mammalian Target of Rapamycin and Osteoblast Differentiation-Related Genes in the Peripheral Blood of Late Postmenopausal Osteoporotic Women

2015 ◽  
Vol 2015 ◽  
pp. 1-8 ◽  
Author(s):  
Elena V. Tchetina ◽  
Karina A. Maslova ◽  
Mikhail Y. Krylov ◽  
Valery A. Myakotkin
Keyword(s):  
Gene Expression ◽  
Bone Loss ◽  
Peripheral Blood ◽  
Osteoblast Differentiation ◽  
Molecular Mechanisms ◽  
Caspase 3 ◽  
Mammalian Target Of Rapamycin ◽  
Target Of Rapamycin ◽  
Healthy Controls ◽  
Mineral Density

We aimed to identify bone related markers in the peripheral blood of osteoporotic (OP) patients that pointed toward molecular mechanisms underlying late postmenopausal bone loss. Whole blood from 22 late postmenopausal OP patients and 26 healthy subjects was examined. Bone mineral density (BMD) was measured by DXA. Protein levels of p70-S6K, p21, MMP-9, TGFβ1, and caspase-3 were quantified by ELISA. Gene expression was measured using real-time RT-PCR. OP registered by low BMD indices in late postmenopausal patients was associated with a significant upregulation of autophagy proteinULK1, cyclin-dependent kinase inhibitorp21, and metalloproteinaseMMP-9gene expression in the blood compared to the healthy controls and in a significant downregulation ofmTOR(mammalian target of rapamycin),RUNX2, andALPLgene expression, while expression ofcathepsin K, caspase-3, transforming growth factor(TGF)β1, interleukin-(IL-)1β, andtumor necrosis factorα(TNFα) was not significantly affected. We also observed a positive correlation betweenTGFβ1andRUNX2expression and BMD at femoral sites in these patients. Therefore, bone loss in late postmenopausal OP patients is associated with a significant upregulation of survival-related genes (ULK1andp21) andMMP-9, as well as the downregulation ofmTORand osteoblast differentiation-related genes (RUNX2andALPL) in the peripheral blood compared to the healthy controls.

scholarly journals THU0472 CATHEPSIN S GENE EXPRESSION MEASURED IN THE PERIPHERAL BLOOD OF OSTEOARTHRITIC PATIENTS PRIOR TO SURGERY AS A BIOMARKER OF POST-OPERATIVE PAIN DEVELOPMENT

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 473.2-473
Author(s):  
E. Tchetina ◽  
K. Glemba ◽  
G. Markova ◽  
E. Taskina ◽  
M. Makarov
Keyword(s):  
Gene Expression ◽  
Postoperative Pain ◽  
Joint Replacement ◽  
Peripheral Blood ◽  
Caspase 3 ◽  
Cathepsin S ◽  
Joint Replacement Surgery ◽  
Replacement Surgery ◽  
Post Operative Pain ◽  
End Stage

Background:Osteoarthritis (OA) is a chronic rheumatic disease, which involves pain, limited inflammation, and local destruction of the knee joint. OA pain is a major clinical symptom, which limits working capacity and denotes an important indication for joint replacement in the end-stage OA. In spite of significant number of positive outcomes, chronic postoperative pain represents a major adverse consequence of surgery, which is observed in 10-40% of OA patients. Therefore, identification of patients potentially capable of developing chronic postoperative pain prior to surgery could significantly improve therapy outcome. Recently we hypothesized that genes related to pain sensitization whose expression is upregulated in about 10-40% of the examined end-stage OA patient cohort might be responsible for postoperative pain. Retrospective analysis of gene expression in the peripheral blood of end-stage OA patients before joint replacement surgery revealed that expression of cathepsins S and K, caspase 3, and MMP-9 genes might be associated with postoperative pain development [Ann Rheum Dis,78, suppl 2:A520].Objectives:To examine the validity of our hypothesis in the prospective study.Methods:We examined peripheral blood of 26 healthy volunteers (average age 55±8.3 years old) and 40 end-stage OA patients (average age 56.5±8.9 years old) undergoing joint replacement surgery. Patients were examined before and 6 months after surgery. Pain was assessed prior to surgery using VAS index and neuropathic pain questionnaires DN4 and PainDETECT. Functional activity was evaluated by WOMAC. After surgery pain indices according to VAS of 30% and higher were considered. MMP-9 and caspase 3 protein levels were quantified by ELISA. Total RNA isolated from whole blood was used in expression studies for caspase 3; metalloproteinase (MMP)-9; cathepsins K and S genes. These were performed with quantitative real-time RT-PCR.Results:Out of 40 patients pain complaints were obtained from 9 patients (22,5%) after 6 months’ post-surgery. Prior to surgery all the examined genes were significantly upregulated in the patients who developed post-operative pain compared to healthy controls and those subjects who did not develop pain after surgery. However, no difference in the levels of the examined pain-related and functional indices in patients, who developed pain or not, was noted before surgery. ROC curve analyses confirmed significant associations (p<0.05) between expressions of the examined genes prior to surgery with the likelihood of pain development after surgery. The cut-off values for the examined gene expressions were 11.34 for cathepsin S (sensitivity of 0.89 and specificity of 0.76), 10.11 for caspase 3 (sensitivity of 0.86 and specificity of 0.65), 10.09 for cathepsin K (sensitivity of 0.86 and specificity of 0.78). Moreover, among the examined genes cathepsin S expression was the most informative predictor of postoperative pain development [AUC= 0.857, 95%CI (0.708-1.000)].Conclusion:High cathepsin S gene expression in the peripheral blood of the end-stage OA patients measured prior to joint replacement surgery could serve an important biomarker of post-operative pain development.Disclosure of Interests: :None declared

scholarly journals Rheumatoid Factor Positivity Is Associated with Increased Joint Destruction and Upregulation ofMatrix Metalloproteinase 9andCathepsin KGene Expression in the Peripheral Blood in Rheumatoid Arthritic Patients Treated with Methotrexate

2013 ◽  
Vol 2013 ◽  
pp. 1-10 ◽  
Author(s):  
Elena V. Tchetina ◽  
Natalia V. Demidova ◽  
Dmitry E. Karateev ◽  
Eugeny L. Nasonov
Keyword(s):  
Gene Expression ◽  
Rheumatoid Factor ◽  
Peripheral Blood ◽  
Caspase 3 ◽  
Joint Destruction ◽  
Previous History ◽  
Cathepsin K ◽  
Tissue Destruction ◽  
Rheumatoid Arthritic

We evaluated changes in gene expression ofmTOR,p21,caspase-3,ULK1,TNFα, matrix metalloproteinase(MMP)-9, andcathepsin Kin the whole blood of rheumatoid arthritic (RA) patients treated with methotrexate (MTX) in relation to their rheumatoid factor status, clinical, immunological, and radiological parameters, and therapeutic response after a 24-month follow-up. The study group consisted of 35 control subjects and 33 RA patients without previous history of MTX treatment. Gene expression was measured using real-time RT-PCR. Decreased disease activity in patients at the end of the study was associated with significant downregulation ofTNFαexpression. Downregulation ofmTORwas observed in seronegative patients, while no significant changes in the expression ofp21,ULK1, orcaspase-3were noted in any RA patients at the end of the study. The increase in erosion numbers observed in the seropositive patients at the end of the follow-up was accompanied by upregulation ofMMP-9andcathepsin K, while seronegative patients demonstrated an absence of significant changes inMMP-9andcathepsin Kexpression and no increase in the erosion score. Our results suggest that increased expression ofMMP-9andcathepsin Kgenes in the peripheral blood might indicate higher bone tissue destruction activity in RA patients treated with methotrexate. The clinical study registration number is 0120.0810610.

scholarly journals Development of Postoperative Pain in Patients with End-Stage Knee Osteoarthritis Is Associated with Upregulation of Genes Related to Extracellular Matrix Degradation, Inflammation, and Apoptosis Measured in the Peripheral Blood before Knee Surgery

Life ◽  
2020 ◽  
Vol 10 (10) ◽  
pp. 224
Author(s):  
Elena V. Tchetina ◽  
Kseniya E. Glemba ◽  
Galina A. Markova ◽  
Evgeniy A. Naryshkin ◽  
Elena A. Taskina ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Postoperative Pain ◽  
Joint Replacement ◽  
Peripheral Blood ◽  
Caspase 3 ◽  
Mononuclear Cells ◽  
Tissue Inhibitor Of Metalloproteinase ◽  
Gene Expressions ◽  
Joint Replacement Surgery ◽  
End Stage

Osteoarthritis (OA) pain implies an indication for joint replacement in patients with end-stage OA. However, chronic postoperative pain is observed in 10–40% of patients with OA. Here, we identified genes whose expression in the peripheral blood before surgery could denote the risk of postoperative pain development. We examined the peripheral blood of 26 healthy subjects and 50 patients with end-stage OA prior to joint replacement surgery. Pain was evaluated before surgery using the visual analog scale (VAS) index and neuropathic pain questionnaires, Douleur Neuropathique 4 Questions (DN4) and PainDETECT questionnaires. Functional activity was assessed using the Western Ontario and McMaster Universities osteoarthritis index (WOMAC). Three and six months after surgery, pain indices according to VAS of 30% and higher were considered. Metalloproteinase (MMP)-9 and tissue inhibitor of metalloproteinase (TIMP)1 protein levels were measured using ELISA in the peripheral blood mononuclear cells (PBMCs). Total RNA isolated from whole blood was analysed using quantitative real-time RT-PCR for caspase-3, MMP-9, TIMP1, cathepsins K and S, tumour necrosis factor (TNF)α, interleukin (IL)-1β, and cyclooxygenase (COX)-2 gene expression. Seventeen patients reported post-surgical pain. Expression of cathepsins K and S, caspase-3, TIMP1, IL-1β, and TNFα genes before surgery was significantly higher in these patients compared to pain-free patients with OA. Receiver-operating characteristic (ROC) curve analyses confirmed significant associations between these gene expressions and the likelihood of pain development after arthroplasty. High baseline expression of genes associated with extracellular matrix destruction (cathepsins S and K, TIMP1), inflammation (IL-1β, TNFα), and apoptosis (caspase-3) measured in the peripheral blood of patients with end-stage OA before knee arthroplasty might serve as an important biomarker of postoperative pain development.

Peripheral Blood Expression Profiles of Bone Morphogenetic Proteins, Tumor Necrosis Factor-superfamily Molecules, and Transcription Factor Runx2 Could Be Used as Markers of the Form of Arthritis, Disease Activity, and Therapeutic Responsiveness

2009 ◽  
Vol 37 (2) ◽  
pp. 246-256 ◽  
Author(s):  
DANKA GRCEVIC ◽  
ZRINKA JAJIC ◽  
NATAŠA KOVACIC ◽  
IVAN KREŠIMIR LUKIC ◽  
VEDRAN VELAGIC ◽  
...  
Keyword(s):  
Gene Expression ◽  
Tumor Necrosis Factor ◽  
Disease Activity ◽  
Peripheral Blood ◽  
Tumor Necrosis ◽  
Fas Ligand ◽  
Expression Profiles ◽  
Quantitative Polymerase Chain Reaction ◽  
Roc Curve Analysis ◽  
Necrosis Factor

Objective.To assess whether different forms of arthritis and disease activity could be distinguished by peripheral blood expression profiles of bone-regulatory factors including tumor necrosis factor (TNF)-superfamily [TNF-related apoptosis-inducing ligand (TRAIL), the Fas ligand (FasL), and the ligand for herpesvirus entry mediator (LIGHT)] and bone morphogenetic protein (BMP)-family members (BMP-2, BMP-4, BMP-6) as well as osteoblast differentiation gene Runx2.Methods.Blood cells from healthy controls (n = 25) and patients at different disease stages with rheumatoid arthritis (RA; n = 49), osteoarthritis (OA; n = 17), or spondyloarthritis, including ankylosing spondylitis (AS; n = 27) or psoriatic arthritis (PsA; n = 23), were processed for quantitative polymerase chain reaction. Gene expression was assessed in comparison with control samples, correlated with clinical data of different forms of arthritis, and analyzed for discriminative efficacy between groups by receiver-operation characteristic (ROC) curves. Results were confirmed on diagnostic RA (n = 5) and AS (n = 8) samples.Results.BMP-4, BMP-6, and Runx2 expressions were significantly decreased in patients with RA and OA versus controls. Patients with RA also had decreased FasL and LIGHT expression, while patients with AS had increased Runx2 expression. Negative correlation with disease activity was found for BMP-4, FasL, and Runx2 in RA and for Runx2 in PsA, while positive correlation was found for BMP-4 in PsA. Gene expression was higher in the therapy-resistant form of AS (for BMP-4, LIGHT, and Runx2) and in methotrexate-treated patients in RA (for BMP-2 and LIGHT). ROC curve analysis confirmed discrimination between groups, particularly decreased LIGHT and Runx2 for RA and increased Runx2 for AS.Conclusion.Our study identified BMP and Runx2 as possible biomarkers of bone metabolism in several forms of arthritis, while lower FasL and LIGHT were associated with RA. Correlation between gene expression and disease activity may be clinically useful in assessing therapeutic effectiveness and disease monitoring.

Gene expression profiling in peripheral blood nuclear cells in patients with refractory ischaemic end-stage heart failure

2010 ◽  
Vol 51 (3) ◽  
pp. 353-368 ◽  
Author(s):  
S. Szmit ◽  
M. Jank ◽  
H. Maciejewski ◽  
M. Grabowski ◽  
R. Glowczynska ◽  
...  
Keyword(s):  
Gene Expression ◽  
Heart Failure ◽  
Gene Expression Profiling ◽  
Peripheral Blood ◽  
Expression Profiling ◽  
End Stage Heart Failure ◽  
End Stage

scholarly journals Metabolic aspects of clinical remission prediction from baseline blood gene expression in patients with rheumatoid arthritis treated with methotrexate

2019 ◽  
Vol 13 (2) ◽  
pp. 47-54
Author(s):  
E. V. Chetina ◽  
N. V. Demidova ◽  
G. A. Markova
Keyword(s):  
Rheumatoid Arthritis ◽  
Gene Expression ◽  
Disease Activity ◽  
Clinical Remission ◽  
Structural Changes ◽  
Caspase 3 ◽  
Unknown Etiology ◽  
Control Group ◽  
Healthy Individuals ◽  
Number Of Patients

Rheumatoid arthritis (RA) is an autoimmune disease of unknown etiology, which is characterized by chronic erosive arthritis (synovitis) and systemic inflammation of the viscera. Methotrexate (MTX) is the drug of choice for RA treatment. However, it is currently impossible to predict the efficacy of MTX in a particular patient; the drug fails to produce the desired effect or causes adverse reactions in a considerable number of patients. The identification of patients who are responsive to MTX could significantly improve the results of therapy.Objective: to investigate the specific features of baseline (pretreatment) expression of genes responsible for major metabolic and energy production pathways in RA patients with different disease activity and to identify the genes, the baseline expression of which could serve as a predictor for remission attainment.Patients and methods. Blood from 40 RA patients (mean age 47.5 years; mean disease duration 7.9 weeks) who had not previously received MTX and 26 healthy donors (mean age 45.1 years). All the patients had used MTX (15 mg/week) for 2 years. Clinical response was evaluated by DAS28 and the serum levels of anti-cyclic citrullinated peptide antibodies, C-reactive protein, and rheumatoid factor. Remission was diagnosed according to ACR/EULAR and DAS28 (DAS28 <2.6). Joint structural changes were radiographically evaluated. Gene expression was determined in peripheral blood cells by real-time reverse transcriptase-polymerase chain reaction. A control group consisted of 26 randomly recruited gender- and sex-matched patients without autoimmune diseases and a family history.Results and discussion. MTX treatment significantly decreased disease activity according to DAS28. At the end of the investigation, the majority of patients had moderate disease activity (3.2≤ DAS28 ≤5.1), 4 had high disease activity, while 12 attained remission (DAS28 <2.6). Gene expression analysis showed that RA patients who had achieved clinical remission after MTX therapy displayed higher baseline expression of the genes associated with glycolysis (Glut1, PKM), inflammation (TNF-α), autophagy (ULK1), apoptosis (caspase 3, p21), and hypoxia (HIF1α), compared with patients who had not attained remission and with healthy individuals. In addition, in patients who had achieved remission, the baseline expression of the CD1 gene was significantly higher than in healthy individuals, while in the remaining patients the expression of this gene was significantly lower than in the controls. While the disease activity remained high, the baseline expression of the p21, caspase 3, TGFβ1, and RUNX2 genes was significantly lower than in healthy individuals and other patients with RA.Conclusion. Remission achievement in RA patients who had not previously received MTX was associated with higher baseline (pretreatment) gene expression associated with glycolytic activity, inflammation, autophagy, apoptosis, and hypoxia compared with patients who failed to attain remission. Elevated baseline expression of the CD1 gene compared with that in healthy individuals may serve as a predictor of sensitivity to MT therapy. 

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