Utilizing a PTPN22 gene signature to predict response to targeted therapies in rheumatoid arthritis

AbstractDespite the development of several targeted therapies for rheumatoid arthritis (RA), there is still no reliable drug-specific predictor to assist rheumatologists in selecting the most effective targeted therapy for each patient. Recently, a gene signature caused by impaired induction of PTPN22 in anti-CD3 stimulated peripheral blood mononuclear cells (PBMC) was observed in healthy at-risk individuals. However, the downstream target genes of PTPN22 and the molecular mechanisms regulating its expression are still poorly understood. Here we report that the PTPN22 gene signature is also present in PBMC from patients with active RA and can be reversed after effective treatment. The expression of PTPN22 correlates with that of more than 1000 genes in Th cells of anti-CD3 stimulated PBMC of healthy donors and is inhibited by TNFα or CD28 signals, but not IL-6, through distinct mechanisms. In addition, the impaired induction of PTPN22 in PBMC of patients with active RA can be normalized in vitro by several targeted therapies. More importantly, the in vitro normalization of PTPN22 expression correlates with clinical response to the targeted therapies in a longitudinal RA cohort. Thus, in vitro normalization of PTPN22 expression by targeted therapies can potentially be used to predict clinical response in a drug-specific manner.

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The molecular and cellular basis of corticosteroid resistance

Journal of Endocrinology ◽

10.1677/joe.0.1790301 ◽

2003 ◽

Vol 179 (3) ◽

pp. 301-310 ◽

Cited By ~ 20

Author(s):

IC Chikanza ◽

D Kozaci ◽

Y Chernajovsky

Keyword(s):

Rheumatoid Arthritis ◽

Peripheral Blood ◽

Molecular Mechanisms ◽

Mononuclear Cells ◽

Intracellular Signalling ◽

Signalling Pathways ◽

Peripheral Blood Mononuclear ◽

Intracellular Signalling Pathways ◽

Nf Kappab

Corticosteroids (CS) can modulate gene expression and are often used to treat a range of immunological and inflammatory diseases such as asthma, inflammatory bowel disease and rheumatoid arthritis. However, a proportion of patients fail to show an adequate response. On this basis patients have been subdivided into CS-sensitive (SS) and -resistant (SR) subgroups. The ability of CS to inhibit peripheral blood T cell proliferation in vitro has also been used similarly. In rheumatoid arthritis (RA), the in vitro-defined SS and SR subgroups correlate with the clinical responses to CS therapy. The mechanisms responsible for this observation are unknown but they appear to involve a number of known molecular events related to the described mechanisms of action of CS. These include alterations in the functional status of CS receptor-alpha, perturbations of the cytokine and hormonal milieu and intracellular signalling pathways. Peripheral blood mononuclear cells (MNCs) from SR significantly overexpress activated NF-kappaB. In vitro, CS fail to significantly inhibit concanavalin A (conA)-induced NF-kappaB activation in MNCs from SR RA patients. The alterations in the intracellular signalling pathways may explain in part our observations seen in SR RA subjects, CS fail to significantly inhibit conA-induced interleukin (IL)-2 and IL-4 secretion and lipopolysaccharide-induced IL-8 and IL-1beta secretion in vitro. CS therapy fails to reduce the circulating levels of IL-8 and IL-1beta in RA patients. In asthma, CS fail to induce L10 in SR asthma patients. Other molecular mechanisms such as enhanced AP-1 expression and alterations in the MAP kinase pathway are most likely to be involved too and we are currently investigating such possibilities. A full understanding of the molecular basis of SR will lead to the development of more rational therapeutic strategies.

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Effect in vitro of a bacterial extract (OM-89) on interleukin 1 and interleukin 2 production by peripheral blood mononuclear cells from healthy subjects and rheumatoid arthritis patients

International Journal of Immunopharmacology ◽

10.1016/0192-0561(90)90011-b ◽

1990 ◽

Vol 12 (8) ◽

pp. 909-913 ◽

Cited By ~ 3

Author(s):

Jacques Clot ◽

Monique Andary

Keyword(s):

Rheumatoid Arthritis ◽

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Healthy Subjects ◽

Mononuclear Cells ◽

Interleukin 1 ◽

Interleukin 2 ◽

Peripheral Blood Mononuclear ◽

Blood Mononuclear Cells

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Utilizing a PTPN22 gene signature to predict response to targeted therapies in rheumatoid arthritis

Journal of Autoimmunity ◽

10.1016/j.jaut.2019.04.012 ◽

2019 ◽

Vol 101 ◽

pp. 121-130 ◽

Cited By ~ 2

Author(s):

Hui-Hsin Chang ◽

Ching-Huang Ho ◽

Beverly Tomita ◽

Andrea A. Silva ◽

Jeffrey A. Sparks ◽

...

Keyword(s):

Rheumatoid Arthritis ◽

Targeted Therapies ◽

Gene Signature ◽

Ptpn22 Gene

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Glucocorticoids and Catecholamines Affect in Vitro Functionality of Porcine Blood Immune Cells

Animals ◽

10.3390/ani9080545 ◽

2019 ◽

Vol 9 (8) ◽

pp. 545 ◽

Cited By ~ 3

Author(s):

Lena Reiske ◽

Sonja Schmucker ◽

Julia Steuber ◽

Volker Stefanski

Keyword(s):

Immune Cells ◽

Lymphocyte Proliferation ◽

Molecular Mechanisms ◽

Mononuclear Cells ◽

Intracellular Cytokine Staining ◽

Stress Hormones ◽

High Stress ◽

Peripheral Blood Mononuclear ◽

Indwelling Catheters

Stress hormones exert important modulating influences on the functionality of immune cells. Despite its major role as a livestock animal and its increasing use as an animal model, knowledge about this relationship in the domestic pig is rare. This study therefore aimed to characterize the effect of glucocorticoids and catecholamines on the proliferation and cytokine production of porcine peripheral blood mononuclear cells (PBMC). Blood was obtained from donor pigs equipped with indwelling catheters to exclude stress hormone exposition before in vitro testing. PBMC were stimulated in the presence of cortisol, adrenaline or noradrenaline at concentrations resembling low to high stress conditions. Proliferation was determined via 3H-thymidine incorporation, and TNFα producers were quantified by intracellular cytokine staining. Cortisol led to a decrease in mitogen-induced lymphocyte proliferation and the number of TNFα producing cells. In contrast, catecholamines increased proliferation while exerting repressive or no effects on the number of cytokine producers. Remarkably, in concentrations presumably found in lymphatic tissue in stress situations, noradrenaline suppressed lymphocyte proliferation completely. The shown repressive effects might especially have implications on health and welfare in pigs. The obtained results provide a preliminary database for extended studies on the molecular mechanisms of glucocorticoid and catecholamine actions on porcine immune cells.

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In vitro effects of Epstein-Barr virus on peripheral blood mononuclear cells from patients with rheumatoid arthritis and normal subjects.

Journal of Experimental Medicine ◽

10.1084/jem.148.5.1429 ◽

1978 ◽

Vol 148 (5) ◽

pp. 1429-1434 ◽

Cited By ~ 165

Author(s):

L Slaughter ◽

D A Carson ◽

F C Jensen ◽

T L Holbrook ◽

J H Vaughan

Keyword(s):

Rheumatoid Arthritis ◽

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Epstein Barr Virus ◽

Mononuclear Cells ◽

Peripheral Blood Mononuclear ◽

Barr Virus ◽

Blood Mononuclear Cells ◽

Epstein Barr

Peripheral blood mononuclear cells from 10 patients with rheumatoid arthritis and 9 control subjects were cultured in vitro for 30 days with and without infection by Epstein-Barr virus. All cultures showed polyclonal stimulation of B cells as indicated by rising levels of IgM in the culture supernates, reaching maximal at 18-24 days, and with no quantitative or kinetic difference between the RA and control cells. IgM anti-IgG was also produced in both groups and maximally at 18-24 days, but in greater quantity by the RA lymphocytes. The anti-IgG made by the RA lymphocytes was more easily absorbed by solid phase IgG than was the anti-IgG made by the normal lymphocytes and thus was judged to be of higher affinity. RA lymphocytes uninfected with EBV had higher transformation scores than did the normal controls and developed spontaneously into permanent cell lines in six instances.

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Preclinical Antileukemia Activity of Tramesan: A Newly Identified Bioactive Fungal Metabolite

Oxidative Medicine and Cellular Longevity ◽

10.1155/2017/5061639 ◽

2017 ◽

Vol 2017 ◽

pp. 1-8 ◽

Cited By ~ 5

Author(s):

M. R. Ricciardi ◽

R. Licchetta ◽

S. Mirabilii ◽

M. Scarpari ◽

A. Parroni ◽

...

Keyword(s):

Cell Lines ◽

Molecular Mechanisms ◽

Mononuclear Cells ◽

Leukemic Cell ◽

Bioactive Compound ◽

Primary Cells ◽

Peripheral Blood Mononuclear ◽

Normal Peripheral Blood ◽

Leukemic Cell Lines

Despite improvements that occurred in the last decades in the acute myeloid leukemia (AML) treatment, clinical results are still unsatisfactory. More effective therapies are required, and innovative approaches are ongoing, including the discovery of novel antileukemia natural compounds. Several studies have described the activity of extracts from mushrooms which produce compounds that exhibited immunological and antitumor activities. The latter has been demonstrated to be promoted in vitro by mushroom polysaccharides via induction of apoptosis. However, the antileukemia activity of these compounds on primary cells is still not reported. In the present study, we examined the in vitro effects of Tramesan (TR), a bioactive compound extracted from Trametes versicolor, on leukemic cell lines and primary cells. Our results demonstrated that TR induced a marked growth inhibition of leukemic cell lines and primary cells from AML patients. The antiproliferative effects of TR were associated in primary AML cells with a significant increase of apoptosis. No significant cytotoxic effects were observed in normal peripheral blood mononuclear cells (MNC) from healthy donors. Our data demonstrated a cytotoxic activity of TR on leukemia cells prompting further translational applications. Ongoing studies are elucidating the molecular mechanisms underlying its antileukemic activity.

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Decreased miR-4512 Levels in Monocytes and Macrophages of Individuals With Systemic Lupus Erythematosus Contribute to Innate Immune Activation and Neutrsophil NETosis by Targeting TLR4 and CXCL2

Frontiers in Immunology ◽

10.3389/fimmu.2021.756825 ◽

2021 ◽

Vol 12 ◽

Author(s):

Binbin Yang ◽

Xinwei Huang ◽

Shuangyan Xu ◽

Li Li ◽

Wei Wu ◽

...

Keyword(s):

Systemic Lupus Erythematosus ◽

Lupus Erythematosus ◽

Target Genes ◽

Mononuclear Cells ◽

Therapeutic Potential ◽

Luciferase Reporter ◽

Systemic Lupus ◽

Peripheral Blood Mononuclear ◽

Monocytes And Macrophages

ObjectiveSystemic lupus erythematosus (SLE) is an autoimmune disease with complex etiology that is not yet entirely understood. We aimed to elucidate the mechanisms and therapeutic potential of microRNAs (miRNAs) in SLE in a Tibetan population.MethodsPeripheral blood mononuclear cells from SLE patients (n = 5) and healthy controls (n = 5) were used for miRNA–mRNA co-sequencing to detect miRNAs related to immune abnormalities associated with SLE. Luciferase reporter assay was used to identify potential targets of candidate miRNA. The target genes were verified in miRNA-agomir/antagomir transfection assays with multiple cells lines and by expression analysis. The effects of candidate miRNA on monocyte and macrophage activation were evaluated by multiple cytokine profiling. Neutrophil extracellular traps (NETs) formation was analyzed in vitro by cell stimulation with supernatants of monocytes and macrophages transfected with candidate miRNA. The rodent MRL/lpr lupus model was used to evaluate the therapeutic effect of CXCL2Ab on SLE and the regulation effect of immune disorders.ResultsIntegrated miRNA and mRNA expression profiling identified miRNA-4512 as a candidate miRNA involved in the regulation of neutrophil activation and chemokine-related pathways. MiR-4512 expression was significantly reduced in monocytes and macrophages from SLE patients. MiR-4512 suppressed the TLR4 pathway by targeting TLR4 and CXCL2. Decreased monocyte and macrophage miR-4512 levels led to the expression of multiple proinflammatory cytokines in vitro. Supernatants of miR-4512 antagomir-transfected monocytes and macrophages significantly promoted NETs formation (P < 0.05). Blocking of CXCL2 alleviated various pathogenic manifestations in MRL/lpr mice, including kidney damage and expression of immunological markers of SLE.ConclusionsWe here demonstrated the role of miR-4512 in innate immunity regulation in SLE. The effect of miR-4512 involves the regulation of monocytes, macrophages, and NETs formation by direct targeting of TLR4 and CXCL2, indicating the miR-4512-TLR4-CXCL2 axis as a potential novel therapeutic target in SLE.

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PhosSNPs-Regulated Gene Network and Pathway Significant for Rheumatoid Arthritis

Human Heredity ◽

10.1159/000518608 ◽

2021 ◽

pp. 1-11

Author(s):

Pei He ◽

Fei Jiang ◽

Wei Guo ◽

Yu-Fan Guo ◽

Shu-Feng Lei ◽

...

Keyword(s):

Rheumatoid Arthritis ◽

Oxidative Phosphorylation ◽

Protein Phosphorylation ◽

Gene Network ◽

Target Genes ◽

Mononuclear Cells ◽

Peripheral Blood Mononuclear ◽

Chinese Sample ◽

Oxidative Phosphorylation Pathway ◽

Public Datasets

Objectives: Peripheral blood mononuclear cells (PBMCs) are critical for immunity and participate in multiple human diseases, including rheumatoid arthritis (RA). PhosSNPs are nonsynonymous SNPs influencing protein phosphorylation, thus probably modulate cell signaling and gene expression. We aimed to identify phosSNPs-regulated gene network/pathway potentially significant for RA. Methods: We collected genome-wide phosSNP genotyping data and transcriptome-wide mRNA expression data from PBMCs of a Chinese sample. We discovered and verified with public datasets differentially expressed genes (DEGs) associated with RA, and replicated RA-associated SNPs in our study sample. We performed a targeted expression quantitative trait locus (eQTL) study on significant phosSNPs and DEGs. Results: We identified 29 nominally significant eQTL phosSNPs and 83 target genes, and constructed comprehensive regulatory/interaction networks, highlighting the vital effects of two eQTL phosSNPs (rs371513 and rs4824675, FDR <0.05) and four critical node genes (HSPA4, NDUFA2, MRPL15, and ATP5O). Besides, two node/key genes NDUFA2 and ATP5O, regulated by rs371513, were significantly enriched in mitochondrial oxidative phosphorylation pathway. Besides, four pairs of eQTL effects were replicated independently in whole blood and/or transformed fibroblasts. Conclusions: The findings delineated a potential role of protein phosphorylation and genetic variations in RA and warranted the significant roles of phosSNPs in regulating RA-associated genes expression in PBMCs. The results pointed out the relevance and significance of oxidative phosphorylation pathway to RA.

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