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TSH-induced TNFα mRNA expression involves Akt and NF-κB. AKTi (A) or MG132 (B) was added 1 hour before TSH stimulation of healthy cultured fibrocytes. RNA was isolated after 6 hours of induction. AKTi treatment leads to significant reduction in TSH-induced TNFα mRNA expression (from 23-fold to 1.3-fold expression, p <0.0001) (A). Similarly, MG132 treatment inhibits TSH-induced TNFα mRNA expression (from 93-fold to 16-fold expression; p< 0.001) (B).
Source publication
Fibrocytes (FC) are bone marrow-derived progenitor cells that are more abundant and infiltrate the thyroid and orbit in Graves orbitopathy (GO). FCs express high levels of thyrotropin receptor (TSHR) and insulin-like growth factor-1 receptor (IGF-1R). These receptors are physically and functionally associated, but their role in GO pathogenesis is n...
Contexts in source publication
Context 1
... cultured fibrocytes, TSH-induced TNFα production is reduced by 52% and by 81% after AKTi and MG132 treatment, respectively (TSH-induced TNFα production of 1312 pg/ml is reduced to 612 pg/ml with AKTi and 251 pg/ml with MG132; p <0.0001; Fig 3B). AKTi treatment also leads to significant reduction in TSH- induced TNFα mRNA expression (from 23-fold to 1.3-fold expression, p <0.0001; Fig 4A). Similarly, MG132 treatment inhibits TSH-induced TNFα mRNA expression (from 93-fold to 16-fold expression p< 0.001; Fig 4B). ...
Context 2
... treatment also leads to significant reduction in TSH- induced TNFα mRNA expression (from 23-fold to 1.3-fold expression, p <0.0001; Fig 4A). Similarly, MG132 treatment inhibits TSH-induced TNFα mRNA expression (from 93-fold to 16-fold expression p< 0.001; Fig 4B). TSH/M22-induced TNFα production in fibrocytes is attenuated by TMB We investigated the impact of blocking IGF-1R signaling on TSH/M22-induced TNFα produc- tion in fibrocytes. ...
Similar publications
Teprotumumab, a monoclonal antibody targeted against the insulin-like growth factor-1 (IGF-1) receptor, was recently approved by the United States Food and Drug Administration (FDA) for the treatment of thyroid eye disease (TED). Phase I studies of teprotumumab for the treatment of malignancies demonstrated an acceptable safety profile but limited...
Citations
... Teprotumumab is a human monoclonal antibody against the IGF-1 receptor which, by binding to the receptor, inactivates it (5,25). It has been proven that teprotumumab reduces the TSH-induced release of pro-inflammatory cytokines by fibroblasts (45), and reduces the expression of the TSH receptor and IGF-1 receptor on the fibroblasts in patients with Graves'disease. The drug underwent phase II and III trials, showing significant effectiveness, especially in the reduction of proptosis and diplopia (5), and acceptable tolerance. ...
Graves' disease (GD) is the most common cause of hyperthyroidism in countries with adequate iodine supply, and its main extrathyroid manifestation is thyroid orbitopathy. For many years, thyrotoxicosis has been treated with antithyroid drugs, radioactive iodine therapy and thyroidectomy, and glucocorticoids have remained the standard treatment of thyroid eye disease (TED). Currently, there is a growing trend of using targeted therapies in the treatment of thyroid orbitopathy (TO). The significant development of immunobiology, a better understanding of the pathophysiology of the disease and a multitude of studies on new therapies give hope for better treatment results and an improvement in the quality of life of patients. The aim of the paper is to discuss the current principles of management of Graves' disease, including thyroid orbitopathy, with particular emphasis on new therapeutic methods. The potent role of K1-70TM and small molecule thyroid stimulating hormone (TSH) receptor antagonists in the treatment of hyperthyroidism is indicated. Not only do we point out the promising outcomes of combining glucocorticoids with mycophenolates, but also highlight the use of teprotumumab, rituximab, cyclosporin A, azathioprine, methotrexate, tocilizumab and sirolimus in the therapy of active moderate to severe form of orbitopathy. The protective and supportive effect of statins in the treatment of TED is noted. The limitations of the research results mentioned in the paper are remarked and the need for further randomized studies of long-term safety and efficacy in a larger group of patients is underlined.
... Il peut être donné par l'environnement du LT mais aussi par le fibrocyte. Les fibrocytes sont capables de sécréter de l'IL-12 et de l'IL-6 pouvant polariser les LT CD8 + vers un phénotype respectivement Tc1 et Tc17 (Chen et al., 2015a). ...
La bronchopneumopathie chronique obstructive (BPCO) est un problème de santé majeur, qui touche plus de 251 millions de personnes dans le monde. Le principal facteur de risque est l'exposition à la fumée de cigarette. La BPCO est caractérisée par une inflammation chronique des voies respiratoires et par un remodelage bronchique entraînant une diminution non réversible des débits d'air. Les patients atteints de BPCO sont souvent victimes d'épisodes d'exacerbations, qui sont des phases d'aggravation des symptômes respiratoires. Les traitements pharmacologiques actuels de la BPCO améliorent les symptômes et réduisent la fréquence des exacerbations mais n'ont aucun effet sur le déclin de la fonction respiratoire et la mortalité. La péribronche des patients atteints de BPCO est le siège d'une infiltration importante de cellules immunitaires et d'un remodelage tissulaire, permettant des contacts persistants entre cellules résidentes et cellules immunitaires. L'objectif de cette thèse est d'étudier si des contacts entre les cellules immunitaires et les cellules résidentes pourraient être impliqués dans la pathophysiologie de la BPCO. Plus particulièrement, j'ai cherché à savoir si les fibrocytes tissulaires, qui sont cellules produites par la moelle osseuse avec des propriétés fibroblastiques, peuvent interagir avec les lymphocytes T CD8+ (LT CD8+). De plus, je voulais savoir si le contact entre ces deux types de cellules pouvait être impliqué dans l'activation immunitaire chronique observable dans la BPCO. En utilisant des co-immunomarquages de spécimens bronchiques obtenus par chirurgie chez 17 patients atteints de BPCO et 25 sujets témoins, et des méthodes spécifiques d'analyse d'images, nous avons quantifié la distribution relative des fibrocytes et des LT CD8+. Des analyses transcriptomiques et des expériences fonctionnelles ont été utilisées pour étudier la capacité des LT CD8+ tissulaires à attirer les fibrocytes. Des co-cultures autologues directes et indirectes de fibrocytes et de LT CD8+, isolés à partir d'échantillons de sang de patients atteints de BPCO, ont été réalisées pour tester l'effet des fibrocytes sur la prolifération des LT CD8+ et le profil de sécrétion des cytokines. Nous avons défini un modèle mathématique, avec des règles et les paramètres dérivés de mes mesures expérimentales. Je montre que les fibrocytes et les LT CD8+ se trouvent à proximité dans les voies respiratoires distales et que les interactions (indirectes et directes) sont plus fréquentes dans les tissus des patients atteints de BPCO par rapport à ceux des sujets témoins. L'augmentation des interactions entre les LT CD8+ et les fibrocytes est associée à une altération de la fonction pulmonaire. J'ai démontré que les LT CD8+ tissulaires de patients atteints de BPCO favorisent le chimiotactisme des fibrocytes via l'axe CXCL2/8-CXCR1/2. Dans un essai in vitro, les LT CD8+ individuels établissent des interactions de courte durée avec les fibrocytes. Les contacts directs entre les deux types de cellules déclenchent la prolifération des LT CD8+ de manière dépendante de CD54 et CD86, ainsi que la production de cytokines pro-inflammatoires. La modélisation mathématique permet non seulement de décrire avec précision la distribution cellulaire selon que le sujet est sain ou atteint de BPCO, mais aussi de suivre la dynamique cellulaire. Au total, ma thèse révèle que des interactions locales entre fibrocytes et LT CD8+ peuvent se produire in vivo et pourraient compromettre l'équilibre entre immunité protectrice et inflammation chronique dans les bronches des patients atteints de BPCO.
... Teprotumumab, an insulin-like growth factor-1 receptor inhibitor, reduces the TSH-stimulated proinflammatory cytokines secretion of isolated fibrocytes [95] and significantly decreases the expression of TSH-R and IGF-1R in fibrocytes [96]. Teprotumumab significantly improved ptosis, diplopia, CAS score, and quality of life in moderate-to-severe GO and showed a favorable safety profile [97,98]. ...
Graves ophthalmopathy (GO), which occurs in autoimmune thyroid disease, can reduce patients’ quality of life due to its impact on visual function, physical appearance, and emotional health. Corticosteroids have been the first-line treatment for GO. More recently, the pathogenesis of GO has made significant progress. Various targeting biological agents and immunosuppressive agents make GO management more promising. Fully understanding GO pathogenesis and precise clinical management are beneficial for the prognosis of patients. Therefore, we conducted a comprehensive review of the medical management of GO and summarized research developments to highlight future research issues.
... Teprotumumab has been shown to decrease TSHR and IGF-1R display and reduce TSH/M22 stimulated cytokines and Akt phosphorylation in Graves' OFs and fibrocytes. [50,53] In two randomized, multicenter, double-masked, placebo-controlled, phase 2 and 3 clinical trials, published in 2017 and 2020, teprotumumab demonstrated a significant improvement in proptosis beginning at 6 weeks of treatment and over the course of 24 weeks compared to controls, with similar effects to surgical decompression in active moderate-to-severe TED patients. [3,54] At week 24, 83% in teprotumumab group had a reduction of proptosis ≥2 mm compared to 10% in the placebo group, and all secondary outcomes including overall response, CAS, diplopia, and quality of life score were significantly better in the teprotumumab group with minimal side effects. ...
Thyroid eye disease (TED) is the most common extrathyroidal manifestation of autoimmune Graves' hyperthyroidism. TED is a debilitating and potentially blinding disease with unclear pathogenesis. Autoreactive inflammatory reactions targeting orbital fibroblasts (OFs) lead to the expansion of orbital adipose tissues and extraocular muscle swelling within the fixed bony orbit. There are many recent advances in the understating of molecular pathogenesis of TED. The production of autoantibodies to cross-linked thyroid-stimulating hormone receptor and insulin-like growth factor-1 receptor (IGF-1R) activates OFs to produce significant cytokines and chemokines and hyaluronan production and to induce adipocyte differentiation. In moderately severe active TED patients, multicenter clinical trials showed that inhibition of IGF-1R with teprotumumab was unprecedentedly effective with minimal side effects. The emergence of novel biologics resulted in a paradigm shift in the treatment of TED. We here review the literature on advances of pathogenesis of TED and promising therapeutic targets and drugs.
... TSHR and IGF-1R not only formed a structural complex but also participated in crosstalk to mediate their function. Synergistic HA production was demonstrated by using IGF-1 combined with TSH (17), and IGF-1R inhibitors could block the signal transduction induced by TSH and GD-IgG (18,19). ...
Context
Thyroid-associated ophthalmopathy (TAO) is an organ-specific autoimmune disease closely associated with Graves’ disease (GD). IL-38, a novel cytokine in the IL-1 superfamily, has been reported to be involved in the pathogenesis of various autoimmune diseases.
Objectives
To evaluate the relationship between IL-38 and TAO disease activity and its role in inflammation and fibrosis in TAO.
Design/Setting/Participants
Blood samples and orbital connective tissues were collected from TAO patients and controls. Orbital fibroblasts (OFs) were isolated from patients with TAO.
Main Outcome
Measures Enzyme-linked immunosorbent assay (ELISA), immunohistochemistry (IHC), flow cytometry, immunofluorescence, quantitative real-time PCR and western blot analysis were performed.
Results
Here, we demonstrated that IL-38 levels decreased in the circulation and orbital connective tissues of patients with TAO compared to the controls, and were negatively correlated with the clinical activity score (CAS). In vitro, potent anti-inflammatory and anti-fibrotic effects of IL-38 were observed. Furthermore, we revealed that IL-38 can counteract the phosphorylation of star molecules in multiple classical pathways.
Conclusions
IL-38 plays a protective role in TAO and is associated with its pathogenesis. Our data suggest that IL-38 may be a promising marker of TAO disease activity and a potential target for TAO therapy.
... In cultured fibroblasts, this mAb suppressed the IGF-1/TSH-R-Ab-stimulated release of chemokines (190). Further, teprotumumab reduced the TSH-stimulated release of proinflammatory cytokines from isolated fibrocytes of GD patients (191), and markedly decreased the expression of both TSH-R and IGF-1R in fibrocytes (192). ...
Context
Invited update on the management of systemic autoimmune Graves disease (GD) and associated Graves orbitopathy (GO).
Evidence acquisition
Guidelines, pertinent original articles, systemic reviews, and meta-analyses.
Evidence synthesis
Thyrotropin receptor antibodies (TSH-R-Abs), foremost the stimulatory TSH-R-Abs, are a specific biomarker for GD. Their measurement assists in the differential diagnosis of hyperthyroidism and offers accurate and rapid diagnosis of GD. Thyroid ultrasound is a sensitive imaging tool for GD. Worldwide, thionamides are the favored treatment (12-18 months) of newly diagnosed GD, with methimazole (MMI) as the preferred drug. Patients with persistently high TSH-R-Abs and/or persistent hyperthyroidism at 18 months, or with a relapse after completing a course of MMI, can opt for a definitive therapy with radioactive iodine (RAI) or total thyroidectomy (TX). Continued long-term, low-dose MMI administration is a valuable and safe alternative. Patient choice, both at initial presentation of GD and at recurrence, should be emphasized. Propylthiouracil is preferred to MMI during the first trimester of pregnancy. TX is best performed by a high-volume thyroid surgeon. RAI should be avoided in GD patients with active GO, especially in smokers. Recently, a promising therapy with an anti-insulin-like growth factor-1 monoclonal antibody for patients with active/severe GO was approved by the Food and Drug Administration. COVID-19 infection is a risk factor for poorly controlled hyperthyroidism, which contributes to the infection–related mortality risk. If GO is not severe, systemic steroid treatment should be postponed during COVID-19 while local treatment and preventive measures are offered.
Conclusions
A clear trend towards serological diagnosis and medical treatment of GD has emerged.
... Anti-TNF-a TNF-a is a molecule produced by fibrocytes when they are stimulated by TSH or by thyroid-stimulating immunoglobulins. This molecule causes relevant effects, such as the production of adhesion molecules and chemokines in fibroblasts and the recruitment of inflammatory cells to local tissues [103]. Therefore, anti-TNF-a drugs could be useful despite their side effects [64,74], which include higher risk of infections, lupus-like reactions, immunogenicity, and demyelinating disorders [81,104]. ...
Introduction:
Thyroid eye disease (TED) is the most frequent extra-thyroid manifestation of Graves' disease and it is more frequent in middle age and in female gender. Nowadays, the causal mechanisms of this disease are not completely understood, but the current available studies suggest that the main causative factor is the thyroid stimulating hormone receptor.
Materials and methods:
To collect reports on TED medical management, a thorough literature search was performed in PubMed database. An additional search was made in Google Scholar to complete the collected items.
Results:
Among the indentified risk factors, tobacco habit is the most relevant. The main criteria to choose a suitable treatment are the activity and severity of the disease. Support measures can be used to improve the patient's symptoms in any phase of the disease. There is a large number of drugs proposed to manage TED, although with different reported rates of success.
Conclusions:
Currently, the drugs of choice are corticosteroids in moderate-to-severe and in sight-threatening forms. The main problem of corticosteroids is their spectrum of side effects. Therefore, other alternatives are being suggested for medical management of this disease. The efficacy of these alternatives remains unclear.
... Cytokine production by fibrocytes is stimulated by TSH and TSAbs, which interact with a receptor complex that is formed by the TSHR and the IGF-I receptor. Stimulation of this receptor complex results in activation of both the Akt and NF-κB pathways (51). As previously reported for thyroid cells, CD40 signaling can activate NF-κB on these orbit fibrocytes (52). ...
Nuclear factor-kappa B (NF-κB) is a ubiquitous transcription factor that is involved in inflammatory and immune responses, as well as in regulation of expression of many other genes related to cell survival, proliferation, and differentiation. In mammals, NF-κB comprises five subunits that can bind to promoter regions of target genes as homodimers or heterodimers. The most common dimer is the p50/p65 heterodimer. The several combinations of dimers that can be formed contribute to the heterogeneous regulation of NF-κB target genes, and this heterogeneity is further increased by interactions of the NF-κB dimers with other transcription factors, such as steroid hormone receptors, activator protein-1 (AP-1), and cAMP response element binding protein (CREB). In the thyroid, several studies have demonstrated the involvement of NF-κB in thyroid autoimmunity, thyroid cancer, and thyroid-specific gene regulation. The role of NF-κB in thyroid autoimmunity was hypothesized more than 20 years ago, after the finding that the binding of distinct NF-κB heterodimers to the major histocompatibility complex class I gene is hormonally regulated. Further studies have shown increased activity of NF-κB in thyroid autoimmune diseases and in thyroid orbitopathy. Increased activity of NF-κB has also been observed in thyroid cancer, where it correlates with a more aggressive pattern. Of particular interest, mutation of some oncogenes or tumor suppressor genes involved in thyroid carcinogenesis results in constitutive activation of the NF-κB pathway. More recently, it has been shown that NF-κB also has a role in thyroid physiology, as it is fundamental for the expression of the main thyroid-specific genes, such as sodium iodide symporter, thyroid peroxidase, thyroglobulin, Pax8, and TTF-1 (NKX2-1).
... TSH stimulation augmented the amount of cell-surface CD40, resulting in an increase in mean fluorescence intensity (MFI) from 1.96 ± 0.68 to 2.97 ± 0.98 (n = 4, P < 0.05) in healthy controls and from 1.88±0.63 to 3.23±0.78 in patients with GD (n = 4, P<0.05) ( Fig 1A). As we have observed previously for other proteins [17,18] TSHR signaling in fibrocytes from healthy donors and patients with GD displayed similar increases in cell-surface CD40 after TSH stimulation. Therefore, consecutive experiments were performed using fibrocytes from healthy donors. ...
Context:
Fibrocytes appear to participate in inflammation and tissue remodeling in patients with thyroid-associated ophthalmopathy (TAO). These patients have increased frequencies of circulating TSH receptor (TSHR)- and CD40-positive fibrocytes, suggesting TSHR and CD40 may play roles in proinflammatory cytokine production, which ultimately leads to orbital inflammation and tissue remodeling.
Objective:
To investigate the potential interactions between the TSHR and CD40 signaling pathways and their roles in IL-6 and TNF-α production.
Design and outcome measures:
CD40 expression on fibrocytes was assessed using flow cytometry; IL-6 and TNF-α protein release using Luminex technology; increased IL-6 and TNF-α mRNA abundance, using real-time PCR; TSH- and CD40 ligand (CD40L)-stimulated Akt phosphorylation in fibrocytes, by western blot analysis; TSHR-CD40 protein-protein interaction, using co-immunoprecipitation, and CD40-TSHR co-localization, using immunocytochemistry.
Results:
TSH enhances CD40 expression at a pre-translational level in fibrocytes. Production of IL-6 and TNF-α after costimulation with TSH and CD40L was greater than that after TSH or CD40L stimulation alone. TSH and CD40L costimulation also resulted in greater Akt phosphorylation. Akt and nuclear factor (NF)-κB inhibitors significantly reduced cytokine production after TSH and CD40L costimulation. TSHR and CD40L are colocalized on the cell surface and form a complex.
Conclusions:
TSHR and CD40 in fibrocytes appear to be physically and functionally related. TSH stimulates CD40 production on the fibrocyte surface. Cytokine expression upon simultaneous stimulation of TSHR and CD40 is greater than levels achieved with TSH or CD40L alone. Increased expression of CD40 by TSH is a potential mechanism for this process.
... Teprotumumab (RV 001, R1507) is a human monoclonal antibody that binds to the extracellular IGF-R1 domain designed for the treatment of hematologic and solid hematologic tumors [80]. Teprotumumab has been very recently demonstrated to reduce cell proliferation in GO fibrocytes [80] and it is currently used in a phase II randomized clinical trial in patients with active GO, to be completed in 2016 [81]. ...
C-X-C chemokine receptor (CXCR)3 and its interferon(IFN)γ-dependent chemokines (CXCL10, CXCL9, CXCL11) are implicated in the immune-pathogenesis of autoimmune thyroiditis (AT), Graves disease (GD) and Graves Ophthalmopathy (GO). In tissue, recruited Th1 lymphocytes produce IFNγ, enhancing the tissue secretion of IFNγ-inducible chemokines, initiating and perpetuating the autoimmune process. Patients with AT (with hypothyroidism), and with GO and GD, particularly in the active phase, have high IFNγ-inducible chemokines. Peroxisome proliferator-activated receptor (PPAR)γ or -α agonists and methimazole exert an immune-modulation on CXCR3 chemokines in AT, GD and GO. Other studies are ongoing to evaluate new molecules acting as antagonists of CXCR3, or blocking CXCL10, in Hashimoto thyroiditis (HT), GD and GO.
Recently, novel molecules targeting the various agents involved in the pathogenesis of GO, such as rituximab, have been proposed as an alternative to corticosteroids. However, randomized and controlled studies are needed to generalize these interesting results.
