scholarly journals OP0245 ANTI-S100A4 MONOCLONAL ANTIBODY TREATMENT AMELIORATES SKIN FIBROSIS IN INFLAMMATORY AND NON-INFLAMMATORY PRE-CLINICAL MODELS OF SYSTEMIC SCLEROSIS

2021 ◽  
Vol 80 (Suppl 1) ◽  
pp. 150.1-150
Author(s):  
M. Tomčík ◽  
T. Trinh-Minh ◽  
C. T. Manh ◽  
H. Štorkánová ◽  
L. Štorkánová ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Systemic Sclerosis ◽  
Skin Fibrosis ◽  
Hydroxyproline Content ◽  
Control Groups ◽  
Antibody Treatment ◽  
Fibroblast Activation ◽  
Dermal Thickness ◽  
Dosing Regimens ◽  
Clinical Models

Background:AX-202 is a monoclonal antibody that inhibits the bioactivity of S100A4. S100A4 is an alarm signal that is released from cells in response to stress or injury and functions as an amplifying mechanism of inflammation and fibrosis in the diseased tissue microenvironment. Previous in vitro studies have found that S100A4 induces fibroblast activation, sensitizes fibroblasts to the effects of TGFβ, drives epithelial-mesenchymal transition, and stimulates monocyte cytokine release (1-3). Moreover, S100A4-/- mice are protected from fibrosis in several animal models (1). In patients with systemic sclerosis (SSc), S100A4 is elevated both in lesional tissue and systemically and correlates with skin involvement, disease activity, and pulmonary function.Objectives:The aim of this study was to assess the antifibrotic effects of murine AX-202 in two pre-clinical models of SSs reflecting both inflammation-mediated and inflammation non-mediated fibrosis and confirm the in vivo activity of humanized AX-202.Methods:We first evaluated the effects of murine AX-202 in the bleomycin-induced skin fibrosis model and the tight-skin 1 (Tsk-1) model. In the bleomycin (BLM) model, fibrosis was induced by 3 weeks of BLM s.c. injections followed by 3 weeks of AX-202 treatment in parallel with continued BLM s.c. injections. The control groups included NaCl s.c. injections for 6 weeks, BLM s.c. injections for 6 weeks, or BLM s.c. injections for 3 weeks, followed by NaCl s.c. injections for 3 weeks. Three dosing regimens of AX-202 were tested: 3.75, 7.5, or 12.5 mg/kg i.p. every 3rd day. In the Tsk-1 model, treatment with 7.5 mg/kg i.p. every 3rd day was administered from week 5 until week 10. The control groups included pa mice, Tsk-1 mice, and Tsk-1 mice treated i.p. with isotype IgG. We subsequently evaluated the effects of humanized AX-202 in the model of BLM-induced skin fibrosis in a similar design as used for the murine AX-202 study. Three dosing regimens were tested: 8 mg/kg and 16 mg/kg i.p. every 3rd day and 24 mg/kg i.v. once weekly.Results:In the BLM model, murine AX-202 (7.5 mg/kg) was effective both in the prevention of progression of pre-established skin fibrosis and in the induction of regression of fibrosis as assessed by the dermal thickness (-55%, p<0.0001 vs BLM for 6 weeks, and -23%, p<0.0001 vs BLM for 3 weeks), myofibroblast count and hydroxyproline content. Murine AX-202 also ameliorated fibrosis in the Tsk-1 model as assessed by the hypodermal thickness (-24%, p=0.01 vs Tsk-1 isotype control), myofibroblast count, and hydroxyproline content. In both models, the antifibrotic effects were associated with a reduction in pSMAD3 expression. Humanized AX-202 was effective in the prevention of progression of pre-established skin fibrosis in all doses tested across all endpoints (dermal thickness, myofibroblast counts, hydroxyproline content). In the two groups treated with 16 mg/kg i.p. and 24 mg/kg i.v., humanized AX-202 also induced regression of fibrosis (-83%, p<0.001, and -61%, p<0.001 vs BLM for 3 weeks, respectively). Both murine and humanized AX-202 were well tolerated in all study groups in both models.Conclusion:We demonstrate that AX-202 confers potent antifibrotic effects in complementary models of SSc. These results confirm and expand previous data showing that inhibition of S100A4 by AX-202 is a promising potential therapeutic candidate for disease modification in SSc or other fibrotic conditions.References:[1]Tomcik M et al. S100A4 amplifies TGF-beta-induced fibroblast activation in systemic sclerosis. Ann Rheum Dis. 2015;74(9):1748-55.[2]Cerezo LA et al. The metastasis-associated protein S100A4 promotes the inflammatory response of mononuclear cells via the TLR4 signalling pathway in rheumatoid arthritis. Rheumatology (Oxford). 2014;53(8):1520-6.[3]Fei F, et al. Role of metastasis-induced protein S100A4 in human non-tumor pathophysiologies. Cell Biosci. 2017;7:64.Acknowledgements:The study was supported by Arxx Therapeutics and MHCR 023728.Disclosure of Interests:Michal Tomčík: None declared, Thuong Trinh-Minh: None declared, Cuong Tran Manh: None declared, Hana Štorkánová: None declared, Lenka Štorkánová: None declared, Ladislav Šenolt: None declared, Jörg Klingelhöfer Employee of: Arxx Therapeutics, Rizwan I Hussain Employee of: Arxx Therapeutics, Jonas Hallén Employee of: Arxx Therapeutics, Jörg H.W. Distler Shareholder of: the stock owner of 4D Science, Consultant of: Actelion, Active Biotech, Anamar, ARXX, Bayer Pharma, Boehringer Ingelheim, Celgene, Galapagos, GSK, Inventiva, JB Therapeutics, Medac, Pfizer, RuiYi and UCB, Grant/research support from: Anamar, Active Biotech, Array Biopharma, ARXX, aTyr, BMS, Bayer Pharma, Boehringer Ingelheim, Celgene, Galapagos, GSK, Inventiva, Novartis, Sanofi-Aventis, RedX, UCB

scholarly journals Rituximab in systemic sclerosis

2021 ◽  
Vol 30 (2) ◽  
pp. 31-35
Author(s):  
T. Shevtsova ◽  
V. Nadtocheeva ◽  
S. Nimiritskaya ◽  
L. Akulkina ◽  
N. Bulanov ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Systemic Sclerosis ◽  
B Cells ◽  
Interstitial Lung Disease ◽  
Lung Disease ◽  
Skin Fibrosis

To evaluate the effect of intravenous rituximab, a monoclonal antibody to B-cells, on interstitial lung disease, skin fibrosis and arthritis in patients with systemic sclerosis (SSc).

Preventive Role of Vitamin D in Silica-Induced Skin Fibrosis: A Study in Relation to Oxidative Stress and Pro-Inflammatory Cytokines

2016 ◽  
Vol 86 (3-4) ◽  
pp. 88-96 ◽  
Author(s):  
Ashwani Koul ◽  
Stanzin Angmo ◽  
Sanjay Bharati
Keyword(s):  
Oxidative Stress ◽  
Lipid Peroxidation ◽  
Vitamin D ◽  
Intradermal Injection ◽  
Skin Fibrosis ◽  
Protective Effects ◽  
Hydroxyproline Content ◽  
Dermal Thickness ◽  
Tnf Α ◽  
Antioxidant Defence System

Abstract.The protective effects of vitamin D analogue calcipotriol in silica-induced skin fibrosis were studied in the present study. Male BALB / c mice were divided into four groups; Control, Vitamin D, Silica and Silica+Vitamin D. Silica was administered as a single intradermal injection (40 µg / µL, dissolved in normal saline; particle size 1 – 5 µm) in the hind limbs of animals in Silica & Silica+Vitamin D group. Vitamin D group animals received topical application of 100µL of vitamin D solution (10-7M in Ethanol) daily for 12 weeks. Silica+Vitamin D group animals received co-treatment of silica and vitamin D as described for other groups. After 12 weeks of treatment, dermal thickness and hydroxyproline content of treated sections were measured. The TNF-α and IL-6 levels were measured in serum of all treated animals. The silica-induced oxidative stress was measured in terms of lipid peroxidation in skin tissue. Antioxidant defence system was assessed in terms of levels of reduced glutathione, superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase. A significant increase in the dermal thickness and hydroxyproline content was observed after silica treatment (931 ± 57.98 to 1804.61 ± 146.20 µm)(p < 0.05). Vitamin D co-treatment reduced dermal thickness and hydroxyproline content compared to Silica group (p < 0.05). Similarly a decrease in TNF-α and IL-6 levels were also observed after vitamin D treatment. A significant reduction in oxidative stress in terms of lipid peroxidation (4.92 ± 0.70 to 2.40 ± 0.31 nmol / mg protein). Therefore, present study suggested that vitamin D could be an effective agent against silica-induced skin fibrosis and oxidative stress.

scholarly journals SAT0291 THE ROLE OF X-LINKED INHIBITOR OF APOPTOSIS PROTEIN (XIAP) IN SYSTEMIC SCLEROSIS

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 1090.1-1090
Author(s):  
C. Bergmann ◽  
L. Hallenberger ◽  
B. Merlevede ◽  
C. Dees ◽  
C. W. Chen ◽  
...  
Keyword(s):  
Systemic Sclerosis ◽  
Target Genes ◽  
Skin Fibrosis ◽  
Inhibitor Of Apoptosis ◽  
Research Support ◽  
Inhibitor Of Apoptosis Protein ◽  
Fibroblast Activation ◽  
Apoptosis Protein

Background:Pathologic activation of fibroblasts is a central feature of fibrotic tissue disease in Systemic Sclerosis (SSc). Although individual key signaling pathways of fibroblast activation such as transforming growth factor β (TGFβ) and WNT/β-catenin signaling have been identified, the consequences of the concomitant upregulation of these pathways and their crosstalk are incompletely characterized. Given the high medical need, the identification of mutual activation and amplification loops of profibrotic signals is essential to identify novel candidates for antifibrotic therapies. XIAP (X-linked inhibitor of apoptosis protein) is a ubiquitously expressed member of the IAP protein family which are implicated in the regulation of various cellular functions and tissue turnover. XIAP was recently described to be implicated in WNT/β-catenin signaling and TGFβ signaling.Objectives:The aim of this study is to characterize the role of XIAP in fibrotic disease.Methods:XIAP-expression was analyzed by qPCR, IF and Western blot. XIAP was targeted pharmacologically and with siRNA. The activation of WNT/β-catenin signaling was assessed by analyses of WNT target genes, by TOPflash/FOPflash luciferase reporter assay and in reporter mice.In vivo,XIAP inhibition was analysed in two different models of fibrosis.Results:The expression of XIAP is increased in the skin of SSc patients compared to matched healthy individuals with a particular prominent expression in fibroblasts. The overexpression of XIAP is more pronounced in SSc patients with diffuse and active skin fibrosis compared to SSc patients with limited and inactive disease. The overexpression of XIAP is also reflected in several experimental fibrosis models: the model of sclerodermatous graft versus host disease, the model of bleomycin induced skin fibrosis and Topoisomerase I induced fibrosis (TopoI) mice. TGFβ induces the expression of XIAP in vitro and in vivo and treatment with the TGFβ1 receptor antagonist SD208 reverses the TGFβ induced expression of XIAP. Inhibition of XIAP with embelin or siRNA reduces the TGFβ induced activation of fibroblasts with reduced collagen release and reduced expression of myofibroblast markers. In addition, XIAP inhibition reverted the activated fibroblast phenotype in SSc fibroblasts with reduced expression of stress fibers and αSMA. The antifibrotic effects of XIAP inhibition occurred in non-toxic doses as demonstrated by MTT and by TUNEL staining. In vivo, inhibition of XIAP reduced skin fibrosis in the models of bleomycin induced skin fibrosis and in TopoI-induced skin and lung fibrosis as demonstrated by analysis of dermal thickening, dermal hydroxyproline content and by analysis of myofibroblast differentiation. Mechanistically, XIAP inhibition reduced the activation of WNT/β-catenin signaling as demonstrated by TOPflash reporter assays and by the analysis of WNT target genes.Conclusion:XIAP is upregulated in SSc fibroblasts and murine SSc models in a TGFβ-dependent manner and promotes fibroblast activation by fostering canonical WNT signaling. Our data suggest that XIAP mediates an amplification loop between TGFβ and WNT/β-catenin signaling. Inhibition of XIAP may thus be a novel approach to target aberrant WNT/β-catenin signaling in fibrotic diseases.Disclosure of Interests:Christina Bergmann: None declared, Ludwig Hallenberger: None declared, Benita Merlevede: None declared, Clara Dees: None declared, Chih-Wei Chen: None declared, Oliver Distler Grant/research support from: Grants/Research support from Actelion, Bayer, Boehringer Ingelheim, Competitive Drug Development International Ltd. and Mitsubishi Tanabe; he also holds the issued Patent on mir-29 for the treatment of systemic sclerosis (US8247389, EP2331143)., Consultant of: Consultancy fees from Actelion, Acceleron Pharma, AnaMar, Bayer, Baecon Discovery, Blade Therapeutics, Boehringer, CSL Behring, Catenion, ChemomAb, Curzion Pharmaceuticals, Ergonex, Galapagos NV, GSK, Glenmark Pharmaceuticals, Inventiva, Italfarmaco, iQvia, medac, Medscape, Mitsubishi Tanabe Pharma, MSD, Roche, Sanofi and UCB, Speakers bureau: Speaker fees from Actelion, Bayer, Boehringer Ingelheim, Medscape, Pfizer and Roche, Georg Schett Speakers bureau: AbbVie, BMS, Celgene, Janssen, Eli Lilly, Novartis, Roche and UCB, Jörg Distler Grant/research support from: Boehringer Ingelheim, Consultant of: Boehringer Ingelheim, Paid instructor for: Boehringer Ingelheim, Speakers bureau: Boehringer Ingelheim

scholarly journals AB0091 INHIBITION OF AUTOPHAGY PREVENTS PROGRESSION OF FIBROSIS IN MURINE MODELS OF SYSTEMIC SCLEROSIS

2021 ◽  
Vol 80 (Suppl 1) ◽  
pp. 1075.3-1075
Author(s):  
A. Zehender ◽  
Y. N. Li ◽  
N. Y. Lin ◽  
A. H. Györfi ◽  
A. Soare ◽  
...  
Keyword(s):  
Systemic Sclerosis ◽  
Molecular Mechanisms ◽  
Interstitial Fibrosis ◽  
Human Fibroblasts ◽  
Skin Fibrosis ◽  
Autophagic Flux ◽  
Murine Models ◽  
Tissue Fibrosis ◽  
Dermal Thickness

Background:Autophagy is catabolic process allowing cells to degrade unnecessary or dysfunctional cellular organelles. Failure of appropriate regulation of autophagy, however, can severely perturb tissue homeostasis. Recent studies demonstrate that autophagy is activated in several fibrotic diseases such as liver fibrosis, renal interstitial fibrosis, cardiac fibrosis.Objectives:The objective of this work was to characterize the activation of autophagy in systemic sclerosis (SSc) and to decipher its role in the pathogenesis of SSc.Methods:Activation of autophagy in skin samples of patients and murine models of SSc was assessed by co-staining of LC3B and P62 with the lysosomal marker LAMP2. The role of the autophagy was investigated in the model of bleomycin-induced dermal fibrosis. Beclin1 was overexpressed using adenovirus encoding for Beclin1. To knockdown Atg7 in vivo was achieved by subcutaneous injections of Atg7 siRNA or non-targeting siRNA. In vivo, 3-methyladenine (3-MA) was administered i.p. in a concentration of 15 mg/kg ones daily. Protein expression was measured by Western blot. Target genes were analyzed by qPCR. To monitor the autophagic flux, we generated adenoviral vectors encoding for tandem fluorescent-tagged LC3 (mRFP-EGFP-LC3).Results:In the present study, we demonstrate that autophagy is activated in fibroblasts in SSc skin and also in experimental fibrosis models as compared to respective non-fibrotic control tissue with enhanced activity in in vivo and in vitro autophagy reporter studies. The aberrant activation of autophagy had profound stimulatory effects on fibroblasts. Activation of autophagy by forced expression of BECLIN1 promoted fibroblast-to-myofibroblast transition and stimulated the collagen release by cultured human fibroblasts and induced fibrosis in murine model. Nevertheless, inhibition of autophagy can deactivate myofibroblasts and induce regression of tissue fibrosis. Knockdown of ATG7 or BECLIN1 in human fibroblasts reduced the expression of αSMA and the number of stress fibers in myofibroblasts, indicating re-differentiation of myofibroblasts into resting fibroblasts upon inhibition of autophagy. Similar results were obtained with the autophagy inhibitors CQ and 3-MA. In vivo, siRNA mediated knockdown of Atg7 effectively prevented progression of fibrosis in a model of established bleomycin-induced skin fibrosis. Inactivation of autophagy decreased dermal thickness, myofibroblast counts and hydroxyproline content to below pretreatment levels, indicating regression of bleomycin-induced skin fibrosis. In addition, treatment of mice with the autophagy inhibitor 3-MA ameliorated bleomycin-induced skin fibrosis.Conclusion:We demonstrate that autophagy activity is enhanced in fibroblasts of SSc patients and in murine models of SSc. The increased activation of autophagy induces fibroblast-to-myofibroblast transition and promotes fibrotic tissue remodeling. However, inhibition of autophagy can deactivate myofibroblasts and induce regression of tissue fibrosis.References:[1]Wynn, T. Cellular and molecular mechanisms of fibrosis. J Pathol 214, 199-210 (2008).[2]Klionsky DJ, Abeliovich H, Agostinis P, et al. Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes. Autophagy 4, 151-175 (2008).[3]Wang, CW & Klionsky, DJ. The molecular mechanism of autophagy. Mol Med 9, 65-76 (2003).[4]Hernández-Gea V, Ghiassi-Nejad Z, Rozenfeld R, et al. Autophagy releases lipid that promotes fibrogenesis by activated hepatic stellate cells in mice and in human tissues. Gastroenterology 142, 938-946 (2012).Disclosure of Interests:Ariella Zehender: None declared, Yi-Nan Li: None declared, Neng-Yu Lin: None declared, Andrea-Hermina Györfi: None declared, Alina Soare: None declared, Christina Bergmann: None declared, Andreas Ramming: None declared, Georg Schett: None declared, Jörg H.W. Distler Consultant of: Actelion, Active Biotech, Anamar, ARXX, Bayer Pharma, Boehringer Ingelheim, Celgene, Galapagos, GSK, Inventiva, JB Therapeutics, Medac, Pfizer, RuiYi and UCB., Grant/research support from: Anamar, Active Biotech, Array Biopharma, aTyr, BMS, Bayer Pharma, Boehringer Ingel-heim, Celgene, Galapagos, GSK, Inventiva, Novartis, Sanofi-Aventis, RedX, UCB., Employee of: stock owner of 4D Science and Scientific head of FibroCure

scholarly journals Src kinases in systemic sclerosis: Central roles in fibroblast activation and in skin fibrosis

2008 ◽  
Vol 58 (5) ◽  
pp. 1475-1484 ◽  
Author(s):  
Catherine Skhirtladze ◽  
Oliver Distler ◽  
Clara Dees ◽  
Alfiya Akhmetshina ◽  
Nicole Busch ◽  
...  
Keyword(s):  
Systemic Sclerosis ◽  
Skin Fibrosis ◽  
Src Kinases ◽  
Fibroblast Activation

Inhibition of IL-13: A New Pathway for Atopic Dermatitis

2020 ◽  
pp. 120347542098255
Author(s):  
Kayadri Ratnarajah ◽  
Michelle Le ◽  
Anastasiya Muntyanu ◽  
Steve Mathieu ◽  
Simon Nigen ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Atopic Dermatitis ◽  
Severity Index ◽  
Severe Atopic Dermatitis ◽  
Treatment Alternative ◽  
Control Groups ◽  
Rapid Improvement ◽  
Common Receptor ◽  
The Common ◽  
And Control

Dupilumab, a monoclonal antibody against the common receptor of interleukin (IL)-4 and IL-13, was the first biologic therapy approved in Canada for treatment of moderate-to-severe atopic dermatitis (AD). While it is considered safe and effective, dupilumab is not universally effective and 8%-38% of patients develop conjunctivitis, while some patients develop head and neck dermatitis. Thus, new therapeutic options are warranted. While both IL-4 and IL-13 play important roles in the pathogenesis of AD, it has been recently demonstrated that IL-13 is the primary upregulated cytokine in AD skin biopsy samples. A placebo-controlled phase 2b clinical trial evaluating the efficacy and safety of lebrikizumab, an IL-13 inhibitor, in AD demonstrated that, at 16 weeks, Eczema Area and Severity Index (EASI) 75 and Investigator’s Global Assessment (IGA) 0/1 were achieved by 60.6% and 44.6% of patients taking lebrikizumab at its highest dose (vs 24.3% and 15.3% of patients taking placebo, respectively). Moreover, treatment with lebrikizumab was associated with rapid improvement of pruritus and low rates of conjunctivitis (1.4%-3.8%). Another IL-13 monoclonal antibody, tralokinumab, was evaluated for safety and efficacy in moderate-to-severe AD. By week 12, among adults receiving 300 mg tralokinumab, 42.5% achieved EASI-75 and 26.7% achieved IGA 0/1 score (vs 15.5% and 11.8% in the placebo group, respectively). Both lebrikizumab and tralokinumab demonstrated acceptable safety profiles in AD (and non-AD) trials with adverse events often being comparable between treatment and control groups. Thus, IL-13 inhibitors may provide a safe and effective treatment alternative for patients with moderate-to-severe AD.

scholarly journals P149 The intracellular chloride channel 4 (CLIC4) plays an important role in systemic sclerosis fibroblast activation

Rheumatology ◽  
2021 ◽  
Vol 60 (Supplement_1) ◽  
Author(s):  
Christopher Wasson ◽  
Rebecca Ross ◽  
Ruth Morton ◽  
Jamel Mankouri ◽  
Francesco Del Galdo
Keyword(s):  
Systemic Sclerosis ◽  
Ion Channel ◽  
Smooth Muscle Actin ◽  
Chloride Ion ◽  
Dermal Fibroblasts ◽  
Cancer Associated Fibroblasts ◽  
Alpha Smooth Muscle Actin ◽  
Direct Role ◽  
Fibroblast Activation ◽  
Intracellular Chloride

Abstract Background/Aims  The intracellular chloride ion channel CLIC4 mediates the activation of cancer associated fibroblasts. Interestingly, systemic sclerosis (SSc) fibroblasts display a number of similar properties to cancer associated fibroblasts. Tissue fibrosis in SSc is driven by active fibroblasts (myofibroblasts). Therefore in this study we investigated the role of CLIC4 in SSc fibroblast activation. Methods  Dermal fibroblasts were obtained from full thickness skin biopsies from SSc patients (early-diffuse). RNA and protein were collected from the fibroblasts and CLIC4 transcript and protein levels were assessed by qPCR and western blot. SSc patient fibroblasts were treated with the chloride ion channel inhibitors NPPB and IAA-94. Results  CLIC4 was found to be expressed at significantly higher levels in SSc patients fibroblasts compared to healthy controls, at both the transcript (3.7 fold) and protein (1.7 fold) levels. Inhibition of the TGF-β signalling pathway led to reduced CLIC4 expression in SSc fibroblasts, confirming this pathway as the main driver of CLIC4 expression. Finally, treatment of SSc fibroblasts with small molecule inhibitors that target the channel led to reduced expression of the myofibroblast markers collagen type 1 and alpha-smooth muscle actin, suggesting a direct role for CLIC4 in SSc associated skin fibrosis. Conclusion  We have identified a novel role for CLIC4 in SSc myofibroblast activation, which further strengthen the similarities between SSc fibroblasts and cancer associated fibroblasts. Furthermore this study highlights this channel as a novel target for therapeutic intervention. Disclosure  C. Wasson: None. R. Ross: None. R. Morton: None. J. Mankouri: None. F. Del Galdo: None.

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