Novel Therapies For Asthma Research Articles

Combination of androgen and estrogen improves asthma by mediating Runx3 expression.

Objective: Asthma is a chronic heterogeneous airway disease, and imbalanced T-helper type 1 (Th1) and Th2 cell-mediated inflammation contribute to its pathogenesis. Although it has been suggested that androgen and estrogen were involved in development of asthma, the underlying mechanisms remained largely unclear. Studies have demonstrated that Runx3 could promote naive CD4+ T cells to differentiate into Th1 cells. Hence, our study aimed to explore the potential regulatory mechanism of androgen and estrogen on asthma via modulating Runx3. Methods: First, clinical assessments and pulmonary function tests were conducted on 35 asthma patients and 24 healthy controls. The concentrations of androgen, estrogen, and androgen estrogen ratios were assessed in peripheral blood samples of asthma patients and healthy controls. Then, a murine asthma model was established to explore the effects of estrogen and androgen (alone or in combination) on asthma. Third, an in vitro assay was used to explore the mechanism of combination of androgen and estrogen in asthma. Results: We observed decreased androgen and increased estrogen levels in asthma patients compared with healthy controls. In mice with experimental asthma, there were increased serum concentrations of estrogen and decreased serum concentrations of androgen, intervention with combination of androgen and estrogen alleviated airway inflammations, increased Runx3 expressions and elevated Th1 differentiation. In CD4+ T cells co-cultured with bronchial epithelial cells (BECs), treatment with androgen plus estrogen combination promoted Th1 differentiation, which was mitigated by Runx3 knockdown in BECs and enhanced by Runx3 overexpression. Conclusion: These findings suggest that androgen estrogen combination modulate the Th1/Th2 balance via regulating the expression of Runx3 in BECs, thereby providing experimental evidence supporting androgen and estrogen combination as a novel therapy for asthma.

Schistosoma japonicum peptide SJMHE1 suppresses airway inflammation of allergic asthma in mice.

Helminths and their products can shape immune responses by modulating immune cells, which are dysfunctional in inflammatory diseases such as asthma. We previously identified SJMHE1, a small molecule peptide from the HSP60 protein of Schistosoma japonicum. SJMHE1 can inhibit delayed‐type hypersensitivity and collagen‐induced arthritis in mice. In the present study, we evaluated this peptide's potential intervention effect and mechanism on ovalbumin‐induced asthma in mice. SJMHE1 treatment suppressed airway inflammation in allergic mice, decreased the infiltrating inflammatory cells in the lungs and bronchoalveolar lavage fluid, modulated the production of pro‐inflammatory and anti‐inflammatory cytokines in the splenocytes and lungs of allergic mice, reduced the percentage of Th2 cells and increased the proportion of Th1 and regulatory T cells (Tregs). At the same time, Foxp3 and T‐bet expression increased, and GATA3 and RORγt decreased in the lungs of allergic mice. We proved that SJMHE1 can interrupt the development of asthma by diminishing airway inflammation in mice. The down‐regulation of Th2 response and the up‐regulation of Th1 and Tregs response may contribute to the protection induced by SJMHE1 in allergic mice. SJMHE1 can serve as a novel therapy for asthma and other allergic or inflammatory diseases.

Type 1 alveolar epithelial cell-derived chemokine CCL19 promotes airway inflammation

Abstract Allergic asthma is largely driven by the actions of allergen-specific CD4+T helper cells. Following allergen inhalation, these cells are induced by the interaction of naïve CD4+ T cells with antigen-bearing dendritic cells (DCs) in lung-draining lymph nodes. The molecular mechanisms underlying the induction of T helper cells have been well studied, and include the migration of allergen-bearing DCs to lung-draining lymph nodes in a manner dependent on the chemokine receptor, CCR7. However, comparatively little is known regarding how these effector T cells become activated following their subsequent encounter with allergen in the lung. This knowledge would be useful in the design of novel therapies for asthma, especially for neutrophilic asthma, which is refractory to inhaled glucocorticoids. We recently noticed that lung DCs migrating to lymph nodes originate from the large airways, whereas DCs associated with small airways, parenchyma and subpleural areas are non-migratory. Video confocal analysis of precision cut lung slices revealed that these non-migratory DCs form tight interactions with OVA-specific T cells and stimulate T cell receptor signaling, as measured by activation of a Nur77-gfpreporter gene. Furthermore, Ccl19, which encodes a CCR7-ligand, is highly expressed by type 1 alveolar epithelial cells, and in mouse model of asthma, airway inflammation was significantly diminished in CCL19-deficient mice compared with WT mice. Together, our findings suggest that CCL19 produced by type I epithelial cells acts to position lung DCs in the distal area of the lung where they interact with effector T cells, thereby promoting allergic airway inflammation.

The Catalytic α Subunit of cAMP‐Dependent Protein Kinase Regulates Formoterol‐Induced Gene Expression in BEAS‐2B Human Bronchial Epithelial Cells

RationaleThe β2‐adrenoceptor is a GPCR that classically couples to Gsα, the activation of adenylyl cyclases and the formation of cAMP. Through distinct signalling complexes often containing cAMP‐dependent protein kinase (PKA), cAMP mediates a myriad of functional responses including the regulation of gene expression. We have shown previously that in the BEAS‐2B human airway epithelial cell line, long acting β2‐adrenoceptor agonists (LABAs) induce cAMP response element (CRE) dependent transcription, which we suggest contributes to their therapeutic activity in obstructive lung diseases. In the current study we have extended those findings by identifying the PKA catalytic subunit responsible for mediating gene expression induced by the LABA, formoterol.MethodsMicroarray, RT‐PCR, western blotting and gene silencing were used to identify and implicate PKA catalytic subunits in the regulation of LABA‐induced gene expression. A CRE reporter construct stably transfected into BEAS‐2B cells was used for this purpose together with an assessment of CREB phosphorylation and an analysis of a panel of cAMP‐inducible genes (NR4A3, CRISPLD2, RGS2, NR4A2, CD200 and IL‐6).ResultsMicroarray‐based gene expression profiling indicated that BEAS‐2B cells express mRNA transcripts for PRKACA (Cα), PRKACB (Cβ) and PRKX (Cx) whereas PRKACG (Cg) was absent. These data were confirmed at the protein level by western blotting. siRNAs were optimized to achieve 80 to 90% knockdown of PRKACA, PRKACB and PRKX, which was reproduced at the protein level. Under these conditions, silencing PRKACA was associated with a significant (~50%) abrogation of formoterol‐induced CRE reporter activation, CREB phosphorylation and gene expression. No additional effect was seen when siRNAs that target PRKACB or PRKX were used in combination with those directed towards PRKACA.ConclusionsThese data show that Cα is an essential cAMP signalling component controlling formoterol‐induced gene transcription in BEAS‐2B cells. The incomplete inhibition produced under conditions of 80–90% Cα knockdown implies that this PKA catalytic subunit is present in significant excess of that required to maximally mediate cAMP‐induced gene expression. This is an initial first step at identifying cAMP signalling components (adenylyl cyclases, scaffold proteins, phosphodiesterases and effectors) that control LABA‐induced functional responses in airway structural cells. We propose that identifying these components and their assembly into multiprotein signalling complexes could provide new opportunities for the development of novel therapies for asthma and chronic obstructive pulmonary disease.Support or Funding InformationAlberta Lung Association (ALA)

Editorial introductions

Editorial introductions

INSIDE INDUSTRY

Sanford-Burnham and Daiichi Sankyo form alliance to study novel drug targets in cardiovascular metabolic diseases. Sigma-Aldrich® to launch Austrianova's Cell-in-a-Box® Kit for research use. ASLAN Pharmaceuticals signs agreement with CSL to develop novel therapy for asthma. Self-assembling peptides for bone regeneration and drug delivery. Verisante Technology, Inc. receives the 2014 North American Technology Innovation Leadership of the Year Award from Frost & Sullivan. Agilent Technologies partners Women in Science, Engineering and Technology in Korea. Mundipharma and Genewel sign agreement across key global markets for licensing and promotion of Medifoam®. Janssen collaborates with ViiV Healthcare to develop two-drug single tablet regimen for the maintenance treatment of people living with HIV. AbbVie opens expanded manufacturing facility in Sligo, Ireland. AMRI to Acquire Oso Biopharmaceuticals Manufacturing. Merck to collaborate with Sysmex Inostics on a blood-based RAS biomarker test. Igenbio establishes direct presence in Japan. Mesoblast accelerates plans for commercial manufacturing operations in Singapore. Celerion and Korean Drug Development Fund form collaboration to strengthen early phase clinical research in South Korea. Dr. Carolyn Mountford, University of Newcastle, Australia receives Agilent Technologies Thought Leader Award. PHARM Connect – The business summit covering a full industry. New topics for the Next Generation Sequencing & Single Cell Genomics Asia Congress confirmed. Registration opened for Oxford Global's 6th Annual Next Generation Sequencing Congress. Registration opened for Oxford Global's 4th Annual Next Generation Sequencing Asia Congress.

Effect of inhibiting the phosphorylation of signal transducer and activator of transcription-3 on Th2 cell-mediated airway inflammation and airway remodeling in a mouse model of asthma

Objective To study the effect of inhibiting the phosphorylation of signal transducer and activator of transcription-3 (STAT3) on Th2 cell-mediated airway inflammation and airway remodeling, and to explore the role of STAT3 in the pathophysiology of bronchial asthma. Methods Forty Balb/c mice were randomly divided into control group(n=10), asthma group(n=10), AG490 by intraperitoneal group (n=10), and AG490 by inhalation group (n=10). The mice were sensitized with ovalbumin to establish the asthmatic model.The histological changes were evaluated by means of HE staining, while total broalchial wall thickness(Wat)and smooth muscle thickness(Wam) were mea-sured by using image analysis system.The percentages of collagen deposition were detected by way of Masson's trichrome staining; the bronchoalveolar lavage fluid (BALF) were collected, the total cell and the cell differentials were counted, the levels of IL-4, IL-5 in BALF were measured by enzyme-linked immunosorbent assay; the lung tissue extracts were analyzed for phosphorylation of STAT3(p-STAT3)by Western blot.The SPSS 13.0 software was used to analyze the data. Results 1.The histological changes by HE staining showed that less inflammatory cells infiltration in airway and around the pulmonary vascular in AG490 administration groups compared with asthmatic group.Wat, Wam and the percentages of collagen deposition in AG490 administration groups was significantly lower than that in asthmatic group(F=49.5, 41.7, 58.2, all P< 0.05).2.The level of p-STAT3 in the lung of AG490 administration groups were significantly lower than those in asthmatic group(F=34.17, P< 0.05).3. The total cells and eosinophils amounts in BALF of AG490 administration groups were significantly lower than those in asthmatic group(F=42.5, 64.7, all P< 0.05). The levels of IL-4, IL-5 in BALF of AG490 administration groups were significantly lower than those in asthmatic group, respectively (F=39.2, 75.1, all P<0.05). Conclusions STAT3 signaling pathway is pivotal in Th2 cell-mediated airway inflammation and airway remodeling in asthmatic models, and AG490 can ameliorate airway inflammation and airway remodeling efficiently by inhibiting the phosphorylation of STAT3, and targeting this signaling pathway may be a novel therapy for asthma. Key words: Asthma; Th2 type cell; Airway inflammation; Airways remodeling; Signal transducer and activator of transcription-3

Editorial introductions

Editorial introductions

Synthetic Response of Stimulated Respiratory Epithelium: Modulation by Prednisolone and iKK2 Inhibition

BackgroundThe airway epithelium plays a central role in wound repair and host defense and is implicated in the immunopathogenesis of asthma. Whether there are intrinsic differences between the synthetic capacity of epithelial cells derived from subjects with asthma and healthy control subjects and how this mediator release is modulated by antiinflammatory therapy remains uncertain. We sought to examine the synthetic function of epithelial cells from different locations in the airway tree from subjects with and without asthma and to determine the effects of antiinflammatory therapies upon this synthetic capacity. MethodsPrimary epithelial cells were derived from 17 subjects with asthma and 16 control subjects. The release of 13 cytokines and chemokines from nasal, bronchial basal, and air-liquid interface differentiated epithelial cells before and after stimulation with IL-1β, IL-1β and interferon-γ, or Poly-IC (Toll-like receptor 3 agonist) was measured using MesoScale discovery or enzyme-linked immunosorbent assay, and the effects of prednisolone and an inhibitor of nuclear factor κ-B2 (IKK2i) were determined. ResultsThe pattern of release of cytokines and chemokines was significantly different between nasal, bronchial basal, and differentiated epithelial cells but not between health and disease. Stimulation of the epithelial cells caused marked upregulation of most mediators, which were broadly corticosteroid unresponsive but attenuated by IKK2i. ConclusionSynthetic capacity of primary airway epithelial cells varied between location and degree of differentiation but was not disease specific. Activation of epithelial cells by proinflammatory cytokines and toll-like receptor 3 agonism is attenuated by IKK2i, but not corticosteroids, suggesting that IKK2i may represent an important novel therapy for asthma.

Eosinophil apoptosis and clearance in asthma.

Asthma is an increasingly common respiratory condition characterized by reversible airway obstruction, bronchial hyper-responsiveness and airway inflammation with a clear unmet need for more effective therapy. Eosinophilic asthma is a phenotype of the condition that features increased blood or sputum eosinophils whose numbers correlate with disease severity. Several lines of evidence are now emerging, which implicate increased persistence of eosinophils in the lungs of patients with asthma as a consequence of inhibition of and defects in the apoptotic process, together with impaired apoptotic cell removal mechanisms. This article will update our knowledge of the mechanisms controlling eosinophil apoptosis and clearance, together with evidence implicating defects in apoptosis and pro-inflammatory cell removal in asthma. Recent developments in novel therapies for asthma that target eosinophil apoptotic and/or clearance pathways will also be discussed.

Editorial introductions

Editorial introductions

Emerging therapies for severe asthma

Many patients with asthma have poorly controlled symptoms, and particularly for those with severe disease, there is a clear need for improved treatments. Two recent therapies licensed for use in asthma are omalizumab, a humanized monoclonal antibody that binds circulating IgE antibody, and bronchial thermoplasty, which involves the delivery of radio frequency energy to the airways to reduce airway smooth muscle mass. In addition, there are new therapies under development for asthma that have good potential to reach the clinic in the next five years. These include biological agents targeting pro-inflammatory cytokines such as interleukin-5 and interleukin-13, inhaled ultra long-acting β2-agonists and once daily inhaled corticosteroids. In addition, drugs that block components of the arachidonic acid pathway that targets neutrophilic asthma and CRTH2 receptor antagonists that inhibit the proinflammatory actions of prostaglandin D2 may become available. We review the recent progress made in developing viable therapies for severe asthma and briefly discuss the idea that development of novel therapies for asthma is likely to increasingly involve the assessment of genotypic and/or phenotypic factors.

Induction of regulatory T cells by a novel immunoregulatory therapy suppresses the development of allergic airways disease (97.10)

Abstract Evidence suggests that exposure to Streptococcus pneumoniae may be protective against asthma. We have identified key S. pneumoniae components that form an immunoregulatory therapy (IRT) and suppress the development of allergic airways disease (AAD) using mouse models. AAD was induced in adult BALB/c mice by intraperitoneal sensitization and intranasal challenge with ovalbumin. At the time of sensitization, the IRT was administered intratracheally. IRT treatment suppressed hallmark features of AAD including eosinophil influx, local and systemic Th2 cytokines, airways hyperresponsiveness and mucus hypersecretion. A time course analysis showed that the IRT induced CD4+CD25+Foxp3+ Tregs in the lymph nodes 4 days post treatment. This was associated with increased IL-2, TGF-β and Foxp3 expression in the lungs. IRT treatment also increased the proportion of CD4+CD25+/- cells expressing Foxp3 in the lungs and expression of markers associated with Treg function, during the induction of AAD. Anti-CD25 administration 3 days before IRT treatment abolished the suppressive effects on AAD. However, anti-CD25 administration 9 days after IRT treatment had no effect. This indicated a requirement for IL-2 signalling via CD25 (IL-2R) during the induction of Tregs by the IRT treatment. In conclusion, a S. pneumoniae-based IRT induces CD25-dependent Tregs, suppresses the development of AAD and may be useful as a novel therapy for asthma.

Streptococcus pneumoniae vaccine, Prevenar, utilises Tregs to suppress Asthma (140.2)

Abstract Our recent finding that exposure to Streptococcus pneumoniae may suppress pro-asthmatic responses led us to investigate whether the current human S. pneumoniae vaccines may be used to suppress asthma using mouse models of ovalbumin-(OVA)-induced allergic airways disease (AAD). AAD was induced by intraperitoneal sensitisation and intranasal challenge with OVA. At the time of OVA sensitisation, Prevenar or Pneumovax were delivered intranasally either with or without CpG. Prevenar, but not Pneumovax, suppressed hallmark features of AAD, including eosinophil influx, OVA-specific T helper (Th) 2 cytokine release, mucus hypersecretion, total serum IgE and airways hyperresponsiveness. We then investigated whether regulatory T cells (Tregs) were important in the suppression of AAD. Prevenar increased the number of CD4+CD25+FoxP3+ expressing Tregs in the lungs, lymph nodes and spleens. The release of immuno-suppressive cytokines IL-10 and TGF-â from lymph nodes was reduced. However, Prevenar increased expression of CD103 and CTLA-4 on CD4+CD25+FoxP3+ cells. Proliferation assays demonstrated that Tregs suppressed allergic effector T cell proliferation regardless of Prevenar treatment. In conclusion, a currently available human vaccine increases the infiltration of Tregs and suppresses AAD, and may be useful as a novel therapy for asthma.

Discovery of Novel Therapies for Asthma Requires Suitable and Relevant Disease Models

Discovery of Novel Therapies for Asthma Requires Suitable and Relevant Disease Models

Investigation of the Molecular Mechanisms Preceding PDE4 Inhibitor-Induced Vasculopathy in Rats: Tissue Inhibitor of Metalloproteinase 1, a Potential Predictive Biomarker

Phosphodiesterase (PDE) 4 inhibitors are a class of drugs that can provide novel therapies for asthma and chronic obstructive pulmonary disease. Their development is frequently hampered by the induction of vascular toxicity in rat mesenteric tissue during preclinical studies. Whereas these vascular lesions in rats have been well characterized histologically, little is known about their pathogenesis and in turn, sensitive and specific biomarkers for preclinical and clinical monitoring do not exist. In order to investigate the early molecular mechanisms underlying vascular injury, time-course studies were performed by treating rats for 2-24 h with high doses of the PDE4 inhibitor CI-1044. Transcriptomics analyses in mesenteric tissue were performed using oligonucleotide microarray and real-time RT-PCR technologies and compared to histopathological observations. In addition, protein measurements were performed in serum samples to identify soluble biomarkers of vascular injury. Our results indicate that molecular alterations preceded the histological observations of inflammatory and necrotic lesions in mesenteric arteries. Some gene expression changes suggest that the development of the lesions could follow a primary modulation of the vascular tone in response to the pharmacological effect of the compound. Activation of genes coding for pro- and antioxidant enzymes, cytokines, adhesion molecules, and tissue inhibitor of metalloproteinase 1 (TIMP-1) indicates that biomechanical stimuli may contribute to vascular oxidant stress, inflammation, and tissue remodeling. TIMP-1 appeared to be an early and sensitive predictive biomarker of the inflammatory and the tissue remodeling components of PDE4 inhibitor-induced vascular injury.

Adenosine Receptors As Promising Therapeutic Targets for Drug Development in Chronic Airway Inflammation

A growing body of evidence has emerged in support of a pro-inflammatory role for adenosine in the pathogenic mechanisms of chronic inflammatory disorders of the airways such as asthma and COPD. The demonstration that adenosine enhances mast cell allergen-dependent activation, the notion that elevated levels of adenosine are present in chronically inflamed airways, and the results from exposure studies of nebulised adenosine showing dose-dependent bronchoconstriction in subjects with asthma and COPD, emphasise the importance of adenosine in the initiation, persistence and progression in these common inflammatory disorders of the airways. Adenosine exerts its manifold biological activities by interacting with at least four adenosine receptor subtypes. Selective activation or blockade of these sites is being exploited by the pharmaceutical industry in an attempt to generate novel therapies for asthma and COPD. This review article intends to address the potential role of adenosine in asthma and to put forward the idea that drugs that have been developed to selectively activate or downregulate adenosine receptor subtypes may constitute a considerable advance in the management of chronic airway inflammation.

Adenosine signalling in airways

Current research endeavours indicate that adenosine elicits strong inflammatory responses in the lung and might be involved in the pathogenic mechanisms of chronic inflammatory disorders of the airways such as asthma and chronic obstructive pulmonary disease (COPD). The contribution of adenosine-associated effector mechanisms to the initiation, persistence and progression of the inflammatory response is highly heterogeneous and is dictated by the expression pattern of four different adenosine receptors. Selective activation or blockade of these sites can therefore be exploited in an attempt to generate novel therapies for asthma and COPD. In addition, an important development is the use of adenosine (or AMP) as a diagnostic test for discriminating asthma from COPD, and as an accurate biomarker to monitor corticosteroid requirements in asthma. It is likely that therapies interfering with adenosine signalling in the airways will offer a considerable advance in the management of asthma and COPD.

Understanding the pathophysiology of severe asthma to generate new therapeutic opportunities

Although asthma is defined in terms of reversibility of airflow obstruction, as the disease becomes more severe and chronic, it adopts different characteristics, including a degree of fixed airflow obstruction and corticosteroid refractoriness. Underlying these phenotypes is evidence of airway wall remodeling, which should be distinguished from the increase in smooth muscle linked to airways hyperresponsiveness. Aberrant epithelial-mesenchymal communication leads to a chronic wound scenario, which is characterized by activation of the epithelial-mesenchymal trophic unit, epithelial damage, the laying down of new matrix, and greater involvement of neutrophils in the inflammatory response. In allergic asthmatic patients who remain symptomatic despite high-dose corticosteroid therapy, blockade of IgE with omalizumab confers appreciable clinical benefit. Chronic severe asthma is also accompanied by a marked increase in TNF-alpha production that might contribute to corticosteroid refractoriness. Based on this, TNF blockade with the soluble fusion protein entanercept produces improvement in asthma symptoms, lung function, and quality of life paralleled by a marked reduction in airways hyperresponsiveness. Identification of novel susceptibility genes, such as a disintegrin and metalloprotease 33 (ADAM33), will provide further targets against which to direct novel therapies for asthma, especially at the more severe end of the disease spectrum.

Adenosine receptors: promising targets for the development of novel therapeutics and diagnostics for asthma

Interest in the role of adenosine in asthma has escalated considerably since the early observation of its powerful bronchoconstrictor effects in asthmatic but not normal airways. A growing body of evidence has emerged in support of a proinflammatory and immunomodulatory role for the purine nucleoside adenosine in the pathogenic mechanisms of chronic inflammatory disorders of the airways such as asthma. The fact that adenosine enhances mast cell allergen-dependent activation, that elevated levels of adenosine are present in chronically inflamed airways, and that adenosine given by inhalation cause dose-dependent bronchoconstriction in subjects with asthma emphasizes the importance of adenosine in the initiation, persistence and progression of these common inflammatory disorders of the airways. These distinctive features of adenosine have been recently exploited in the clinical and research setting to identify innovative diagnostic applications for asthma. In addition, because adenosine exerts its multiple biological activities by interacting with four adenosine receptor subtypes, selective activation or blockade of these receptors may lead to the development of novel therapies for asthma.