Respiratory

Respiratory

Respiratory diseases have been identified to be an immense health burden worldwide with approximately 1 billion persons suffering from chronic respiratory conditions.
Estimates suggest that:
Asthma and chronic obstructive pulmonary disease (COPD) are two long-term inflammatory conditions of the lung airways, both related with exposure to environmental factors (like pollution and allergens) for asthma and smoking for COPD [1, 2,].
Asthma is a chronic airway inflammatory disease characterised by airway hyperresponsiveness leading to breathlessness associated with wheezing and cough [3]. It is frequently known to coexist with atopic diseases, particularly allergic rhinitis [4].
The GOLD initiative defines COPD as “a disease state characterized by airflow limitation that is not fully reversible. The airflow limitation is usually both progressive and associated with an abnormal inflammatory response of the lungs to noxious particles or gases.” [5]. The American Thoracic Society defines COPD in terms of chronic bronchitis and emphysema.13 Chronic bronchitis is characterized by the clinical symptoms of excessive cough and sputum production; emphysema refers to chronic dyspnea, resulting from enlarged air spaces and destruction of lung tissue [6].
The pharmaceutical approach for asthma and COPD is still relatively ineffective and based on the administration of drugs that will control the symptoms. These conditions may worsen over the years, requiring periodic hospitalization (asthma) or evolving by developing a cardiovascular-associated disease or lung cancer (COPD).
Idiopathic pulmonary fibrosis (IPF) is the commonest interstitial lung disease (ILD) and is characterised by progressive scarring of the lungs. Impaired pulmonary would healing response following lung injury coupled with excessive production of collagens leads to a pathogenic lung fibrosis [7].
  1. Papi A et al. The Lancet. 2018, Feb 24.
  2. King TP. Clin Transl Med. 2015; 4: 26.
  3. Quirt J et al. Allergy Asthma Clin Immunol. 2018; 14(Suppl 2): 50.
  4. Bourdin A et al. Thorax. 2009;64(11):999–1004
  5. Pauwels RA et al. Am J Respir Crit Care Med. 2001; 163: 1256–1276
  6. Standards for the diagnosis and care of patients with chronic obstructive pulmonary disease.  Am J Respir Crit Care Med. 1995; 152 (5 pt 2): S77–S121
  7. Wilson MS and Wynn TA. Mucosal Immunol. 2009 Mar; 2(2): 103–121.
Cellomatics Biosciences Ltd. provides expertise in providing well validated preclinical airway models to investigate the effects of compounds in a variety of pathological settings including inflammation, gel contraction, oxidative stress, mast cell degranulation, extracellular matrix deposition (ECM) and epithelial to mesenchymal transition. These models are suitable for low to medium throughput studies.

1. Asthma and Airway Inflammation

2. Allergic Rhinitis

3. COPD

4. Mast Cell Degranulation

HMC1.2 cells were treated with Calcium Ionophore and Compound 48/80 in the presence or absence of Protamine and Dexamethasone. Vehicle control included 0.1% DMSO in culture media. The beta-hexosaminidase alpha (HEXA) was analysed in the cell lysates by ELISA. An increase in HEXA was observed with CaI and 48/80 when compared to vehicle control/ untreated. These levels of HEXA reduced when treated with inflammatory inhibitors such as Protamine and Dexamethasone.

5. Pulmonary Fibrosis

In vitro models – Pulmonary Fibrosis

A. Measurement of inflammatory mediators
  • Cytokines relevant to mechanisms of pulmonary fibrosis : TGF-beta, Connective tissue growth factor (CTGF, CCN2), PDGF, IGF, IL4, IL13, IFN- gamma, TNF-alpha, IL17, Oncostatin M, IL10
  • Chemokines: MCP1, CCL18, MIP1-alpha, CXCL12
B. Measurement of oxidative stress
  • 8-oxo-dG
  • Superoxide dismutase assay
  • Intracellular ROS levels
  • Hydrogen peroxide
C. Collagen Gel contraction assay
  • To assess contractile properties of target cells i.e. smooth muscle cells/fibroblasts
D. Increased ECM deposition
  • Measure collagen synthesis (indicator of fibrosis)
    1. Human lung fibroblasts treated with TGF-beta in the presence or absence of test compounds
    2. Measure collagen I and IV levels in cell culture supernatants
  • Measure hydroxyproline content (major component of collagen)
    1. Human lung fibroblasts treated with TGF-beta in the presence or absence of test compounds
    2. Measure hydroxyproline levels in cell culture supernatants
  • Measure other ECM proteins
    1. MMP1
    2. PAI-1
E. Epithelial Mesenchymal Transition (EMT)
  • Immunostaining for fibronectin in human bronchial epithelial cells following trans-differentiation with TGF-beta in the presence or absence of test compounds

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Cellomatics Biosciences Ltd. is a specialised Contract Research Organisation (CRO) providing bespoke preclinical laboratory based services.

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