Predictive Drug Discovery Services
Respiratory Assays


REPROCELL Biopta is able to offer an extensive range of assays in fresh human lung tissue from healthy and diseased non-transplantable lungs or surgical residual tissues.

Such living human tissues allow REPROCELL to investigate the effects of drugs on the human respiratory systems in a number of ways including:

  • Bronchodilatation/bronchoconstriction
  • Ion channel function (and hence epithelial secretion)
  • Inflammatory responses
  • Epithelial drug absorption

Effects on respiratory function are already required to be assessed as part of the ICH S7A core battery of tests prior to human exposure. REPROCELL Biopta’s respiratory assays allow the safety of your compound to be determined far in advance of the clinical development phase. Additionally, we are able to procure diseased tissues, making it possible to carry out detailed safety studies on different patient populations, making comparisons that would otherwise be impossible. REPROCELL Biopta’s assays provide the optimum model for research into a range of conditions, such as COPD and asthma.

Whole lungs and resections of lung obtained from surgery are available fresh and can be used in a range of experimental setups such as tissue baths (shown below), wire myographs, Ussing chambers and ex vivo cultures including precision-cut lung slices.

 

Respiratory_assays_workflow

 

REPROCELL, under its Biopta brand pharmacology services, can investigate isolated fresh tissues from many regions of the lung, including upper airways, secondary or tertiary bronchi or parenchyma.

 

Airway Contractility

Using our organ bath or wire myographs for larger and smaller airways respectively, we are able to determine the bronchoconstrictive or bronchorelaxatory effects of a test compound in ex vivo human tissue. Figure 1 shows the effects of drugs on isolated rings of human bronchi; such studies can be conducted in large conducting airways or small bronchi, providing information on responses throughout the bronchial tree.

 

test-article_theophylline

 

Only human test systems truly reflect the human responses to drugs, even the most commonly used animal models of respiratory function, the guinea pig, fails to replicate all human responses (see Table 1). Human airways respond similarly to histamine; however, the responses to acetylcholine are markedly different. Using human tissues avoids uncertainties in decision-making based on animal models.

 

Species pD2 acetylcholine pD2 histamine
Human 4.56 5.31 ± 0.27
Guinea pig 5.89 5.31 ± 0.72

Precision Cut Lung Slices

Human and animal lung slices: a phenotypically-accurate functional assay system for screening and target validation

Precision-cut lung slices (PCLS) are increasingly in demand for the testing of xenobiotics, chemicals and cosmetics. By retaining both the structural and functional integrity of human or animal lungs, the method combines throughput with relevance, offering a phenotypically-accurate model of lung behaviour.

Moreover, Biopta’s unique access to healthy and diseased lung allows it to investigate drug effects in patients with COPD or asthma. Cross-species comparisons also allow the translation of preclinical animal data to the human situation to be assessed.

 

Experience-with-Precision-Cut-Lung-Slices-

 

Precision-cut lung slices:

  • Slices can be kept in culture for up to 7 days (Image 1)
  • Dozens of slices can be created from each donor or animal lung
  • The effect of test compounds on biomarker release, structural integrity, tissue viability or inflammatory processes can be evaluated (Image 2)
  • Biopta will work with you to design a protocol that meets your requirements.

Image 1:

PCLS_MTT-image

Image 2:

PCLS_LPS-image

 Download our Precision Cut Lung Slice Flyer (PDF)

To discuss a precision cut lung slice study with REPROCELL Biopta please contact us.


Respiratory channel function (ADME)

Lung or tracheal epithelial tissue can be assessed in the Ussing Chamber to determine changes in short circuit current (Isc) after exposure to your test compound. This information is particularly valuable when assessing treatments for conditions like cystic fibrosis where fluid movement across airway epethelium is disrupted resulting in thick mucus production.

Ussing-Chamber-Set-Up

REPROCELL Biopta has conducted a range of studies investigating specific ion channels in human or animal tracheal mucosa. Intact mucosa is dissected free from the trachea or primary bronchui and set up in Ussing chambers for measurement of functional responses. Figure 1 below shows the responses of human tracheal epithelium to the presence of increasing concentrations of the sodium channel blocker, amiloride.

Lung-ussing-image

 

Go to the Drug Discovery Assay Catalog »

Lung Parenchyma Culture

Through its extensive tissue network REPROCELL Biopta can obtain both healthy and diseased human lung samples for use in our range of ex vivo respiratory assays.

The parenchymal explant culture model provides an excellent platform to investigate and understand the mechanisms of lung diseases such as asthma and COPD. The model is also ideally suited for investigating the local effects of test compounds by being suitable for investigating a wide range of translational end points. The fresh explants contain the full complement of lung parenchymal cells in their native ratios and spacial orientation. This allows normal cell-cell communications and interactions to take place, replicating the local in vivo conditions of the lung.

An overview of the basic methodology is provided below in Figure 1.

Explant-Parenchymal-Culture-Methodology_V3

Upon receipt, the tissue is dissected free from the pleural membrane. Visible airways and blood vessels are also removed to produce the parenchyma explants (3-5mm3). The parenchyma explants are then maintained in culture, under physiological conditions, where the test conditions can be added. The orientation of the explants in culture provides the opportunity to study a wide range of experimental endpoints including (but not limited to) the following; the release of biomarkers/ mediators (culture media), the effects of test compounds on explant viability (culture media/explant), histopathological changes (explant), gene expression changes (explant) and changes in cell signalling mechanisms (explant).

Shown below in figure 2 is mean data obtained from experiments using 4 COPD donor lung samples. The ability of a range of standard of care treatments to modulate TNFalpha release from LPS stimulated explants was investigated. The standard of care drugs tested were the PDE 4 inhibitor Roflumilast, the glucocorticoid Fluticasone and the beta 2-adrenoceptor agonist Formoterol. Each treatment was tested alone and in combination with the others.

 

Parenchyma-culture-data-V2

 

Key benefits of REPROCELL Biopta’s human parenchymal explant model:

  • Native human tissue architecture is maintained in an in vitro assay format
  • Diseased vs non-diseased donor responses can be compared
  • Investigate mechanisms underpinning different donor responses
  • De-risk development and add commercial value by generating human proof of concept data prior to clinic
  • Measure multiple translational end-points from one experiment
  • Experiments can be customised according to each client’s need

Respiratory Assays in our Drug Discovery Assay Catalog

[B052] Bronchodilatation in secondary healthy human airways (Adrenoceptor – Isoprenaline)

[B053] Bronchoconstriction in secondary asthmatic human airways (Acetylcholine receptor)

[B054] Bronchodilatation in secondary asthmatic human airways (Adrenoceptor)

[B055] Bronchoconstriction in tertiary or lower healthy human airways (Prostanoid receptor)

[B056] Bronchodilatation in tertiary or lower healthy human airways (Adrenoceptor – Salbutamol)

[B057] Bronchoconstriction in tertiary or lower healthy human airways (Acetylcholine receptor)

[B058] Bronchodilatation in tertiary or lower healthy human airways (Adrenoceptor – Isoprenaline)

[B059] Bronchodilatation in secondary COPD human airways (Adrenoceptor)

[B060] Bronchoconstriction in denuded secondary healthy human airways (Acetylcholine receptor)

[B061] Bronchoconstriction in denuded secondary healthy human airways (Histamine receptor)

[B062] Ion channel function in healthy human tracheal mucosa (Adrenoceptors)

[B063] Ion channel function in healthy human tracheal mucosa (Sodium channels)

[B065] Bronchodilatation in tertiary or lower human airways (Adrenoceptors – Formoterol)

[B066] Bronchodilatation in tertiary or lower human airways (Adrenoceptors – Salbutamol)

[B067] Bronchodilatation in tertiary or lower asthmatic human airways (Adrenoceptor – Formoterol)

[B068] Bronchodilatation in tertiary or lower asthmatic human airways (Adrenoceptor – Salbutamol)

[B069] Bronchodilatation in tertiary or lower asthmatic human airways (Adrenoceptor – Salmeterol)

[B098] Bronchodilatation in healthy human secondary airways (VIP & PACAP receptors)

[B099] Bronchoconstriction in healthy human tertiary or lower airways (lopoxin, oxoeiconsanoid and resolvin E1)

Go to the Drug Discovery Assay Catalog »