Respiratory Research Group - Honours projects available in 2010

An Honours project undertaken in this lab would be administered by the Discipline of Pharmacology.

1. How different are fibroblasts from various lung diseases?

Supervisor + contact details:

• Dr Brian Oliver

Fibroblasts are key cells in lung biology, and are vital for maintaining normal lung homeostasis. In this project you would investigate if lung fibroblasts from people with different respiratory diseases proliferate at different rates, and determine if the inflammatory profile of these cells are different. Once you have established the baseline characteristics of these cells you would then investigate if currently used respiratory drugs are effective at inhibiting these processes. This project will use a range of cutting edge cellular and molecular biological techniques (such as western blotting and real time PCR) located within our laboratories in the Discipline of Pharmacology.

2. Can the common cold virus kill you?

Supervisor + contact details:

• Dr Brian Oliver

We all suffer from the common cold, but the viruses that cause the symptoms of the common cold can be potentially life threatening to certain people. The reason why the common cold virus, rhinovirus, is potentially life threatening is that it is responsible for the majority of exacerbations of chronic obstructive pulmonary disease (COPD). The reason why rhinovirus-induced exacerbations occur is not known, however it is thought that rhinovirus-induced cytokine release contributes to this process. This project will assess if/and why lung cells from people with COPD have altered responses to rhinovirus (e.g. cytokine release). It will also assess if medications used to treat COPD can effect rhinovirus-induced changes.

3. Can viruses stop b₂-agonists from working?

Supervisor + contact details:

• Dr Brian Oliver

Asthma exacerbations cause substantial morbidity, mortality and health care costs. Asthma exacerbations are characterised by worsening asthma symptoms and a fall in lung function, and contribute substantially to the cost and burden of asthma. In NSW alone there were 22,942 ED visits for asthma of which 42% were admitted to hospital in 2007. The annual cost per person for hospital admissions can reach $23,766. In addition to the direct medical costs, the costs of absenteeism are thought to account for 50% of the total cost of treating asthma in Australia.

Rhinovirus infection cause asthma exacerbations. Viruses are identified in 80% of asthma exacerbations in children and 45-80% in adults. Rhinovirus, which causes the majority of common colds, is responsible for at least half of asthma exacerbations.

b₂-agonist efficacy is reduced during viral exacerbations. Under normal circumstances, asthmatic airway obstruction improves characteristically briskly in response to inhaled b₂-adrenergic agonists (e.g., salbutamol), which are the most commonly used asthma therapy in most Western countries. Loss of response to b₂-agonist is often the trigger for hospital presentation, and may be life-threatening. During an exacerbation there is objective evidence that an impaired response to b₂-agonists occurs. We believe that the mechanisms by which viral exacerbations occur are distinct from those operating when asthma control is inadequate, since exacerbations can occur in otherwise well controlled asthmatics.

Surprisingly, given the importance of rhinovirus infections in acute exacerbations of asthma the mechanism by which rhinovirus induces an impaired response to b₂-agonists is not known.

Hypothesis

The impaired response to short-acting b2-agonists which occurs with viral infections in asthmatic patients is due to b2-receptor dysfunction.

preliminary data

We have data which shows that factor(s) released from RV infected bronchial epithelial cells cause decreased β₂-agonist-induced cAMP in airway smooth muscle. However there are several unanswered questions:

1. We also don't know if the impaired β₂-agonist-induced cAMP can be restored by the addition of more β₂-agonist
2. We don't know how quickly the factor is produced in rhinovirus infected epithelium
3. We don't know if this phenomena is restricted to rhinovirus or is common to other viruses.
4. We don't know if other causes of asthma exacerbations eg allergens also result in impaired β₂-agonist-induced cAMP.

The aims of this project would be to choose 1 or 2 of the research ideas from the above list and investigate this for your honours. You would also evaluate weather current asthma therapeutics effect experimental outcomes. You would use an in vitro tissue culture model to investigate your aims. Typical techniques would include primary cell culture, real time PCR, FACS and western blotting.

4. Does Tumstain regulate inflammation asthma?

Supervisor + contact details:

• Dr Brian Oliver

Asthma is a complex and mulit-factorial disease of the airways in which both inflammation and remodelling are important. Remodelling includes alterations in the extracellular matrix (ECM). Inflammation is largely treatable through the use of current therapeutics such as corticosteroids, however upon cessation of treatment inflammation reoccurs. We have discovered that an endogenous anti-inflammatory and antiangiogenic molecule, tumstatin, derived from the ECM, is absent from the airways of asthmatics.

Tumstatin is the non-collagenous (NC)-1 domain of the collagen IV a3 chain. Collagen IV, a major constituent of the basement membrane, exists in the ECM when any three of the six collagen IV a chains combine. We have identified the presence of the NC-1 domains of all six collagen IV a chains in non-asthmatic human lung sections, however tumstatin was absent in all asthmatic lung sections irrespective of treatment, age and severity.

ECM proteins are important in the regulation of the inflammatory process, with fragments of collagen having both pro- and anti-inflammatory properties. Whilst tumstatin is known as an angiogenic inhibitor, it also has anti-inflammatory properties which may be mediated via modulation of cytokines and/or chemokines produced by inflammatory cells.

Hypothesis

That the absence of tumstatin in the asthmatic airway is a major contributory factor to the underlying pathophysiological mechanism through which asthma occurs.

Specific aims to determine if the lack of tumstatin leads to an enhanced inflammatory response in the asthmatic airway by a) directly altering the basal or stimulated release of inflammatory chemokines and cytokines (IL6, IL8, eotaxin, VEGF and Ang-1) from inflammatory cells.

Methods

In this project lymphocytes will be isolated from peripheral blood and human lung tissue. You will then use an in-vitro cell culture system to evaluate how these cells respond to tumstatin. This project will use the latest molecular biological techniques (eg siRNA, real time PCR, ELSIA western bloting).

Outcomes and Significance: Airway inflammation, a cardinal characteristic of asthma, is usually well controlled by corticosteroids. However, this approach only controls the symptoms and not the underlying cause of the disease. Our novel observation that tumstatin, an antiinflammatory and antiangiogenic molecule is absent from asthmatic airways presents an opportunity for innovative ways of targeting inflammation in asthma.

5. Pulmonary lymphangioleiomyomatosis (LAM)

Supervisor + contact details:

• Dr Lyn Moir

Pulmonary lymphangioleiomyomatosis (LAM) is a progressive and usually fatal rare lung disease that affects almost exclusively young females and for which there is no treatment. The average age of onset for LAM disease is between 25 and 40 years of age. During the progression of the disease abnormal smooth muscle-like cells known as LAM cells invade the healthy lung tissue and obstruct the airways, blood and lymph vessels, preventing the lungs from functioning properly. Scientific research over the last 10 years has shown that LAM is associated with genetic mutations in the tuberous sclerosis genes, tuberous sclerosis complex 1 (TSC1) and TSC2. In particular, loss of function of TSC2 results in phenotypic manifestation of the disease. Dysfunction of TSC2 has been associated with altered cellular function including cell growth, migration and invasiveness. However, the cellular and molecular mechanisms of LAM disease are not well understood.

The aim of this study is to investigate the role of TSC2 in the enhanced cellular functions associated with LAM. By understanding the role of TSC2 we will enhance our knowledge of the basic mechanisms of LAM disease.

To investigate the aim you would use an in vitro tissue culture model. Other typical techniques include gel electrophoresis (Western blotting), flow cytometry, immunohistochemistry and real-time PCR.