Research

All proceeds from '65km for Cystic Fibrosis' will be directed to  CF research or research that improves the quality of CF care in hospitals as this is known to be essential for good health outcomes.  

To date we have funding the following projects:

2016 - Monash Foundation

Is implementation of the 2006 Australasian Clinical Practice Guidelines for nutrition in cystic fibrosis associated with improvements in nutritional status?  

Associate Professor David Armstrong - Monash Childrens

$ 22,200 grant

We are interested in whether the Australasian Clinical Practice Guidelines for nutrition in cystic fibrosis, which were implemented in 2006, are associated with improvements in the nutritional status of children participating in the Australian Cystic Fibrosis Data Registry (ACFDR).  Nutrition in children with CF is very important because poor nutrition is a risk factor for early death.  However, poor nutrition remains a problem in CF because  of increased energy requirements and problems with fat and protein absorption.  To help address this, Australasian guidelines for dietitians for the nutritional care of CF children were introduced in 2006.  These make suggestions for dietitians to help children with CF eat well and grow optimally.  However, whether the nutritional status in children with CF has improved since the introduction of the guidelines has not been evaluated.  This is what we will do in this study.  We will extract the nutritional status data of children with CF in the ACFDR from 1998.  We will compare growth data before 2006 and after 2006 in these children using statistical modelling.  In addition, we will explore whether there were problems implementing the guidelines by conducting an online survey among stakeholder groups (such as dietitians, doctors, nurses and patents).  We expect the find improved nutritional status in children after the introduction of the guidelines.  We also expect to find out what the enablers and barriers to implementing the guidelines were.  This will help inform implementation strategies for the soon to be released updated Australasian nutrition guidelines for CF

Cystic Fibrosis (CF) questionnaire assessing disease knowledge in adolescents with CF

Dr. Moya Vandeleur - Monash Health

$ 30,271 grant

Transition to adult care is a major life event for young people with cystic fibrosis (CF).  Many studies have reported that in part a successful transition relies on imparting sufficient knowledge about CF; the disease itself and treatment plans.  A higher level of knowledge has been correlated with better disease self-management and treatment compliance.  At Monash, we have a roust transition process as paediatric patients transfer to co-located adult service or from Royal Children's Hospital (RCH).  However, relatively little is known about disease specific knowledge of our adolescent patients with CF and their parents.  As part of an initiative to improve the CF transition processes at Monash Children's Hospital (MCH) and RCH, we aim to develop a reliable questionnaire to investigate general knowledge about CF in both adolescents and their parents.  To validate its use in a larger population we plan to collaborate and pilot the questionnaire at both MCH and the RCH.  Potential participants include all young people with CF aged 11 - 18 years (and their parents / careers) attending the CF out patient service at MCH and RCH.  The adolescent and their parent / carer will complete the questionnaire following a clinic visit electronically using a handheld computer.  

The purpose of this project is to improve the CF transition process at MCH and RCH by developing a questionnaire that will evaluate knowledge in transitioning young people and measure the effectiveness of the CF team in communicating information about cystic fibrosis.

2015  MCRI

How does activation of PAR1 (Protease-activated receptor-1) modify the response of lung immune cells in cystic fibrosis?

Associate Professor Phil Sutton 

$       TBC   Grant

Progress Report - November 2016

Protease Activated Receptor 1 (PAR1) is a molecule found on the surface of many different cell types, including those of the immune system, which has been shown to be able to change the severity of inflammation in a range of different diseases.  We have been examining whether PAR1 might play a role in changing the severity of inflammation in the CF lung and, if so, whether it might therefore provide a novel therapeutic target.  The studies we have performed so far suggest that activation of PAR1 might modify the inflammatory response to pathogenic bacterial infection in lung immune cells some children with CF.  Due to the high degree of variability of this effect, it does appear unlikely at this time that this approach would have widespread therapeutic benefit.  However we are continuing our studies to get a better understanding of what PAR1 does in the CF lung in case there is still a potential approach we might be able to use.

 

Modelling complex variation in multiple breath washout (MBW) data to reveal unobserved characteristics of early childhood lung disease in cystic fibrosis

Professor John Carlin  

$ 31,520 Grant

Progress Report - November 2016

The MBW technique has been widely adopted by respiratory researchers because of its usefulness in assessing lung function of various common and serious lung diseases, particularly cystic fibrosis.

In this project we have developed and applied sophisticated statistical models and techniques to MBW data, and we expect in continuing work to reveal properties of the data that are clearly connected to the underlying lung biology.  These advances will give researchers new perspectives on early childhood lung function, with a particular focus on identifying early markers of disease progression in CF.

A related goal is the refinement of MBW and similar techniques to make them suitable for clinical use. Improving the statistical methodology have taken us closer to this goal and we expect our fundings from analysis of research data to demonstrate the clinical utility of these techniques.  This research should make MBW testing more useful and cost-effective for both research and clinical purposes, thus providing a powerful tool for the study and management of CF.

We believe that we have generated new data and an interesting mathematical analysis that will be of interest to the CF community as new treatment trails use the MBW test.  Our method means even when the testing time is short, as is often the case in young children, or relatively short as it may be in those who are sicker, useable results will be available in more children.

Role of an acute phase protein Alpha 1-acid glycoprotein as a biomarker of prediction of pulmonary exacerbation in young patients with cystic fibrosis

Rosemary Carzino - Progress Report

$ 41,367 Grant

Progress Report - November 2016

This project aims to investigate the role of an acute phase protein in blood as a biomarker for the prediction of a pulmonary exacerbation in young patients with CF.

To date we have not yet analysed any data for this project as the samples are currently being tested and results are yet to be received.  Upon discussion, we have decided to change direction in methodology and send the samples off to a biomedical company in Finland, called Brainshake Ltd.  The advantage being, besides analysing for the acute phase protein levels in question, a further 200 other biomarkers can also be tested simultaneously in a small volume of one sample by the use Nuclear magnetic Resonance technology.  This also allows for a great opportunity to work alongside other groups in MCRI who are also utilising this service where results may be compared for further analysis.

Once results are received from Brainshake, they will be analysed alongside relative clinical information to ascertain the ability of the acute phase blood protein to predict a pulmonary exacerbation in young children with CF, with the added benefit of exploring other blood biomarkers in the meantime

2014 - MCRI

feasibility of human Epithelial Cell culture from Bronchial Brushings of cystic fibrosis patients for studies of airway modelling and research.

Paul Griffin, Cilial Function Scientist

 $15,000 grant

In this project we will use samples obtained from bronchial brushings obtained during broncho-alveolar lavage to establish a novel model and validate an in vitro model. This model can then be used to study a variety of conditions, including pathogenesis of, and recovery from various infectious organisms and the effects of environmental physiological changes on the respiratory epithelium. This will be done by growing the cells in an air-liquid interface culture using antibiotic media. The antibiotic clears infections in the sample allowing the cells to re-differentiate into “clean” new, airway epithelia. The cells are then challenged under different conditions and a comparison can be made to normal airway epithelia. Changes in the gross structure, function and ultrastructure of the cells will be analysed using high speed video and electron microscopy (and possibly fluorescence microscopy).The model can further be used to test a wide range of pathogens in addition monitoring the time it takes to completely recover to normal function and the influence of pH on epithelial and ciliary function..

 

The aim of this project is to adapt the cell culturing technique currently used on nasal epithelial brushings for bronchoscopic brushings from CF patients. Once validated, a functional and biologically representative cell model can be used to perform experiments. If successful, a second aim of this project would be to investigate the pathogenic sequence of infection of bacteria commonly isolated from CF specimens. The model can be used to check the time of clearance of the bacteria using antibiotics and how long recovery of normal function takes. Also, the effect on important aspects of the airway including cilia beat frequency and differences in ultrastructural components can be monitored. Thirdly, environmental conditions such as the pH of the media and Sodium Bicarbonate levels can be varied and again, the effects on functions and structure of the cells can be observed.

 

Very little detailed work has been reported on primary, differentiated CF airway epithelial cell culture. Most studies are done on explants or expensive commercial cell lines which are not as relevant. This project will provide an improved cell culture system for respiratory research that can provide novel experimental investigations. It also has the potential to allow for individual reports on patient samples.  Our wealth of experience in cell culture, with the addition of world leading equipment and support (currently utilised in the RCH PCD diagnostic service), have shown images and videos produced here, are of the highest quality. Also, as it is thought that this method of culture should be more successful, it would therefore in theory, reduce the expense and time spent on culturing samples that fail to thrive. A success model would be attractive to scientists and clinicians studying mucosal immunology, CF airway therapeutics and a myriad of other disciplines. To our knowledge, less than a handful of centres in the world possess this model for studying young children with CF.

 

Evaluating the role of elastase in the inflammatory response in cystic fibrosis

Associate Professor Phil Sutton, Group Leader, Mucosal Immunology

$45,000 Grant

Cystic fibrosis (CF) patients are highly susceptible to respiratory infections that result in life-threatening inflammation in their lungs. A/Prof Ranganathan has identified that a protease (an enzyme that cuts up proteins) called neutrophil elastase is significantly associated with CF pathogenesis. This elastase is probably produced in response to infection of the lung.

  This project builds on a finding of Investigator Ranganathan of potentially major significance that was published in one of the top medical journals.

 Our findings indicate that the detection of free neutrophil elastase in lower respiratory secretions is associated with the development of early bronchiectasis in CF. The aim of this project is to investigate the potential use of this observation to identify suitable new targets for therapy in CF. This proposal will provide a proof-of-concept test of a theory regarding how we believe neutrophil elastase contributes to bronchiectasis in the airways. If correct this will identify new ways to prevent the pathway to destructive lung damage.

 An important additional significant bonus of this project is that it will establish the CF mouse model at MCRI. Its introduction will increase the variety of techniques available for CF research.

2013 - MCRI

exploring the early lung microbiota in cystic fibrosis

New techniques have revealed that a much larger range of organisms than we previously believed colonise the human lung. These microbes are called the microbiota. We don't know which organisms reside in the lungs of people with CF. We believe it is important to study this as we have identified that the lungs can be damaged in CF before typical infections are detected. We hope that by turning our attention to different microbes that might be present in the lungs early in life and that may initiate the inflammation that causes damage to the structure of the lung.

 The Royal Children's Hospital and MCRI have been at the forefront of studies into early lung disease in CF for more than two decades. Previously (1992-1995), we stored a unique resource of lower respiratory and saliva samples from infants and young children with CF, together with samples from controls without CF who underwent lung washings for investigation of other diseases. Together with our leading team of scientists from the Human Microbiome Project in the US, we now want to analyse these samples in order to explore the range of microbes in early CF. This will enable us to investigate several important questions about the potential role of the microbiota in the development of CF lung disease. 

Project update (January 2015)

1)      Microbial diversity in lung fluid

Our analysis has now revealed the relative abundance of genera identified in the historic BAL samples from children with CF treated at RCH using 16s PCR sequencing. Deep sequencing is ongoing and will be used to identify individual bacterial species, virulence patterns and antibiotic resistance genes. Dr Katherine Frayman will be looking at long term clinical outcomes in relation to these early data. We are currently exploring how these sequencing data relate to conventional microbial culture and other aspects of disease pathogenesis and important clinical outcomes in preparation for publication. We will be the first group able to report on the microbiota in relation to inflammation and other clinical outcomes and so a manuscript is in preparation.

2)      Differences between the left and right lung

We have assessed the number of reads and the relative abundance respectively of microbial genera in lung fluid samples obtained from the left and right lung. Where there were sufficient reads using 16s sequencing on BAL samples obtained from the left and right lung we identified that microbial genera were similar in most cases. In one case, there was a stark difference between left and right lung: the genus Staphylococcus was identified in the right lung but diverse flora in the left. We are investigating pulmonary inflammatory markers and evidence for regional structural lung damage in these individuals. This is a unique appraoch as few centres have recourse to such samples from the left and right lung.

3)      Changes in the microbiota over time

In fifteen subjects at least three BAL samples were analysed successfully so that changes in the microbiome over time can be analysed. . Again, the data are unique because we are able to compare the microbiome with changes that occurred in pulmonary inflammation, standard culture and clinical progress. We are writing up the results and believe they will be of great interest to the CF and respiratory research community in helping to better understand the microbiota in the CF lung.

 

Prevalence of small-colony variant s.aureus among children with CF

 

Lung infections with the microorganism, Staphylococcus aureus, are common in young children with cystic fibrosis (CF). Many patients experience repeated infections during which time S. aureus adapts itself, becoming better suited to living in the human lung by forming what are known as small-colony variants (SCVs).  

These changes make the bacteria more resistant against treatment with antibiotics and affect their ability to grow in the methods laboratories currently use to diagnose bacterial infections. Because this slower growth makes them more difficult to detect, at present we do not know whether SCVs in S. aureus are an important cause of infection in our patients.  

This study will use updated diagnostic methods to specifically look for SCVs in respiratory specimens collected from patients attending the Royal Children’s Hospital (RCH) Melbourne, and we expect that SCV infections will be common in children with CF lung disease. Detection of these variants is particularly important, given their ability to resist treatment with certain antibiotics. Knowing the prevalence of this bacteria will help us to improve the way in which we diagnose respiratory infections in CF, and will also provide clinicians with more detailed information that can be used to optimise the treatment and management of their patients.

 

Project update (January 2015)

 

Twenty six children (mean age 11.35(5.05) years) from the AREST CF study (Melbourne) were identified with a positive S.aureus culture from lower respiratory tract specimens collected in 2014. All children were on at least one or multiple antibiotics prior to or at time of collection. The antibiotics were, namely Augmentin Duo (46%), Tobramycin (15%), Bactrim (11%), or other (27%). Fifty-seven percent of samples were visible on large MSA plates and subcultured onto wild-type HBA plates, but no SCVs of S.aureus were isolated.  

Conclusion: SCV’s were not isolated from the lower airways specimens and therefore were not identified as contributing to lung infections in our cohort of young subjects with CF.

 

establishing predictive indicators of amino glycoside nephrotoxicity and the incidence of toxicity in children aged less than 6 years with CF

 

Cystic fibrosis (CF) is caused by a gene defect and affects the lungs and other body systems. People with CF have altered medicine metabolism. This is because CF affects how the body processes medicines.  

Aminoglycosides are antibiotics commonly used in the treatment of CF. As CF is now mainly diagnosed by newborn screening, aminoglycosides are being used earlier than in the past. Thus people with CF are exposed to higher doses and to longer and repeated courses. This long and early exposure to aminoglycosides may pose risks that little is known about appropriate dosing or toxicity. 

There are two important side effects from aminoglycosides. These are:

 - kidney damage and;

- hearing loss or deafness. 

The aim of this research project is to see if we can discover markers that show changes in kidney function, before and after treatment with aminoglycoside antibiotics. We will do this by testing blood and urine samples.

 The research project may help to find a marker of early kidney disease. This would help us to know when to reduce the dose of aminoglycoside or when to change the antibiotic to a different one, to prevent damage to the kidneys.

project update (January 2015)

 

Patients under six years of age were recruited when commencing IV aminoglycosides (gentamcin 7.5 mg/kg or tobramycin 12 mg/kg) and had urinary and serum sampling of creatinine and magnesium.

A total of seventeen patients aged 0.74-6.99 years, ten males, eight of them on gentamicin and nine of them on tobramycin, have been recruited to date. A total of 44 patients will be recruited.  

Early results suggest that the metabolism and excretion of magnesium in CF warrants further study, and that aminoglycosides (gentamicin>tobramycin) considerably alter magnesium excretion.

Thank you to all 65km for Cystic Fibrosis participants and their supporters for generously supporting our research projects