Winter 2008 Newsletter

PhD Student Anne Bielemeier and Dr Lindsay Marshall

New Research into Cystic Fibrosis 

Dr Lindsay Marshall, Aston University

Cystic fibrosis (CF) is the most common inherited life threatening disease in the UK, affecting over 8,000 people.  Data available from the website of the UK’s dedicated CF charity, the Cystic Fibrosis Trust, declare that 5 babies are born with CF every week and three people with CF die.  The average life expectancy is only 31 years, since an accumulation of thick, sticky mucus in the lungs leads to chronic infections which irreversibly damage the airways.  The gene which is faulty in CF was identified 20 years ago and has been the focus of intensive research ever since.  The “cure” for CF would be replacement of this faulty gene with a fully functioning copy and many research groups around the world are concentrating on finding ways to bring this treatment to people with CF.  However, since gene therapy is not yet available, there is still a need for better treatments and, ideally, preventative medicine to stop the lung damage caused by the infections.

The aims of our research project are simple, achievable and of vital importance in the quest to develop effective treatments for people with CF.  We aim to develop accurate models of human airways in the laboratory to use for testing potential therapeutics.  We also plan to use our cell culture techniques to generate a valid model of the CF airways in the laboratory so that we can mimic the infections typically observed in people with CF.  We will use human cells grown under appropriate conditions which allow the cells to develop the features and, importantly, the functions that they perform normally in people.  Currently, mice artificially manipulated to express the faulty CF gene are used for this type of study but we don’t feel that this is a suitable model, and this is supported by reports that indicate these manipulated mice do not get the same lung disease as people with CF.

The proposed programme of work will use the models of normal and CF airways to understand the dynamics of interactions between the multiple cell types constituting the lung, a very complicated organ system.  These models will be invaluable for investigating the lines of communication that exist between human cells normally and between the human cells and invading bacteria following infection, as these pathways have yet to be defined for the airways.  This will have implications for all infectious and inflammatory conditions of the airways in humans and it is hoped that it will lead to development of a human testing system for therapeutics, saving animals currently tested with novel compounds.  The model system would also be applicable to evaluating potential anti-inflammatory and anti-infective treatments for conditions other than CF, such as chronic obstructive pulmonary disease (COPD) and other chronic and acute lung diseases.

In CF, there is a characteristic sequence of infections that begins with Staphylococcus aureus early in life.  These bacteria seem to pave the way for more aggressive, sustained infection with Pseudomonas aeruginosa.  It is the sustained infection with Pseudomonas bacteria that then wreak havoc with the structure and function of the airways, and the current antibiotic treatments do not appear to completely eradicate these bacteria. In fact, Pseudomonas infections are responsible for 95% of the mortality in young people with CF. 

The quest to generate an animal model for CF lung disease began in 1979 and established animal models have been employed extensively ever since, in the search for effective treatments and to discover the mechanisms behind the lung disease.  The use of these models may have provided the scientific community with some useful data, but they have not fulfilled what was hoped for. Significant differences have been observed between CF mouse models and human CF disease patterns, particularly in terms of demonstrating airways disease progression – an essential feature of any experimental model applied to validate disease processes in humans.  It is entirely appropriate to develop a valid model of human airways in an attempt to move away from the reliance on such animal models.  I hope that the development and validation of human models will encourage others in the field to take this approach.

The Humane Research Trust Laboratory: Supporting the Future

Dr Michael Wormstone, University of East Anglia

The UEA Team

The Humane Research Trust laboratory is a vital element in the past, present and future successes of the Norwich Eye Research Group and has served as a wonderful centre for research and education. We are extremely grateful that THRT has agreed to extend support for THRT laboratory until the end of 2013. The funding received to maintain the laboratory not only provides excellent facilities (including experienced technical support) to conduct our human tissue research, but also greatly aids our ability to secure additional funds from a variety of sources, to enable specific research projects to take place.  THRT laboratory funding is therefore a catalyst that increases our overall level of support and enables our work on human tissue to be carried out in a highly productive manner.

The laboratory’s primary focus is ocular disease, but it is important to emphasise that much of the work has great relevance to related conditions throughout the body. While our initial drive is to identify the mechanisms regulating eye disease we are always aware of potential applications to non-ocular conditions and how technical advances resulting from our work may facilitate other human tissue studies. The main topics of interest at present in the laboratory are the role of Transforming Growth Factor beta (TGFb) in cataract and PCO (a problem that results after cataract surgery); Endoplasmic Reticulum stress (cataract, PCO and Myotonic Dystrophy); inflammatory molecules (Cataract and PCO); signalling of the neural retina (Dr Julie Sanderson and her team); dietary antioxidants (cataract); drug delivery. These areas of investigation have direct application to ocular disorders, but importantly provide general information applicable to many related conditions. For example, TGFb is implicated in skin wound healing through the modification of cells and contractile processes. In our studies we can identify the basis for these events and translate this information to the skin. The current programme is very active giving rise to numerous publications and keeps redefining the boundaries of our knowledge; I believe the continued support for the THRT laboratory will maintain this trend. 

“In order to discover new lands, one must be willing to lose sight of the shore for a very long time” (anon). 

In addition to being an outstanding research centre, THRT laboratory is an excellent teaching resource. Since 1994, an estimated 50 undergraduates have used The Humane Research Trust Laboratory during final year research projects; at current rates a further 36 students will use the laboratory in the next 6 years. Scientific research is becoming increasingly expensive, therefore funding from The Humane Research Trust to support final year research students will greatly facilitate the running of their projects and aid the student experience. The final year research project is a defining period for any student undertaking a scientific degree. In many cases, this is this first true opportunity for a student to see science in action rather than theory. This experience not only provides opportunities to use cutting edge technologies, but also to witness how science is truly implemented to shape our understanding of the healthy human being and the changes associated with disease. Students often enter the laboratory at the start of their project unable to detach themselves from the gospel that is published literature. Within a project we therefore teach students to look beyond this established body of information toward the unknown. Once the student can take this bold step they actually start to become a true scientist rather than a scientific scholar. In my own experience, I convey to the students that within their project they will be privileged to discover and consequently know something that most likely no other person on earth knows. This is one of the joys of being a scientist and drives you constantly in to the unknown. Supervising final year research students is one of the most rewarding experiences in teaching. This period can have a marked affect on the development of the individual and often confirms a desire to follow a career in scientific research. The input of The Trust is therefore invaluable.

A New Model to Study Inflammatory Signals for Barrett's Oesophagus

Dr Mark Williams, University of East Anglia

Miss Natalia Scobioala-Laker

A collaborative research project between the Gastrointestinal Research Laboratory at the University of East Anglia and clinicians at the Norfolk and Norwich University Hospital will develop a new model to study inflammatory signals for Barrett’s Oesophagus.  The oesophagus is more commonly known as the food pipe or gullet and is the tube that carries food from your mouth to your stomach. Over the last 30 years oesophageal cancer has become more prevalent and is now the 9th commonest cancer in the UK.  There are nearly 8000 new cases diagnosed in the UK each year and the prognosis for this disease is very poor with a mortality rate of 80% (only 10% of sufferers survive for 5 years or more).  Risk factors for oesophageal cancer include age, gender (more common in males), smoking, alcohol and a diet low in fruit and vegetables.  Also, people with a condition called Barrett’s oesophagus are up to 125 times more likely to develop oesophageal cancer.  Barrett’s oesophagus is thought to develop following long-term acid indigestion (reflux) from the stomach which gives rise to chronic inflammation.  In the case of Barrett’s sufferers, the lining of the oesophagus heals itself by replacing the damaged multi-layered lining of the lower oesophagus with a mass of disorganised tubular structures called crypts.  It is therefore imperative that the molecular and cellular basis by which inflammatory mediators promote Barrett’s oesophagus and progression to cancer is understood better in order that preventative measures can be deployed following diagnosis at endoscopy.  Hitherto, the study of Barrett’s oesophagus has been hampered by the lack of an ex vivo human tissue model of the disease. The Humane Research Trust is funding Miss Natalia Scobioala-Laker in the Gastrointestinal Laboratory to conduct a three year PhD programme of research during which time she will place human tissue samples of Barrett’s oesophagus into a novel three-dimensional tissue-culture system.  Natalia will apply state-of-the-art fluorescence imaging technologies to identify the inflammatory factors that promote propagation of Barrett’s oesophagus and progression to oesophageal cancer.

The very first images of a human Barrett’s crypt maintained in a novel three-dimensional culture system.  Cell nuclei are labelled red and proliferating cells are labelled yellow.

Towards a non-animal model or testing new drugs against parasites

Dr David Timson, Queen’s University Belfast

Parasitic worm infections affect millions of humans and farm animals worldwide. One parasite of particular importance is the liver fluke, which infects an estimated 17 million humans, mostly in the developing world and causes billions of pounds worth of economic losses within the global agricultural industry. Infection with liver fluke results in pain, poor growth and, in some cases, death. Treating these infections is therefore a public health issue in the developing world and is crucial in the improvement of both agriculture and animal welfare.

PhD Student Mark O'Shea and Dr David Timson

The current preferred treatment against this parasite is a drug called triclabendazole. This is a safe and effective drug that can be used in both humans and animals. Unfortunately, just as bacteria such as MRSA have become resistant to antibiotics, there are now widespread reports of liver flukes displaying resistance to triclabendazole. Therefore there is now an urgent need for the development of new treatments to combat the liver fluke parasite.

An effective treatment must be successful in killing the parasite while not harming the mammalian host. Current methods used for investigating the action of anti-fluke drugs involve the parasites’ exposure to the drug and observing the responses. Unfortunately the only known way to produce live adult flukes is to grow them within a mammalian host, such as a rat, which must then be killed in order to extract the parasite.

Funded by The Humane Research Trust, Mark O’Shea has just commenced a PhD project, with Dr David Timson at Queen’s University Belfast. Mark is researching alternate methods to test certain groups of compounds. He is ideally qualified for this having a BSc in Pharmacology and an MSc in Molecular Parasitology from the University of Liverpool. Mark will identify the genes coding for calcium channel proteins. These are an important group of proteins as they regulate and control the entry and movement of calcium ions with the cell. Generally, if they go wrong, the cell will die therefore providing an attractive target for a wide variety of compounds. Once Mark has identified the genes, he will insert them into baker’s yeast (in place of the yeast’s own calcium channel genes). This will provide a model for testing compounds for their effectiveness against liver fluke ion channels.  By subjecting the yeast to a series of biochemical tests for calcium channel activity it should be possible to quantify and rank the potency of the various compounds.  In addition to saving animals, the new method should be cheaper, faster and more quantitative than existing ones.

In the long term it is hoped that it will be possible to construct similar model systems for other ion channels, both from the liver fluke and other parasitic worms.

Conference Report:    FASEB Meeting – Signalling through tetraspanins and other cell surface

PhD Student Rachel Hulme

Rachel Hulme is a Humane Research Trust funded PhD student at the University of Sheffield.

As one of FASEB (Federation of American Societies for Experimental Biology) prestigious summer research conferences, this tetraspanin meeting involved the largest number of international research groups in the field.  This year it was held by Yale University in New Haven, Connecticut and was attended by groups from 12 different countries across 4 continents with academics from the USA, UK, Europe, Asia & Australasia.  Although the largest tetraspanin meeting to occur regularly, its attendance of around 100 delegates allows for a more personal and intense approach to the subject by all involved.  This results in a more informative and mind expanding experience where results and ideas can be discussed ‘one on one’ with experts of the field.

I attended the biannual FASEB meeting with my supervisor Dr Peter Monk, and two other collaborators at the University of Sheffield.  We presented three posters and Dr Peter Monk gave a talk in the ‘Cell Motility & Morphogenesis’ session on the tetraspanin CD63, one of many tetraspanins associated with cancer.

I was fortunate enough to attend the FASEB meeting thanks to a travel bursary from The Humane Research Trust, and presented a poster entitled ‘Defining the Active Sites of Tetraspanins’.  As there were multiple poster sessions scheduled throughout the week I was able to meet many researchers, who were very interested in the use of human monocytes and osteoclasts taken from consenting volunteers rather than the use of animals or cell lines.  The poster sessions gave a number of opportunities for discussion about our work and my personal highlight was towards the end of the week when an important contributor to the cell fusion field, Professor A. Vignery from Yale University, took time to meet with me for an hour or two to discuss the work we are currently doing at Sheffield and the very relevant use of human cells.  From all the meetings throughout the week I returned with a number of new ideas and improved direction.

As a very intense meeting over six days, all from Sheffield who attended found it extremely rewarding and informative.  The question sessions following the talks were particularly useful allowing researchers to defend their work and identify areas which could use further experiments in response to constructive criticism.  I think we all returned from the meeting brimming with ideas and contributions to make to other members of the group who were unable to attend, excited about the prospect of continuing with our work.  My thanks again go to The Humane Research Trust for making my attendance possible and their continued interest in and support of my work.

(Tetraspanins are complex proteins on the surfaces of cells involved in many important processes and pathologies.)

Home Office Statistics of Scientific Procedures on Living Animals 2007

Just over 3.2 million scientific procedures were started in 2007, a rise of about 189,500 (6%) on 2006.  The increased animal use was mainly due to increases in the use of mice, fish and domestic fowl, whilst the use of most other species was down when compared to 2006.

Genetically modified animals were used in 1.15 million regulated procedures up 114,400 (11%) representing thirty-six percent of all procedures for 2007, compared with thirty-four percent in 2006 and eight percent in 1995.

Non-toxicological procedures accounted for about eighty-seven percent of the procedures started in 2007.  This contrasts with seventy-five percent of such procedures in 1995.  The main areas of use were for immunological studies, pharmaceutical research and development, cancer research, anatomy and physiology.

Procedures for toxicological purposes accounted for thirteen percent of all procedures started in 2007.  Of all the toxicological procedures conducted in 2007, eighty-seven percent were performed to conform to legal or regulatory requirements.

Crown copyright 2008