Summaries of 2016 research

What animals are you planning to use?

Immunodeficient and immunocompetent in-bred strains of mice. Genetic alteration of mice will take place in a minority.

For what purpose are the animals going to be used?

Our animals will be part of a study to investigate the causes, and potential cures for metastatic cancer. A minority of the animals will be genetically altered to establish how specific mutations lead to cancer. The majority of mice will be transplanted with tumour cells in order to study how cancer cells grow and spread around the body, and to evaluate new therapeutic strategies to prevent metastasis, and to eliminate cancer cells.

What will be the harms to those animals and how will these be limited?

Mice will develop tumours and these may spread to multiple organs. We monitor these animals daily for signs of stress and discomfort and if this cannot be controlled the animals are humanely killed. A proportion of the animals will carry tumours that can be readily scanned, which allows us to detect the tumours at their very earliest stages before the mice are likely to be adversely affected by the disease.

What alternatives did you consider before embarking on the use of animals in your research?

All of our studies are performed in cell culture where the majority of measurements and experiments are performed. Only when we are sure of the efficacy of a particular intervention or the function of a specific gene in cancer, do we confirm these findings in animal models. Hence the animal models are used sparingly as a final confirmation of our in vitro studies.

What will be the expected benefits?

These animal studies are the final step in either the identification of a new, previously undiscovered, mechanism of cancer, or in the development of a novel therapeutic intervention. Thus, this work directly contributes to the advancement of knowledge of cancer, and in the development of new anti-cancer agents for the clinic.

What animals are you planning to use?

The animals mainly used in this study will be mice. As we are studying brain development and behaviour, our animals will range from embryonic to adult. Some of these mice will be genetically altered to induce decreases or increases in the activity of certain genes. The animals used in this study are already in existence and will not be generated in Cardiff. We will also be using some rats in our studies of how the environment may moderate gene activity.

For what purpose are the animals going to be used?

Our genes encode a range of molecules that are essential for development and normal function. We know that in addition to what these genes encode, when they are active and how much product they produce, is also critical. Our animals are part of a study to understand the effects of changes in the levels of gene activity on how the brain develops and whether this results in altered behaviour. This process often goes wrong in neurodevelopmental disorders, and we want to model that. We also want to study how the environment experienced by a developing fetus in pregnancy can also affect gene activity and brain development.

What will be the harms to those animals and how will these be limited?

The main harms may arise from how we perform the behavioural testing of the animals. However, we have considerable experience of behavioural experiments with rodents and have fine-tuned the tests to minimise any unnecessary stress and anxiety. This is in our interests as well as in the animal’s interests, as anxious or stressed animals produce poor behavioural data.

What alternatives did you consider before embarking on the use of animals in your research?

It is currently not possible to model the environment of pregnancy in a cell-based system. Nor is it possible to effectively examine behaviour other than in a living animal. However, where possible we will be using cell models to examine brain development.

What will be the expected benefits?

Our project will provide us with a better understanding of how brain and behaviour is affected by changes in gene activity, something that is a known risk factor in a number of neurodevelopmental disorders. Our research will also identify environmental factors, such as diet during pregnancy, that may alter gene activity and predispose the offspring to neurodevelopmental disorders.

What animals are you planning to use?

We will be using adult mice, mostly wildtype, but some also genetically altered. The genetically altered mice are obtained from other laboratories, and will not be generated at Cardiff University.

For what purpose are the animals going to be used?

We are conducting a study on brain cancer to understand why these cancers are so aggressive and to develop new therapies for patients. As part of the study, animal research will help us to better understand the interactions between cancer cells and normal cells surrounding them.

What will be the harms to those animals and how will these be limited?

The animals will grow brain cancers, but as the brain has no pain receptors this process is in and of itself pain-free. In order to implant some of these cancers, some animals will undergo surgery, and we will use neurosurgical techniques, anaesthesia and medication (very much in the same way as neurosurgeons operate on human patients) to minimise any harm to the animals.

What alternatives did you consider before embarking on the use of animals in your research?

We have considered in vitro models of brain cancer, but these models cannot recapture the complexity of both the tumour and the normal cells surrounding it. It is becoming increasingly evident that the normal tissue around a cancer is not just sitting there, but both reacting to the cancer and also involuntarily contributing to its growth. Nevertheless, we always study brain cancer cells in the lab (in vitro) first before continuing our observations in an animal. We are also striving to improve currently existing in vitro systems by developing better models where cancer cells and normal cells are cultured together and we will use these systems to further refine our research.

What will be the expected benefits?

We expect to improve understanding of the biology of brain cancer, which will help in developing new and better therapies for patients. In the past, a large proportion of new drugs developed in the laboratory have failed to work in patients, and this is at least in part due to the fact that isolated cultures of cancer cells in vitro are not representative of the tumour inside a patient. Several types of brain cancer are currently incurable, and we expect that by studying the cancer in its appropriate environment – the brain – we will be able to identify new ways of how these cancers can be treated.

What animals are you planning to use?

We will use adult mice, some of which will be genetically modified. These mice will be obtained from other academic Institutions or commercial sources.

For what purpose are the animals going to be used?

The animals will be used to understand how the immune system recognises cancer and whether it can be boosted to recognise and kill cancer more effectively. During the study, animals will, for example, be given novel vaccinations to find out whether these can induce immune responses that are capable of destroying tumours. In other cases, they will be given new drugs to test whether these can help activate the immune system for the same purpose. Our findings have already shown us that in the case where strategies aimed at inducing immune responses work, they work only in a proportion of animals. With this in mind, we will also work out what the differences are between animals that respond to treatment (where the immune system kills the cancer) and those that do not (cancers carry on growing).

What will be the harms to those animals and how will these be limited?

Animals will be handled and restrained in order to deliver e.g. vaccinations or drugs and for the purpose of monitoring tumour growth. Suffering is minimised by excellent handling techniques which limit contact time. There is strict adherence to tumour monitoring procedures which ensure that tumours cause no distress to the animal.

What alternatives did you consider before embarking on the use of animals in your research?

It is clear that the immune system can under some circumstances clear cancer but unfortunately, the precise details of those circumstances remain poorly understood. The immune system consists of a large number of different cell types which perform different functions in different parts of the body. This complexity cannot yet be recapitulated in the test tube therefore animal models remain necessary for examining the function of a fully integrated immune system. New methods however are being established and used to significantly limit the number of animals required to address key questions. We are developing the use of three dimensional organoid systems, which enable us to generate cancer in the lab instead of in animals. These well-characterised entities, can be implanted into animals, allow us to address questions relating to the immune system using fewer experimental mice. In addition, we use a variety of imaging systems to scan individual mice refining the accuracy of tumour measurements allowing conclusions to be drawn on smaller number of animals. Furthermore, use of these techniques will facilitate serial longitudinal analyses of tumour growth in individual mice, contributing significantly to reducing the number of animals required to achieve the study objectives.

What will be the expected benefits?

Our laboratory performs observational studies in patients with cancer and tests hypotheses arising from findings of these studies in preclinical mouse models. New information gained in this way is used to inform early phase trials in patients with cancer. We have a track record in this area thus the mouse to man pipeline is well established.

What animals are you planning to use?

The research uses adult rodents (rats and mice). While the overwhelming majority of rodents will be wildtype, in a few instances the rodents may be genetically altered to examine the impact of genes associated with and increased risk of disorders such as schizophrenia. These rodents will be imported to Cardiff University.

For what purpose are the animals going to be used?

Learning and memory depends on the interactions of multiple structures within the brain. The same structures that are thought to be vital for human memory are found within the rodent brain. We are seeking to understand how these structures are interconnected and how they make different, but related, contributions to memory. For this reason, the studies involve anatomical and behavioural techniques, the latter to examine different forms of memory. Starting with evidence from human amnesia, we can narrow down and identify the candidate structures that are necessary for different types of learning. Our interests are in how these structures co-operate normally and how these structures respond when one of them is damaged, as potentially happens in various clinical conditions. The focus is principally on structures within two brain regions, called the temporal lobe and the diencephalon.

What will be the harms to those animals and how will these be limited?

To measure how brain regions normally interact during learning we can measure brain changes associated with forming memories. One example concerns the production of proteins by the brain that help to form and stabilise long-term memories. To identify the precise location of these proteins it is necessary to look at brain tissue after the rodent has been humanely killed. In other studies the goal is to determine whether a brain site is needed for a particular aspect of learning. Such studies involve temporarily or permanently disrupting that specific brain site. These studies often involve surgeries, which always occur under general anaesthesia. None of the sites targeted affect sensory or motor function. Because it is vital to assess learning by examining behaviour, it is critical that the animals do not have wider patterns of problems.

What alternatives did you consider before embarking on the use of animals in your research?

The research concerns the normal interactions of an incredibly complex system. An integral component concerns the ways in which different structures work with each other to produce behaviour that demonstrates learning. For these reasons, it is necessary to examine extensive arrays of brain systems. Non-invasive imaging techniques do not have sufficient resolution to identify the separate brain sites that make up the primary focus of this research (nuclei within the anterior thalamus). To limit the use of animals we have devised new, more sensitive behavioural tests. By examining patterns of gene expression associated with learning, we are able to look at the relative contributions of a multitude of brain sites in a single case, so maximising available information.

What will be the expected benefits?

Developing our understanding of areas of the brain that disturb memory in conditions such as Alzheimer’s disease. Research into the brain’s systems for memory has consistently focussed on a structure called the hippocampus. Our research has revealed the parallel importance of another site, the anterior thalamic nuclei. These same animal findings prompt non-invasive, imaging studies in humans, which seek to focus on these same regions and their interconnections.

What animals are you planning to use?

We will be using mice, some of which will be genetically modified to change their predisposition to cancer. We will import some strains of mice and also generate some strains at Cardiff.

For what purpose are the animals going to be used?

Our animals are part of a study to help us understand the basic mechanisms underlying cancer development and progression, to identify novel therapeutic targets and to help determine the usefulness of new anti-cancer therapies.

What will be the harms to those animals and how will these be limited?

The adverse effects will be the development of cancers and the negative side effects of anti-cancer therapies. Our mice will be monitored for these adverse effects and humanely killed if multiple moderate adverse effects are seen.

What alternatives did you consider before embarking on the use of animals in your research?

We are increasingly using alternative models of cancer, most notably human tumours maintained in tissue culture. Whilst these cannot yet fully model tumour development in a whole organism, we are continuing to refine this approach such that it could ultimately replace our animal models.

What will be the expected benefits?

The knowledge we will derive will underpin future strategies for the development and use of novel anti-cancer therapies. This is particularly relevant to the treatment of cancer, as at present there are relatively few new options available in the clinic to modulate the course of disease. The majority of clinical benefits are likely to be long term; our principal hope is that by identifying candidate genes and candidate genetic pathways we will be able to refine and accelerate drug development for human therapy. There will also be short term benefits, primarily from the testing of novel drug therapies within our models. This latter approach has the potential to directly and immediately modify clinical practice.

What animals are you planning to use?

In our research we use South African clawed frogs (mainly Xenopus laevis and to a lesser degree Xenopus tropicalis) as they are the best model for developmental studies of this type.

For what purpose are the animals going to be used?

Our animals are used to provide embryos, which are required for our research on heart development.

What will be the harms to those animals and how will these be limited?

To induce egg laying (and mating behaviour in males), injection of pregnancy hormone (chorionic gonadotrophin) is used, and this procedure can on rare occasions cause skin irritation or superficial wounds. The toads we use are exceptionally robust and fully recover from any such injury.

What alternatives did you consider before embarking on the use of animals in your research?

Mammalian pluripotent cells (embryonic and induced pluripotency state cells. ESC/iPSC) have a recognised translational potential for cardiac regenerative medicine and disease modelling in vitro. In order to realise this potential in full, it is necessary to improve our knowledge of how the heart and its components develop by studying heart development in vertebrate models such as Xenopus. We use both complementary models- mammalian ESC/iPSCs and Xenopus embryos in our research. As the knowledge of heart development improves, it leads to a gradual shift in use of in vitro models and concomitant reduction in reliance on in vivo models.

What will be the expected benefits?

The main benefit is in the form of improved understanding of heart development. Like most aspects of embryonic development, heart development is common to all vertebrates, and what we learn using Xenopus embryos is largely applicable to humans.

There are two main areas of potential applications of this kind of research: heart failure and congenital heart disease. Heart failure results in a loss of working heart muscle and the best treatment for it would consist of a timely replacement with new heart muscle. The main role of research on heart development here is to contribute to strategies for efficient generation of heart muscle tissue: if we learn how an embryo makes heart muscle, we might be able to apply that knowledge in a clinical setting. Congenital heart disease is the most common form of birth defects, with often poorly understood causes. Research into heart development can provide better insight into the underlying causes, and perhaps also offer strategies for development of new treatments.

What animals are you planning to use?

Adult wild type rats.

For what purpose are the animals going to be used?

Parkinson’s disease is a slow degenerative disease that affects over 120,000 people in the UK. The gradual death of a specific set of cells in the brain leads to slowness of movement, tremor and stiffness. Current treatment options are limited as the disease advances, and new therapies are being explored to repair the damage caused to the brain by using altered viruses or cells implanted in to the brain. In order to assess the safety and efficacy of these new approaches we need a good representation of the disease in an animal model. This project will involve animals as models of Parkinson’s disease and attempt to recreate these models as close to the patient situation as possible to test these new therapies.

What will be the harms to those animals and how will these be limited?

The majority will undergo 2 short surgical procedures, firstly to produce the Parkinson’s disease on one side of the brain and secondly to repair the damage. They may be anaesthetised for brain imaging studies but the process of using these studies reduces the number of animals that are needed overall. The rats are given pain relief after surgery and are monitored closely throughout the experiments. Parkinson’s patients receive daily medications and the same is replicated for short periods of time in the rats. This is delivered typically by injection and the place of injection is changed daily to ensure that the animals do not become sore.

What alternatives did you consider before embarking on the use of animals in your research?There are no real alternatives to this type of research. The very nature of the research is that we are trying to replicate, as close as physically possible, the environment in which a new therapy might be trialled in patients so that we may anticipate or prevent any potential problems. To do this we need a whole system, and a cell culture cannot reproduce this.

What will be the expected benefits?

The overarching aim of this work is to optimise new treatments for Parkinson’s disease and ensure that these new therapies are safe and compatible with a patients’ existing medications.

What animals are you planning to use?

We will use mice, both juvenile and adult. The majority will be genetically altered, bred from imported lines.

For what purpose are the animals going to be used?

The animals are part of a study to help us understand how the visual areas of the brain adapt to visual experience made during childhood, why this can sometimes lead to vision disorders such as lazy eye, and how different brain areas work together to produce visual perception.

What alternatives did you consider before embarking on the use of animals in your research?

We have considered in vitro (‘in the dish’) which can be useful in elucidating certain cellular aspects of brain function but it is impossible to maintain a whole brain and eyes alive in isolation in order to truly test brain function, let alone behaviour.

We are using computer-based modelling which has helped us in the past to interpret results obtained in vivo, but data collected from animals are needed to feed into any models to ensure they have a sound basis.

What will be the harms to those animals and how will these be limited?

Animals will be subject to high-resolution brain imaging for which they need to be prepared surgically under general anaesthesia. There is a risk of post-operative pain as well as of infection; these will be minimised by treatment with analgesics, antibiotics and anti-inflammatories as appropriate.

What will be the expected benefits?

We hope to find out, at the level of individual cells and molecules, how we learn to see and what goes wrong in conditions such as lazy eye but also in neurological disorders (such as autism or schizophrenia) that can affect sensory systems. If we discover the mechanisms leading to those disorders then we will have a better chance of developing new methods of treatment.

What animals are you planning to use?

We will use young and adult mice that are both wildtype and genetically altered. The mice are already genetically modified and none will be performed at Cardiff University.

For what purpose are the animals going to be used?

The animals are part of a study to help us understand why people get type 1 diabetes. In addition, the mice, like humans, get diabetes very similar to the human disease, if they are not treated. They will be involved in research to find ways of preventing and curing diabetes.

What will be the harms to those animals and how will these be limited?

Most of the animals will not experience any harmful effects. Some animals may experience minor discomfort for a few seconds when an injection is given. Some animals will develop diabetes, where they will become thirsty and pass more urine. They may start to lose weight. We will either treat them with insulin to control the diabetes, or they will be humanely killed before they have significant illness or weight loss.

What alternatives did you consider before embarking on the use of animals in your research?

The diabetes studies will be combined with tests in cell culture and where possible, the research will be done in cell culture

What will be the expected benefits?

Benefits from this project include better understanding of the development of type 1 diabetes as the model that we use has some very similar features to the human disease. The mice will be involved in research to prevent and cure diabetes using a number of different strategies. If these are successful, the aim is to take this in the next steps towards applying these strategies to human diabetes.

What animals are you planning to use?

We will use mainly embryos and young animals from wild type and genetically altered or GA mice. The GA animals are purchased from commercial suppliers or obtained through collaborations with other laboratories.

For what purpose are the animals going to be used?

Our animals are part of a study to help us understand how a specific type of proteins, called delta protocadherins, contribute to the development of the cerebral cortex. These proteins are located on the membrane of the cells to help them recognize other cells and interact with them. One of the delta protocadherins is mutated in a human disorder called Juberg-Hellman Syndrome, in which very young girls develop epilepsy and other neurological symptoms. Why this happens is currently unknown, and our animals will help us to investigate what goes wrong in the brain of these girls and develop targeted therapies to help them.

What will be the harms to those animals and how will these be limited?

The procedures that will be used are not expected to cause more than temporary discomfort to the animals. Many of them will be humanely killed to obtain tissue for our research and some of them will receive an injection before that. Other animals will undergo surgery. This is conducted under anaesthesia, following aseptic technique to ensure the maximum level of safety, and always using analgesia.

What alternatives did you consider before embarking on the use of animals in your research?

As the cerebral cortex develops, it receives cells from different parts of the brain and signals from varied sources. These cells and signals influence the processes that are necessary for the cortex to form: the generation of neurons, their migration to their final position within the cortex and the contacts that they establish, with other neurons in the cortex or in other brain regions. Because of this complexity, it has not been possible yet to create an in vitro model to study cortical development. However, we also use cell culture when we study basic mechanisms that do not depend so much on the complex arrangement of cells and signals of the in vivo cortex.

What will be the expected benefits?

Our research will generate new knowledge about the function of the delta-protocadherins during cortical development, contributing to our understanding of brain development and function. This is a very important field, considering the heavy burden that psychiatric and neurological diseases pose not only on society, but also on the health systems of many countries. In addition, we expect to shed light on the pathophysiology of Juberg-Hellman syndrome. We will then be able to provide information to affected families to help them understand the disease, but also to clinicians, to guide them in the development of new treatments.

What animals are you planning to use?

Adult wild type rats.

For what purpose are the animals going to be used?

Our animals will be used in a study to determine the changes that occur in the retina in glaucoma. Glaucoma remains one of the commonest causes of vision loss in which the retinal cells that send light signals to the brain die, resulting in loss of vision. We will undertake research to see if it is possible to enhance the remodelling and healing responses of these cells to recover vision that has been lost in glaucoma. If successful, our work will, for the first time, lead to clinical treatments to recover some of the vision that has been lost in this disease. Our work is essential for the design of clinical trials to reverse vision loss in glaucoma patients.

What alternatives did you consider before embarking on the use of animals in your research?

We did consider the use of culture systems run at elevated pressures since this can simulate some of the direct pressure effects on neuronal tissue. However we rejected this since it does not emulate the appropriate neuroglial/ immunological changes that occur in vivo in glaucoma.

We also considered using the mouse as a glaucoma model but decided against it as the mouse eye is small and difficult to assess in vivo and the rat will provide a much better and robust inducible model of glaucoma.

We do use an in vitro model of retinal ganglion cell degeneration based on the retinal explant. This is ideal for studying the effect of therapeutic agents and for determining any adverse interactions between agents. The explant model is based on retina harvested from animals which have been humanely killed.

What will be the harms to those animals and how will these be limited?

The harm level is mild since all procedures are conducted under general anaesthetic. As part of our research we generate chronic experimental glaucoma in rats, but just as in humans, this does not cause any discomfort related to the eye.

What will be the expected benefits?

Our experiments will provide a proof of concept that it is possible to remodel and rewire the retina to reverse the loss of vision in glaucoma. We do not expect to achieve total reversal but partial recovery would still be clinically significant and a great benefit for patients. We hope that treatment for glaucoma will be a single injection into an eye, to allow the parts of the retina that have degenerated to recover and regrow. The therapeutic agent that we will use is already in clinical trial for the recovery of function in spinal cord damage so we are optimistic that our experiments will form the basis for the design of clinical studies.

What animals are you planning to use?

We are planning on using 10-12 week old adult mice, both wild type and mice that are genetically altered (GA) so they are unable to make lipid (fat) processing enzymes. These GA strains are currently bred in Cardiff University.

For what purpose are the animals going to be used?

Our animals are part of a study to understand the role of specific lipids (fats) that are found in the skin during repair. These fats are made by immune cells that travel to the site of a wound after an injury. The role played by these fats at the site of a wound is unknown and we are keen to study if the presence of these fats changes how a wound heals.

What will be the harms to those animals and how will these be limited?

A small punch biopsy will be made to the back of the mice. The wounds will heal within one week and cause little irritation to the mouse. The mice will be monitored continually following biopsy to ensure they suffer no adverse effects as a consequence and pain relief will be administered when necessary.

What alternatives did you consider before embarking on the use of animals in your research?

Within this project we are using a cell culture model. We will make the same small biopsy in this model and add lipids of interest and study the wound repair. However, a drawback of this model is firstly that immune cells, that generate the lipids of interest, are not present, and secondly it is not possible to genetically alter skin cells used in this approach.

What will be the expected benefits?

The primary expected benefit of this research is to assist us in designing possible wound therapies to change the levels of these lipids in human and animal wounds to speed up recovery times. If we find that these lipids aid skin wound recovery we can develop formulations to add into wounds to increase their levels at the site of action. Alternatively if we find they slow healing in any way, we can develop formulations to inhibit their production.