Research in to Brain Tumours

 Introducing Alison Howarth

In February 2016 Alison joined the Therapeutics group, part of the Brain Tumour Research Group at Portsmouth, as a PhD student. She will be studying the treatment of paediatric glioblastoma and diffuse pontline glioma, using re-purposed drugs in combination with P13K inhibitors.

Where did your passion come from to research brain tumours?

Before starting my PhD at Portsmouth I was a research scientist for a number of years. It was during my degree, and continued, when I was completing my masters that I became very interested in research into orphan, or neglected diseases. These are diseases which are, for a number of different reasons, under-funded and under-researched. I knew that I wanted to be a part of the research into these types of disease, and that I wanted to go into drug development.

Soon after my masters I started work as a research scientist at Pfizer Pharmaceuticals, working in high throughput screening labs testing large drug sets to try to find new ways of treating diseases.

After working at Pfizer for a number of years I started work as a research scientist at the University of Oxford where I was working with the Oncology group to find new therapeutics to treat cancer.

It was while working here that I started to work on new ways of targeting brain tumours and I absolutely knew that I wanted to be a part of this research.

Brain tumour research is an area I am particularly interested in because there is an urgent need to find new therapies for children with brain tumours. Currently, the treatments which are available are not always effective, and their use can lead to life changing side effects for children.

These types of brain tumour are considered rare – when compared to other types of cancer – and therefore receive only a small percentage of the national cancer research spend. This means that the advances in treatments for other types of cancer have not been seen in brain tumours.

I have been interested in drug repurposing for some time. Using drugs which are already used to treat other diseases is a good way of using something to its full potential. Using a drug to treat brain tumours which is already used to treat, for example depression, means that the drug can potentially get to the clinic much quicker and cheaper, than discovering or validating a totally new molecule. This is of particular importance for paediatric high grade glioma, where there is an urgent need and where spending may be limited.

The brain is also very interesting to me, it is different to any other part of the body – it is beautiful and complex and the research I am doing is both interesting and rewarding.

 

What are you researching that is funded by the Ollie Young Foundation?

My project is focused on researching new or more effective ways of treating brain tumours in children. More specifically, I am researching potential new therapies for paediatric high grade glioma – types of brain tumours which arise from specific cells in the brain. These tumours are very aggressive and difficult to treat because the tumours are often very resistant, they occur in areas of the brain involved in critical functions, and may have severe side effects.

There is a critical need for new therapies for treating these types of brain tumours in children. In the past, paediatric high grade gliomas have been assumed to be the same as adult high grade gliomas and have therefore been treated with similar drugs and treatment approaches.

We now know that adult and paediatric high grade gliomas are very different to each other; the biology of children’s brains is different to an adult’s, their brains are still developing so treatments which are used in adult patients may have severe or long lasting side effects in children.

Paediatric high grade gliomas are also biologically very different to adult tumours; they occur in different parts of the brain and are genetically different. This means that treatments for children should be tailored to their biology, so that they are more specific and effective.

We are interested in working out new ways to treat high grade gliomas in children; we know that these tumours are very diverse so a single drug or treatment may not work for everyone.

Understanding what makes these tumours different to each other, and different to the surrounding healthy cells in the brain, allows us a target the tumour cells specifically.

Targeting the tumour specifically in this way means that we can potentially reduce the dosage, limit side effects, and increasing the effectiveness of a treatment.

I will be looking at novel treatments, but also at using repurposed therapeutics, drugs that are already being used in the clinic to treat other diseases. Repurposed drugs are interesting to us because we already know that they are able to enter the brain and there is already data about safety and efficacy in patients.

We are very interested in combining different drugs to find the most effective treatments, using multiple drugs together has the potential both to increase the effect a single drug may have, allowing lower doses of each can be given, but also to reduce side effects.

My work will focus on the biology of paediatric high grade gliomas and how this might be targeted with different combinations of novel and repurposed therapeutics. Understanding which drugs work and why is the first step to getting new therapies into the clinic to treat patients with high grade gliomas.

 

What type of experiments do you do in the lab and what type of equipment do you work with?

I usually start each day in tissue culture, where we grow brain tumour cells to use in experiments. These cells are originally from patient biopsy which have been taken from a tumour and grown in plastic flasks in the lab.  The cells grow by attaching to the plastic surface inside the flask and are grown in nutrient media which contains all the nutrients and growth factors that the cells need to grow and divide. The cells are grown in an incubator, which mimics the conditions in the body, with stable temperature and carbon dioxide levels.

The cells need to be handling in specialised tissue culture hood which uses a circular flow of air to prevent any bacteria or other contaminants from coming into contact with the cells or media. Cells must be handled inside this hood at all times to protect both the scientist and the samples. We work in a specialised tissue culture room which is separate to the main lab and which we keep clean to minimise any potential contamination.

One of my regular experiments is make spheroids from the cells. I de-attach the cells from the flasks they have grown in and load them into plates which have a rounded surface and are treated with a coating to prevent the cells attaching. This forces the cells to round up and form a 3D ball of cells, rather than attaching to a plastic surface and growing in 2D.  Cells which grow attached to a flat surface behave differently to cells which are able to form spheres, these spheroids are biologically much more like tumours than cells which are grown on a flat surface. This means that the spheroids respond to drugs much more like a tumour, and I am therefore able to use the spheroids as a model to test new drugs, repurposed drugs and combinations of these drugs.

Once the spheroids are formed and treated with drugs I use fluorescent stains to measure which cells in the spheroid are alive or dead. The stained spheroids are imaged on a specialised fluorescent microscope which takes an image of each fluorescent stain and creates a combined image. Using this tumour spheroid model and the fluorescent stains, I am able to identify which drugs are killing the cells in the spheroid.

Once I have identified which drugs are killing the cells in the spheroids, the next step is to work out why they are dying.

The first stage of this is to measure the level of proteins in these cells, comparing the levels of proteins of interest in the drug treated spheroids with levels of the proteins in spheroids which have not been treated with drugs.

Understanding which proteins increase, or decrease, in the presence of a drug when compared to the normal level in untreated cells allows us to work out which pathways are being activated in these cells as a direct result of exposure to the drugs. Understanding the levels of proteins which are changed in drug treated cells, means we can track back to which pathways are being changed due to the presence of the drug or combination of drugs.

These types of experiment are a large part of the work that I do – by understanding which drugs kill tumour cells, but have no effect on healthy cells, gives us an understanding of how to approach new, or more effective treatments for paediatric high grade glioma.

I am particularly interested in drugs which alter a pathway called Autophagy (or ‘self-eating’), a pathway that all cells use to recycle old, or damaged cell components. Autophagy is relied on during times where nutrients may be restricted and as a means to remove damaged components which could be toxic if left.

Cancer cells, in particular, paediatric high grade glioma rely very heavily on this pathway to provide the nutrients they need and allow the cells to grow in stressful situations which would be toxic to healthy cells. This allows cancer cells to grow in hostile environments which may be low in essential nutrients. Blocking or otherwise altering this survival pathway in cancer cells has the potential to make them respond better to treatments.

 

What do you hope to achieve?

I am looking forward to learning much more about the field of brain tumour research as a whole, particularly the area of paediatric high grade glioma and I look forward to working with other scientist across different Universities as part of my studies. My project is currently at a very early stage, which means that I have some flexibility to explore and try out new things. As my project progresses and takes shape I am a looking forward to researching in depth novel and repurposed therapeutics.

I hope that my research will contribute in a small way to our understanding of the biology of high grade gliomas in children, and towards finding new therapies for use in the clinic. Paediatric high grade glioma is a really exciting field to be a part of, there is lots of research going on and I am very grateful to the Ollie Young Foundation for supporting my research.

 

How was it meeting the team?

I remember being really nervous before I went in to meet my supervisors, I was sat in the near-by coffee shop before the meeting getting worried; but actually there was no need. The team were really friendly and we talked about the project and our ideas for the work I would be doing at Portsmouth during my research project.

I left the meeting feeling really positive, the group at Portsmouth are really fantastic, they are all very knowledgeable and are keen to pass that on to train new research scientists in brain tumour research. Everyone is really friendly and welcoming and I am very pleased to be working alongside them.

 

What do you do in your spare time?

I like walking, cycling, running and swimming; and when I can I enjoy scuba diving.

I like to run mid- long distances, I really enjoy the training towards an event, as well as race day – running is a good way of keeping fit for me, but also means I am able to fundraise for charities which are important to me.

Currently I am training for the Great South Run which I am very pleased to run on behalf of the Ollie Young Foundation.