This was our 2nd time attending the OLCHC Research & Audit Day on May 25th, 2018. The conference provides a great forum for paediatric clinicians to share and update knowledge across different specialties through talks and poster presentations. It is insightful for basic biomedical researchers like us to see other perspectives.
I was delighted to know that two our studies were shortlisted. It is a rewarding feeling to see your Dream Team doing very well. One was the project of the Erasmus+ student Hanne Pappaert and the other was the project of NCRC funded Postdoc John Nolan. Hanne explored our 3D tissue-engineered model of neuroblastoma using collagen-based scaffolds with distinct mechanical properties. These new scaffolds were designed and manufactured by our collaborator Dr Cian O’Leary from Pharmacy Department and Tissue Engineering and Research Group (TERG) headed by Prof Fergal O’Brien. Hanne grew 5 neuroblastoma cell lines on the 3 scaffolds: hard like a rock, soft and fluffy like a cotton wool and a jelly-like. All cells liked the jelly-like environment. This environment is similar to bone marrow – the most common site of neuroblastoma metastasis. We were excited to see the difference as it means we are one step closer to reconstruct this type of tumour spread.
John has expanded our exploration of our 3D neuroblastoma model by examining the content of exosomes – little parcels sent by cancer cells in 3D and as tumours grown in mice. We were thrilled to see a high similarity in the exosomal content. This finding additionally proved the great applicability of our 3D model as a tool to study neuroblastoma.
Though the official announcement is scheduled for the first week of June, the groundwork is on. Lots of reading and planning for the trip to Johns Hopkins later this year. One of the first is the book by Rebecca Skloot ‘The Immortal Life Of Henrietta Lacks”. The famous HeLa cells were generated by researchers at JH. The story is a fascinating journey for biomedical scientists and a tragedy for the Lacks family.
Here is the perfect example of the teamwork troubleshooting protein extractions. My Dream Team 2018 in action. The current information and communication technologies allow to stay connected and respond quickly.
Five minutes later in the lab: troubleshooting is the exchange of experiences!
Our body has 3 dimensions: height, width and depth. Every single part of our body grows in the same 3 dimensions. This is true for cancer cells. Researchers use different ways to study cancer cells behaviour, how they grow and spread. We grow cells in the flasks, where they change their structure and shape and become flat losing one dimension. This is a very popular approach. We also grow cells in mice, where cells keep their 3D shape and mimic their behaviour to one observed in humans.
It is well known that we need to give a different amount of drug to kill cancer cells grown in flasks and in mice. This, in turn, delays the development of new drugs. Why does it happen this way? So, the drug works only on one side of the cell when they grow on the flat surface. In contrast, in mice, drug surrounds the cancer cell habitat and attacks cells at the edge first and then getting to those at the core. So we need more drug to kill cancer cells in mice.
We decided to design a new way to grow cancer cells that recreate their growth in 3 dimensions as in the human or mice body. We used special cotton wool like sponges as a new home for cancer cells and populated them with cancer cells. At the next step, we gave cells the drug at the different amount and checked what happened.
To understand cell fitness we stained them with red and blue dyes. On the left bottom side of the image, we see an equal amount of red and blue dyes telling us that cells were healthy and fit. Cells did not get any drug. When we gave a little amount of the drug but enough to kill cells in the flask, the balance of red and blue dyes was the same telling us that nothing really happened (the image in the middle). Cells were feeling well and healthy. The right bottom image has only blue dye. In this case, cells were given the amount of drug enough to destroy cancer cells in mice or humans. The lack of red dye tells us that this time the drug worked and killed the cancer cells.
We found that the drug killed cells on sponges only at doses enough to do the same in mice.
So, we concluded the new tactic to grow cancer cells in 3D on cotton-like sponges can bridge the gap between traditional way and animal models. This new strategy to grow cells on sponges should help to understand cancer cell behaviour better and accelerate the discovery and development of new effective drugs for neuroblastoma and other cancers. This, in turn, will make the outlook for little patients better and improve their quality of life.
This work has been published in Acta Biomaterialia and presented recently at the Oral Posters Session at the 54th Irish Association for Cancer Research Conference 2018.
The fact of being shortlisted is very encouraging. It means that my research proposal and the career achievements fit the merit of this award. No doubt it was fantastic experience overall, not often the shortlisted candidates have an opportunity to speak for themselves.
How is it feeling after the interview?… Well, I do not have a firm answer… It is a big difference when you explain yourself in written and spoken forms… No chance to edit your real-time talk… How many times have that 30 minutes played back and re-run in my head? I lost the count… Did I bring the point across? Did I do things in right time and at a right pace? Should I have structured the answer differently? Each re-run brings new ways to answer the same questions, indeed, in a better and more concise way… Having the mind that is constantly analysing the situation is not helpful.
Think, the competition was very tough, and only 1 in 10 made to the 2 days interview for the Fulbright Award (maybe the ratio even higher). Twelve candidates were interviewed yesterday and the same numbers are to be today. What are the chances to get to the final? I have to wait until March… and meanwhile, keep applying for grants and doing something meaningful.
My next stop is at the Irish Cancer Society this Thursday to film a short video about my research and neuroblastoma challenges. The video should be available for the International Childhood Cancer Research Day on February 15th.
The first month of the new year and the first challenge. Monday is the big day for me. In the essence, my grant application was shortlisted for the interview where I have to face the challenge and prove that I worth it.
The proposed application seeks to go to the US lab and gain an expertise in an interdisciplinary methodology to monitor and capture the dynamic of cancer spread (metastasis) in real time. This experimental approach would accelerate our understanding of neuroblastoma metastasis which is one of the reasons for failure in the treatment of neuroblastoma. If we know how neuroblastoma cells spread, then we can find the weaknesses in the process and create new drugs or use existing to target it.
I feel that sharing my worries with you makes me stronger. I am looking forward to this challenge with my head up and hope to feel your support at this crucial moment.
When I look back on my journey in 2017, there were many junctions, traffic lights and stops as well as ups and downs. Junctions were to make decisions, while traffic lights and stops – to be patient. Ups and downs were my feelings of satisfaction. The good mix of both kept me to stay human. It is not the number of grants received that matters it is who around you. I have met genuinely curiosity-driven students who made this journey fascinating and very special.
My most memorable Ups were the successful examination and graduation of my PhD student John Nolan, organisation and chairing the IACR Meeting session: Challenges in Childhood Cancers, the Mad Hatter’s Tea Party and the Gala Dinner with the CFNCRF, the launch of my very own research team thanks to the funding by the NCRC and the Neuroblastoma UK, the successful completion of two final year undergraduate and two MSc projects, and welcoming the new PhD student Tom Frawley.
My team is growing and I am looking forward to 2018!
The researcher’s path includes days when you feel so low because your grant or paper was rejected or even both within a very short time frame. It happened to me a couple of weeks ago. At this point, I felt helpless sarcastic and non-motivative reading reviewer’s comments. One reviewer mixed up neuroblastoma with a brain tumour, so their comments were not relevant. Another just found no time to read through, the sentence was very short – ‘not a priority or interest‘. One more went to their area of expertise asking to fulfil it rather than comment on the actual focus of the study. Such comments are so common that any submission of results or a proposal could be considered as a draw. It has been neither my first time not the last. More to come.
On November 20th, the Irish neuroblastoma researchers have met for the first time to set up a collaborative research hub. The aim is to consolidate their expertise and skills in order to crack the neuroblastoma code together.
They all have different science background spanning from molecular and cellular biologists, immunologists, tissue-engineering, bioinformatics, mathematical modelling and clinicians representing RCSI, UCD, TCD, OLCHC and NCRC. During this meeting, researchers talked about their challenges and progress finding out that we are complementing each other projects. Clinicians from different OLCHC departments exposed basic researchers to realities of the disease. None would find this information in academic papers: it is what you see in the clinic and how it works in practice.
Big thank you to Dr Cormac Owens for the invitation and linking us together and Prof Jacinta Kelly for mapping the support available from the NCRC and CMRF.
Our next meeting will be held in RCSI in January 2018.
Happy Birthday the Irish Neuroblastoma Research Consortium!
As a part of Science Week, our Department hosted Lab Safari for the secondary school students aiming to give a sense of what scientists do in the lab. The focus of my research is the identification of novel biomarkers for neuroblastoma response to chemotherapy. So, we decided to explain the concept of biomarkers and the importance of discovering novel biomarkers for neuroblastoma. Between me and John we covered biomarker’s basics and carried out the detection of MYCN in neuroblastoma cell lines. Here, I am summing up our activities in the form of Qs & As:
1. What are biomarkers?
Biomarkers are signposts of the body condition. Biomarkers – bio is for biological and markers – for molecules indicating that something is going wrong or differently in the body. They can help doctors to read these changes and identify a condition or disease. There are biomarkers for heart disease, cancer, diabetes and many others.
2. What type of molecules can serve as biomarkers?
Various types of molecules can do the job as long as they can differentiate the normal and abnormal process in the body. It can be DNA, RNA, proteins and hormones. These molecules can be detected in the blood, urine, stool, tumour tissue or any other bodily tissues or fluids.
3. How can we discover novel biomarkers?
Research, research and again research. We have to learn more basic facts about DNA, RNA and proteins. We need to be able to link this knowledge with health conditions including cancer. We need to identify how these molecules speak about the unhealthy conditions. We need to explore whether biomarkers exist within the disease that can predict the response to treatment and outlook.
4. What biomarkers are known in neuroblastoma?
There are various biomarkers that are specific for neuroblastoma. Catecholamines in urine are chemicals produced by neuroblastoma cells. Chromogranin A – is a protein that can be detected in the blood. It is unspecific biomarker because it can be secreted by neuroblastomas and some other tumours (e.g. pancreas and prostate cancer). Various chromosomal aberrations can be biomarkers of neuroblastoma aggressiveness. One of the strongest predictors of rapid neuroblastoma progression is MYCN status.
5. What is MYCN?
The MYCN is a proto-oncogene protein and a member of the MYC family of transcription factors. MYCN was identified in 1983 and very soon became a routine biomarker for neuroblastoma aggressiveness. This protein is very important for the normal embryo development. The amplification of this gene leads to the excess of MYCN protein in cells prompting cells to grow and divide faster transforming normal cells into tumour cells.
6. How did we detect MYCN in neuroblastoma cell lines?
A day before the demonstration, we carried out some preparation steps. We selected several neuroblastoma cell lines with known MYCN status. Some cell lines had MYCN amplification, other didn’t. Among those that did have the amplification, the expression of MYCN protein was different giving us a good illustration of biomarker’s quantity. So, we run SDS-PAGE, a technique that helps to separate the cellular protein mix based on their size.
Then the separated proteins were transferred onto a membrane and exposed to a buffer containing an antibody for MYCN (primary antibody). This antibody recognises only MYCN. At the next step, we added another antibody that has a dual role – it can attach to the primary antibody and produce a signal that can be visualised. The next step would have been the visualisation itself and the one for Lab Safari. So, we were ready.
Once we finished the basics of biomarkers, we moved to the biomarker MYCN detection step. The buffer for visualisation was added and the membrane was imaged.
If you like this idea and want to have this demonstration for your science classes at school, please contact me and we would be happy to do it for you.