Sparkles of A Researcher Day

Once I mentioned the importance of the publication track record for a career in science. My team has been productive despite the COVID pandemic. Two review articles were published.

The first was a review written by Tom and published in Cancers focusing on the small extracellular vesicles produced by cancer cells that can transfer various growth signals to the tumour microenvironment aka neighbourhood and promote tumour expansion. The signal in focus was a protein called epidermal growth factor receptor (EGFR). It contributes to the healthy fitness of many different cells in the body. However, many cancer cells produce an excess of this protein giving them an advantage of growth over normal cells. Increased EGFR can be seen in breast, lung, glioblastoma and head and neck cancers.

Cancers 202012(11), 3200; https://doi.org/10.3390/cancers12113200

The second review has been published in Journal of Personalized Medicine on March, 16th. Originally, it was a small review project for Nadiya, a medicine student, last summer. However, it became a big one with all data systematically collected, analysed and condensed. The focus of this review was on Retinoic Acid (RA), widely known as Vitamin A and its role in neuroblastoma. RA plays a vital role in human development. The main feature of RA is to push neuroblastoma cells to become neuron-like cell stopping their aggressiveness and cancer fate. So, we wanted to know more about the ongoing research both in the labs and the clinic. We reviewed primary research articles reporting basic and translational findings as well as clinical trials. Hopefully, it would help other researchers to get a full picture of this topic and a structured resource of experimental models and drugs tested.

J. Pers. Med. 202111(3), 211; https://doi.org/10.3390/jpm11030211

Annual NCRC Symposia 2020

As the year comes to an end, you are looking back and seeing all achievements in a different light, a light of the COVID glaze. Lab research was at bay for a while, challenges to return and re-start experiments, no scientific meetings in the traditional format where you build your new collaborative net at coffee breaks. Despite all, the team has expanded and we welcomed Ellen and Erin in October.

The NCRC Winter Symposia is a lovely way to wrap the year putting together all hard work and look at the progress done so far. We have an exciting project that has two arms: a blue-sky science and a translational. Working together John and Tom were able to generate promising results on understanding how small membrane-bound vesicles or exosomes can send signals from neuroblastoma cells to cells responsible for new blood vessels formation. They developed a protocol to scale up the production of exosomes, isolate them and characterise. We have a dataset on what these exosomes carry on and now can test how they promote new blood vessels formation. Indeed, more left to do but knowing the direction makes this journey meaningful.

Hallmarks of Research

Research is a fascinating journey no doubt. Inquisitive minds try to solve burning puzzles. It takes time. Some puzzles are more complected than the others. One of the hallmarks is the conversion of the resolved puzzle into a scientific story to tell to your peers.

We write and publish these stories. The publishing is another caveat that often makes your story sharper and neater. However, while you are in the process you feel that the mission is impossible.

Delighted to see that one of the missions is completed – a great hallmark for John which coincided with his new research adventure starting in a few days. This is his first first author paper! It is not tautology! It is his first original research paper where he is the first author. This position is a success measure in a research career. His teamwork skills secured him another few original papers. Well done John! Well deserved!

This study is an excellent example of the many roles that small RNA molecules such as miR-124-3p can play in neuroblastoma pathogenesis. The ability of this miRNA to work together with standard chemo drugs can be exploited further in the development of new anticancer therapeutics targeting relapse and drug-resistant tumours.

Preclinical models for neuroblastoma: Advances and challenges

What a great start for 2020! Our long-lasting and productive collaboration with our colleagues from Tissue-Engineering Research Group Brough to live an important overview of the preclinical models for neuroblastoma. We particularly focused on the 3D in vitro models available.

During this exercise of searching and reading research papers, we found that researchers in neuroblastoma are looking for alternatives of traditional 2D culture. It is may be slow at the moment but the interest is there.

3D neuroblastoma models worked well in both validating known chemotherapies and screening new. The concepts and materials that were initially developed for bone or tissue regeneration can be used to a miniature model of neuroblastoma.

3D tissue-engineered models can accelerate drug discovery and development, reducing the use of animals in preclinical studies.

Full version is available at https://www.sciencedirect.com/science/article/pii/S0304383520300239?via%3Dihub

Scientific part of my journey

Reading my posts, it looks like I am more enjoying the cultural part and almost forgot the main reason I crossed the Atlantic with the Fulbright wings.

The first month in the lab was more a warming up. Where is my desk? Where is the cell culture rooms? How do they run it? How different is it? So, many microscopes – am I capable of imaging? And so on and so forth…

My typical day starts at 8-8.30 am and finishes once all is done. It may be 6pm or 10pm. Once the experiment is set up, I have to monitor cells every 24 hours for 5-7 days with no weekends or days off. The monitoring includes imaging. Lots of imaging. Every condition has 20-30 single cells to follow up. Each cell gets its own GPS tag manually to be able to image exactly the same cell as it grows and becomes a group of hundreds by multiplication. For example, I am running 8 different cell lines in 3 experimental conditions. So, 20-30 cells per all 24 combinations give us 480-720 individual cells to follow up. The imaging takes ~5 hours every day. After 5 days, I will have 2400 – 3600 pics of cells to analyse. It will be fun! I may need lots of Guinness to fly through that numbers.

Tagging cells. The left arrow points to a group of neuroblastoma cells. The arrow in the middle point to the same cells, but this image allows you to see the actual number of the cells. This group has 8 cells. The right arrow points to individual GPS tags for each cell

At the next step, I will select some of the conditions for video recording to trace cell fate from a single neuroblastoma cell to a metastatic niche consisting of hundreds of them. This video will show me how it all happens minute after minute.

Is not it exciting? I am thrilled!

 

8th OLCHC RESEARCH & AUDIT CONFERENCE

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.

 

How current IT advances help in research?

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!

 

A new 3D strategy to study neuroblastoma

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.

This study was supported by Neuroblastoma UK and National Children’s Research Centre.

You can find more at

A physiologically relevant 3D collagen-based scaffold–neuroblastoma cell system exhibits chemosensitivity similar to orthotopic xenograft models.

IACR Meeting 2018 Programme

Irish Neuroblastoma Research Collaboration

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!

 

 

 

 

 

 

What lessons have been learnt?

Today is the final day of the Third International Cancer lmmunotherapy Conference. The meeting was run at the Rheingoldhalle Congress Center in Mainz/Frankfurt, Germany from September 6-9, 2017. More than 500 people attended this meeting.

The focus of the scientific program was on “Translating Science into Survival”. Talks covered the challenging areas in cancer immunology and immunotherapy. The full list of topics can be found in the meeting program.

At the moment cancer immunology and immunotherapy is a hot topic in the next generation of anti-cancer therapies. Lots of attention is given to checkpoint immunodrugs as it was proven by the prevalence of talks on this subject in the program. Indeed, this drug has great potential, but at the same time, it is not universal. About 50% of patients do not benefit from it.

What lessons have been learned from the talks:

  • Checkpoint immunotherapies are the main stream
  • Not all cancer patients would respond to immunodrug
  • Genetic landscape of a tumour and/or the patient may contribute to this, thus making beneficial to check genetics for this type of treatment
  • Immunodrugs work better in combination with conventional therapies such as chemotherapy.
  • The immune system can be tuned by a drug, but it will switch on compensatory mechanisms to balance the intervention.
  • Lots have to be studied further