Models to study neuroblastoma in the laboratory

Finding suitable research models to study disease is a big challenge for researchers around the world. In cancer research, it is essential to work with models that can recapitulate tumour characteristics as much as possible. This is important to test chemotherapeutic drugs, understand tumour behaviour and have higher chances of translating the finds from the laboratory to clinical practice.  

Multiple factors influence tumour behaviour and disease progression. The most important is the tumour microenvironment, which comprises different cells and molecules that surround the tumour and the extracellular matrix, a network of molecules that provides support to the cells in the body.  

Most cell studies in a laboratory are based on 2D cell culture models in which the cells grow in a monolayer. Although this approach has a low cost and it is easy to use, it lacks the complexity observed in the clinical scenario. It is true that no model can recapitulate all the complexity found in the body. However, scientists were able to develop interesting approaches to study different tumour characteristics with relatively good approximation1.  

Specifically for neuroblastoma, the most common solid tumour that affects children, scientists developed 3D models in which neuroblastoma cells grow interacting with the surrounding environment and with each other in a vial. Examples of 3D models include cells grown in hydrogels or scaffolds and multicellular tumour spheroids (see image below). Spheroids are formed through the self-adhesion of tumour cells growing in the form of very small balls. They can be maintained in the laboratory on their own or supported by scaffold-based platforms (jelly-like or porous materials). Scaffolds essentially support the cell resembling the extracellular matrix and surrounding tissue in the body. 

In the Cancer Bioengineering Research Group, we work with neuroblastoma models such as organoids, a more complex type of spheroid, to understand neuroblastoma migration and invasion2. Moreover, we recently shared with the research community a protocol at jove.com describing the development of a 3D neuroblastoma model using collagen-based scaffolds3.  

Time-lapse video of neuroblastoma organoids’ growth. Accompanying experimental data published in Gavin et al., Cancers 2021. Source: the Cancer Bioengineering Research Group 

These models have the potential to advance drug tests performed in the laboratory providing better clinical translation, ultimately contributing to improving the quality of life and survival of children diagnosed with neuroblastoma.  

The work with 3D models at the Cancer Bioengineering Research Group is supported by the Irish Research Council, the Conor Foley Neuroblastoma Cancer Research Foundation, Neuroblastoma UK and National Children’s Research Centre. 

Written by Luiza Erthal

References 

1. Nolan, J. C. et al. Preclinical models for neuroblastoma: Advances and challenges. Cancer Lett. 474, 53–62 (2020). 

2. Gavin, C. et al. Neuroblastoma Invasion Strategies Are Regulated by the Extracellular Matrix. Cancers 13, 736 (2021). 

3. Gallagher, C., Murphy, C., O’Brien, F. J. & Piskareva, O. Three-dimensional In Vitro Biomimetic Model of Neuroblastoma using Collagen-based Scaffolds. J. Vis. Exp. 62627 (2021) doi:10.3791/62627. 

A 30km Dublin Mountain Way in A Day

And the story began with a meeting of fantastic 7 at the very beginning of Dublin Mountains Way in Tallaght at 6.30 am on September 25th. The spirit, cheer, backpacks with essentials and branded tops were on, Strava was launched and we swiftly headed off.

It was quiet, dark and cheering. No one was on the streets, a few cars passed by. We took towards Bohernabreena reservoir through the sleepy estates of Tallaght, sensing the sunset. Clouds were low and the highest peaks in the Dublin Mountains including Seefingan, Corrig and the highest, Kippure were in the mist. Nevertheless, we were full of energy and hopes to see it later.

Cheat chats and jokes were here and there, we walked in small dynamic groups recalling our pre-covid life and stories that happened during the lockdown. A mix of newbies and maturating research students. We met some in person for the first time since the COVID restrictions admitting that our visual senses are extremely important to memorise a person and recognise him/her on the next occasion. We were enjoying this face-to-face communication and our team re-connection.

The first 8 km flew in a flash. We stopped for our breakfast in Dublin Mountains. The grass was wet, the sky was blue. Mountains started to draw their shape through the clouds. Yoghurts, fruits, bars immediately disappeared in our stomachs. Everyone was happy to lighten their backpack. Every little helps!

A few plasters were glued, and we continued on at a very good pace. The sky was changing with sunny spells. We travelled around Spinkeen and Killakee at their base doing up and downhills and verifying our route with the hiking app. At the 20 km mark, we stopped for lunch. Sandwiches, grapes, mandarines and sweets were shared and eaten and then polished with chocolates from the recent Nadiya’s home trip. Jellies left untouched.

At 25 km, our blisters reminded us of being humans. Our pace slowed down and we started a very mild ascent to Tibradden Mountain leaving the Pine Forest or Tibradden Wood behind. We climbed further to Fairy Castle, the highest point on the Dublin Mountains Way (537m). Throughout the entire way, Dublin showed its best views of the Phoenix Park and the Pope Cross, house roofs, Aviva Stadium, two Chimneys, Dublin Port… The scenery was fascinating and breathtaking. We saw Howth and Dun Laoghaire, Sugar Loaf… We met groups of Germans, French, Irish and many others.

At Three Rocks Mountain/Fairy Castle, we started our descent and entered Tiknock forest. This part was steep. We crossed the Gap Mountain Bike Adventure Park to reach Glencullen. Got lost at the end but just for a sec and reached the Glencullen junction at 2.30pm. It took us 8 hours with walks and stops from start to finish to complete the 30 km challenge in a day. We got tired but felt happy and satisfied.

We aimed to raise awareness of childhood cancer in general and neuroblastoma in particular as well as honour children with cancer, their parents, siblings, friends and careers, doctors and nurses, volunteers in the hospitals and researchers working to find cancer weaknesses and develop new treatments that are friendly to patients and target cancer aggressiveness.

We will count our tally in the coming days and transfer it to three wonderful charities that support childhood cancer research.

We thank everyone who supported this challenge!

Go raibh maith agat!

Dublin Mountain Way in A Day, September 25th 2021

Here are our plans. This year we have upped the challenge, taking on the Dublin Mountain’s Way in a Day ⛰ We will hike through the Dublin Mountains from Tallaght to Glencullen, and maybe even all the way to Shankill on September 25th! Our challenge is not only to do #DMW in a Day & support three wonderful charities CMRF Crumlin/National Children’s Research Centre, Neuroblastoma UK and the Conor Foley Neuroblastoma Cancer Research Foundation but also beat our past fundraising records! If we raise 2K+, we’ll do 30km in a day. If 3K+ then 42km! Can u challenge us?  All funds raised will go to the 3 selected charities. Every donation big or small is hugely appreciated!

Please support us by donating to our Gofundme

https://gofund.me/ec59f131

Childhood Cancer Awareness Month 2021

Every 100th cancer patient is a child. Cancer is the 2nd most common cause of death among children after accidents. 

Childhood cancer is an umbrella term for many other types of this disease. Every September, many charities, researchers and parents of children with cancer work hard to raise awareness of this cancer. You may learn more about kids with cancer, their loving families, the doctors and caregivers who looking after them and treating them, the young survivors of cancer and those kids and teens who lost their battle, and the scientists who working hard to find a way to stop childhood cancer.

This year our research team will hike Dublin Mountain Way in One Day on the 25th of September 2021 whatever the weather in honour of Childhood Cancer Awareness Month. For every one euro donated to research only 1 cent of this goes to ALL childhood health conditions including cancer. Therefore, the donations we receive will be split equally among some wonderful children’s charities. These charities include the Conor Foley Neuroblastoma Research Foundation (CFNRF), Neuroblastoma UK (NBUK), Children’s Research & Medical Foundation (CRMF) Crumlin.

If you would like to get involved in this amazing challenge and help us raise vital funds for childhood cancers, you can contribute to our fundraising page:

In the Spotlight at EACR Virtual Congress

In early April, shortly after attending the Irish Association for Cancer Research (IACR) virtual conference, I was delighted to find out I had been selected for a “Poster in the Spotlight” presentation at the European congress in June. I was invited to give a 10-minute presentation about my research on finding new immuno markers for drug-resistant neuroblastoma. This would be my first time presenting at a conference outside of Ireland (though I remained in my home in Dublin and not in the hoped destination of Turino) and so it was a very exciting experience!

On the morning of my talk, I logged onto zoom to meet the EACR session organisers and the other speakers in my session – a mixture of PhD students and Post-doc researchers from Germany, Spain, Italy and the UK. Each of our talks was then broadcasted to the virtual congress platform where cancer researchers around Europe could tune in.

After the presentations, we joined a “Meet the Speakers” session where we could chat with those who had tuned in, answer questions and open up some research discussions. I was asked one question from a researcher doing similar work to me on how I investigate the relationships between certain genes in cancer, and I was able to refer her onto the software I use – I hope this aids her research, as that’s the real aim of attending these conferences!

Following this, it was time to relax and watch some of the other interesting talks taking place in the congress. The EACR were running a photo competition to show where you were watching from in order to win a place at the 2022 congress in Seville. So, I took this opportunity to take my laptop out to the back garden and watch the congress in the sun, getting a selfie for twitter with #NotQuiteTorinoEACR2021. Fingers crossed by this time next year travelling for international conferences will be a reality for researchers once again!

Catherine Murphy, Neuroblastoma UK funded PhD student

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

International Childhood Cancer Awareness Day 2021

Every year, we celebrate Childhood Cancer Awareness Day Internationally. Pre-COVID times, it was straightforward to do a coffee morning, a bake sale, or as we did a Hot Chocholate morning.

To be honest with you, I had almost no presence of childhood cancer in my life until I joined Prof Stallings lab in 2011. When I said ‘almost’, I meant during my adulthood.

My Dad had a younger brother, both started their families at the same time. Our families used to spend a good time together, holidays, birthdays and weekends together. Both I and my cousin Igor were born almost within a year. I remember our play days together but at the level of feelings and stories told by my parents. In the pic, we were held by my nanna.

I do not have a pic where we were 3- or 4-years old. No pic was taken after Igor was diagnosed with blood cancer. He was 1-1.5-year-old. He travelled 800km to the best paediatric oncologists to receive the most progressive treatment back then. It extended his time with the family by four years. But he lost his battle…

How much can remember a 1-, 2-, 4- years old? My last memory – Igor was sleeping in a neat coffin. Adults were muttering. I remember, a tear slowly rolled down from Igor’s closed eyes. I naively asked my nanny why did the dead cousin cry??? He did not want to leave us, – said my nanny softly. 

Back then parents were not much informed on the disease, treatments, odds and alternative options. Igor was suffering, no pain relief options were available… No palliative care… Remember my nanny’s words: “These doctors had no hearts. All this child needed was a sense of peace, quiet time with his parents away from the hospital wards”. 

Many things have changed since then. Eight out of 10 children with blood cancer are responding to treatment well, they reach adulthood, may even have kids of their own. However, there are some types of childhood cancers that do not respond well and can return to being more aggressive. Cancer steals the child’s future. One of the thieves is neuroblastoma, a solid tumour of undeveloped nerves.

Childhood cancer research is essential to return happy days to kids and their families. Many childhood cancer research charities do their best to secure funds and support researchers like me. It is vital to have a continuous investment in research that helps to understand the weakness of childhood cancer and develop new drugs designed exclusively for kids.

Today I want to thank 3 charities for their hard work: Children’s Health Foundation Crumlin, Neuroblastoma UK and the Conor Foley Neuroblastoma Cancer Research Foundation. And ask you, my readers, to donate to a childhood cancer charity of your choice.

10 Laps 10km: We raised a grand total of €1419

Last month we set ourselves the “10 Laps 10km” challenge for Childhood Cancer Awareness.

Now we have closed the GoFundMe and counted the charity buckets. We are delighted to announce we raised a grand total of €1419! We are over the moon with this sum, as 2020 required a very different kind of fundraiser than previous years. 

Our three chosen charities: Children’s Health Foundation Crumlin (formerly CMRF), the Conor Foley Neuroblastoma Cancer Research Foundation, and Neuroblastoma UK, will each receive just over €470. 

We’d like to say big thank you to everyone who donated. It will make a huge difference for these charities, this year especially, paving the way to better treatment options for children with cancer in the future.

10K Vhi Women’s Mini-Marathon – #POWEROF10

The new norm, new challenges, new excitement and new achievements! We all proud to say that we completed 10K Vhi Womens Mini marathon socially distanced. Our paces were so different that distancing came absolutely natural. We ran it individually but were a team mentally. Even the capricious Irish weather was our ally. The Sun was bright. The air was fresh and crispy.

This was an individual challenge #POWEROF10: just you and the trail. 10 laps around St Stephen’s Green park were to make the target 10K in aid of Childhood Cancer Awareness Month. The celebration of life, therapeutical advancements, the strength of little patients battling their cancer and their families, doctors and carers who are supporting them in their journey. The emphasis on the gaps in available treatments and diagnosis and the importance of research that needs funding.

Personally, my 10K were split into two parts. The first 4K were full of arguments with my body. Why didn’t I like to do laps? Could I complete 10K? Was I fit to do it? Keep going! No walking – better slow jogging. Did one lap make 1K? Should I do a longer lap instead? And so on and so forth. Then, the second part kicked in. My body stopped arguing and began to enjoy it. I noticed beautiful Autumn colours on the trees, people walking around with a cup of coffee or chatting away, saw my team members overtaking me, and our volunteers counting our laps. People on the street and in the park were cheering us up. What a wonderful and fulfilling day!

As Catherine says: “The 10 Laps 10km challenge was tough! Like many people, I took up running casually during the lockdown, however, I never did more than a couple of kilometres at once, so I was absolutely not prepared for running 10. But the cheers from our socially distanced spectators and all the online support we received meant I got through it. Also knowing what a positive impact this challenge could have for the future of childhood cancer treatment provided plenty of motivation to finish the race 💛🎗”

10K by 6 team members socially distanced. #POWEROF10. Go Gold! Let’s reach 1.5k in donations!

Our Go Fund Me page is still open until this Sunday (October 11th midnight) if you wish to support us.

One Day of the Life as a Researcher: PhD student

A lot has changed for me since I began my research journey in RCSI, as I transitioned from being an undergraduate placement student to a PhD candidate, however the biggest change has been adjusting to doing a lab-based PhD during a pandemic! 

We wear red coats when looking after our rather large family of neuroblastoma cells, which happily grow inside the 37°C incubator. White coats are for most other lab work, such as analysing proteins by gel electrophoresis and Western blotting. And of course, the newest lab accessory – the facemask.

These days my work hours are shared between the labs in RCSI and my family home. While my bench space and office space used to be separated by just a few steps, there is now a 30+ minute bus journey between them. It has certainly put my planning skills to the test as now when I walk into the lab I need to be sure of what I am planning to do, and that I can complete the task in my pre-booked lab time slot. 

I appreciate my time in the labs much more now that I spend so much time at home. Whether I am culturing neuroblastoma cells, analyzing proteins or genes by Western blots or PCRs, I enjoy immersing myself in the work knowing that my time on the bench is limited. 

The main perk is that now when I am doing computer work – analyzing results, writing reviews, preparing presentations, using online software – I can do it from the comfort of my box-room-office, often very cosy in a blanket as I do it. While my work-from-home desk space is slightly more spacious than my desk in the now-closed “Write-up Room 2”, I do miss the chats and laughs that come with working in a shared office.

One thing’s for sure though, my two dogs very much enjoy the days that I work from home!

Catherine Murphy, Neuroblastoma UK funded PhD student