A new, three-dimensional approach to cancer research

Appeared in today’s Irish Times. Lovely crafted by Dr. Vanesa Martinez

Although the discovery could be applicable in principle to any a solid tumour, Dr Piskareva’s target is neuroblastoma, a relatively common child cancer which affects a specific type of nerve cells in unborn children. “It’s quite aggressive and unfortunately there are many children who have metastasis when they are diagnosed, and this is the most challenging group to treat.”

Irish Times, 31 May 2018

https://www.irishtimes.com/news/science/a-new-three-dimensional-approach-to-cancer-research-1.3505347

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.

 

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

The First Big Challenge in New Year

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.

Anyone for a challenge?

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.

 

 

 

Neuroblastoma Research Dream Team 2017

It is fantastic to see so knowledgeable and enthusiastic young researchers in my research group. This year, the team is multinational with the Irish students mixing with Belgian and Malaysian. All together they are cracking the code of neuroblastoma microenvironment and tumour cells communication through understanding main differences between conventional cancer cell models and tumours.

The big research plan of the entire team consists of more smaller and focused projects to be completed within 10-12 weeks. All projects are unrestricted, they are driven by the intellectual curiosity of these students. This way is full of ups and downs, frustrations and encouragements when techniques do not work or reagents do not come in as expected. Some cancer concepts can also work differently in the given settings. Simple questions are bringing more challenges than expected.  But at the end of the road is the best reward – contribution to the conceptual advancement of neuroblastoma microenvironment.

 

 

The Neuroblastoma Research Dream Team 2017: Dr. John Nolan, NCRC funded researcher, RCSI, Joe O’Brien, TCD MSc student, Ciara Gallagher, DIT undergraduate student, Jessica Tate, RCSI Medical student, Larissa Deneweth, Erasmus student, Ghent, Ying Jie Tan, TCD MSc student.

Feeling good, excited and accomplished

This week can be rated for sure as feeling good, excited and accomplished. A UK based charity – Neuroblastoma UK has awarded a small grant to characterise a pre-clinical model of neuroblastoma which is a collaborative project between our lab and Tissue Engineering Research Group at RCSI. This project will study features of neuroblastoma cells growing on collagen-based scaffolds. The NBUK grant will contribute to one of the most expensive parts of the study – characterisation of cell secreting proteins using antibody-based profiling platforms.

Another research was accomplished yesterday –  John Nolan had his Voice Viva examination and successfully defended his PhD Thesis. This 3 year PhD project was funded by the National Children’s Research Centre. As his supervisor, I am delighted for him and wish him best of luck in his research career.

 

 

Cells having a handshake in 3D

Neuroblastoma cells growing on scaffolds.

Continue research into 3D neuroblastoma models, we imaged cells growing on collagen based scaffolds using confocal microscopy. This technique is very popular in cell biology providing depth in cell imaging.

Here you can see cells growing on scaffolds: white dots – cells, irregular fibers – collagen containing scaffold.

 

 

The results are fascinating! Cell nucleus is in blue (DAPI), cell actin is in red (phalloidin). You will be able also to see how two cells ‘having a handshake’. It is happening just in the middle.

Towards 3D neuroblastoma cell models

It seems I have got a conference season. Three conferences within 2 months – no complains though. This time I went to the Matrix Biology Ireland Meeting in Galway. It was fantastic mix of topics and speakers ranging from new approaches in bone and heart repair to new matrixes in reconstruction of body tissues and diseases in the lab to minimise use of animals in pre-clinical studies.

My talk was focused on neuroblastoma microenvironment and cell-to-cell communication through exosomes. I wrote about it in October post. I talked about things that did work and did not as well as new directions. One of the new directions is reconstructing neuroblastoma by growing neuroblastoma cells on collagen based scaffolds in 3D. Collagen constitutes most of our tissues to keep it shape and strength. These scaffolds are sponge-like matrixes built from collagen and other components. Of course cells grow differently on these matrixes. They have a different shape and growing properties in 3D. Neuroblatoma cells look like water drops on the cotton wool-like collagen scaffolds. In contrast, when they grow on plastic in 2D, they are flat. Studies show that cells in 3D respond to cytotoxic stress in a similar pattern as if being within a body (details in recent reviews 1-4).  It would be a great breakthrough once these models are optimised for neuroblstoma research  field. It will help to test all new and known drugs in the environment close to clinical settings. It could be a step forward to personilised therapies for children with neuroblastoma by isolating cancer cells, growing them in 3D and testing how they respond to all therapies available. It will facilitate more efficient design of treatment for relapsed or poorly responding tumours, sparing patients unnecessary rounds of chemotherapy and ultimately increasing survival.

 

Neuroblatoma cells look like water drops on the cotton wool like collagen scaffolds. In contrast, when they grow on plastic in 2D, they are flat. Arrows point towards cells.
This is microscopic images of neuroblatoma cells growing on the collagen scaffolds and plastic. Arrows point towards cells.

 

I’ve always felt that a selection of abstracts for an oral presentation is biased. The overall background and views of conference organisers would affect works selected for an oral presentation.  The same abstract was not selected for an oral presentation by one committee, but was supported by the other.  Never give up!

Readings:

  1. Schweiger PJ, Jensen KB.Modeling human disease using organotypic cultures. Curr Opin Cell Biol. 2016 43:22-29.
  2. Salamanna F, Contartese D, Maglio M, Fini M. A systematic review on in vitro 3D bone metastases models: A new horizon to recapitulate the native clinical scenario? Oncotarget. 2016 7(28):44803-44820.
  3. Picollet-D’hahan N, Dolega ME, Liguori L, Marquette C, Le Gac S, Gidrol X, Martin DK. A 3D Toolbox to Enhance Physiological Relevance of Human Tissue Models. Trends Biotechnol. 2016 34(9):757-69.
  4. Nyga A, Neves J, Stamati K, Loizidou M, Emberton M, Cheema U. The next level of 3D tumour models: immunocompetence. Drug Discov Today. 2016 21(9):1421-8.