#childhoodcancer – Page 3 – Blog about neuroblastoma research

First Research Meeting For A Medical Student

At the beginning of my career, I worked for two years in a Ukrainian company organizing international industrial conferences. So I have insider knowledge of how the conference works, and that the determining factor for the success is the active communication between the participants. And at the RCSI research day and Cork IACR conference, this component was perfect. At both events, I presented my poster and had a chance to discuss the recent advances in neuroblastoma epigenetic drug research. During RCSI Research day, I was excited to learn about the accomplishments of other undergraduate studies and was thrilled to learn that my classmate is participating in research too. He had developed an online recourse to practice cardiac auscultation, which is extremely useful for my medical studies. But professionally, I enjoyed the cancer research posters and presentations at the IACR conference and was eager to meet the researchers working on medulloblastoma, a paediatric neural cell cancer, and the research team from UCD, the neighbours of our university who worked on breast cancer. It was the most valuable opportunity to take a glimpse into other research, become inspired by the most ingenious methods, and cultivate professional knowledge and personal connections – I am so lucky I have been at RCSI Research day and the IACR conference! I have greatly enjoyed my time, and I am looking forward to (hopefully) going to the next year’s conferences again.

Written by Nadiya Bayeva

RCSI Research Day 2022

Cancer Bioengineering Group thoroughly enjoyed getting back to in-person Research Day at RCSI after 2 years, we’re now very much looking forward to the IACR conference later this month! We will have 2 oral and 5 poster presentations at IACR 2022.

Ronja: My typical day

A typical day for me is difficult to describe because there are many facets to a PhD in the Cancer Bioengineering research group. Some days I spend in the lab sectioning, staining or looking at tumour samples under the microscope. Others I stay at home, read papers and try to figure out how they can help me to achieve my research goals. Some days I take part in the courses and workshops offered in the scope of a structural PhD. Then there are times when I sit here writing up for you guys what it is that I do those other days. The academic environment also provides lots of other opportunities to apply yourself and broaden your horizons or pursue what you enjoy. I, for example, have the chance to partake in weekly dissections for medical teaching which helps to keep my anatomical knowledge fresh and is an always welcome change of scenery (and smell) when I am stuck on other things. Furthermore, I get to see the other side of conferences and what is involved in their planning, because I am part of the local organising committee for the European Federation for Experimental Morphology Symposium 2022.

Figure 1 Working on neuroblastoma cancer the samples I am working with are quite unsurprisingly tumour cells. But these can be grown, for example, in mice (A) or on manmade scaffolds (D). I am moving a staining rack that holds the microscopy slides through staining containers filled with different solutions (C) to stain the slides. After the slides are stained the excess stain is removed by washing in distilled water (B). The resulting images depend on the type of stain. Stains like Alcian Blue can only be viewed with brightfield microscopy (A). But Picrosirius red can also be viewed under polarised light or as seen here (D) with fluorescent microscopy.

Currently, not yet half a year into my PhD, a lot of my time is spent planning. That’s planning which methods to use, which products to order and which experiments, and analyses would result in the most coherent and rounded off story being told by the summation of my research. I also spend a lot of time optimising the methods I will use to assure reproducibility and avoid issues during the analysis later on. For example, the whole tumour sample stained with Alcian blue you can see in Figure 1A clearly shows discernible blue and red regions. However, I have spent about 2 months now trying to get to a point of producing this same outcome reliably rather than having samples show up entirely blue or very only faintly stained. Picrosirius red, the solution I used to stain the sample in Figure 1D stains collagen. But there are many different stains for collagen. After researching most if not all of them I chose this one because it can be viewed with different types of microscopies providing slightly different information. Another step of planning includes how many pictures of which magnification will be required, one image of a whole section for orientation such as in Figure 1A and then more zoomed-in images to investigate the structure of collagen such as in Figure 1D.

Between course work and planning and optimising different aspects of my project, my PhD provides me with plenty of opportunities to focus on something else whenever I get stuck to later return with a fresh set of eyes.

Written by Ronja Struck, a 1st Yr PhD student funded by the IRC-CFNCRF

Welcome to the Cancer Bioengineering Group!

It is time for a full group presentation here at the blog! Throughout the month we shared about our group members and their research focus on Twitter. Now, we would like to share more about the group here and invite you to keep following us on social media.

The Cancer BioEngineering Group is a research group led by Dr Olga Piskareva at the Royal College of Surgeons in Ireland. The group has 6 PhD students developing research projects around neuroblastoma biology.

Our projects address topics related to neuroblastoma microenvironment, cell interactions, tumour resistance and the development of new therapies. To do that we use 3D in vitro models, identify immunotherapeutic targets and evaluate extracellular vesicles.

We are a dynamic group proud to be engaged in research, science communication and patient involvement. We do that through different initiatives.

We support and collaborate with several neuroblastoma charities around Ireland and internationally such as the Conor Foley Neuroblastoma Foundation, the National Children Research Centre, the Children’s Health Foundation Crumlin and the Neuroblastoma UK. Moreover, our projects are funded by the Irish Research Council in partnership with these charities and by RCSI StAR PhD programmes.

We promote neuroblastoma awareness through different activities. For instance, last September at the Childhood Cancer Awareness month we promoted a hiking challenge to raise money and increase awareness of neuroblastoma. We hiked for 30km at Wicklow mountains in a day and raised over € 2,000 for neuroblastoma research charities.

We are also present in social media, creating content in the form of blog posts and tweets to share the science we are doing.

We are always happy to answer questions and interact with the public. Follow us on our social media channels and read our blog to know more about us and our research.

Thanks for reading and we go ahead with neuroblastoma research!

Written by Luiza Erthal

#AskLuiza: How Does The Microenvironment Influence Neuroblastoma Cells?

Understanding how tumour cells interact with the other cells in the body is crucial for an effective treatment. Moreover, it can help to identify patterns that are exclusive of tumour cells to be a target in treatment.

The interactions of tumour cells with the surrounding tissue, the microenvironment, affects chemotherapy sensitivity, immune cells recognition and expression of molecules on the cell surface, to only cite a few interferences.

This is particularly crucial in metastatic cells, which are cells that have spread to other parts of the body coming from the primary tumour location. Specifically, for neuroblastoma half of patients with high-risk disease present a metastatic tumour at the diagnosis. In addition, one of the organs that are mostly populated by metastatic neuroblastoma cells is the bone marrow.

A review paper recently published address some important aspects about the interactions between neuroblastoma cells, bone and bone marrow resident cells¹. This review argues in favour of understanding these interactions to search for new targets for therapy.

However, neuroblastoma cells proved to be difficult to characterise due to dynamic changes induced by external stimuli. Therefore, neuroblastoma cells change upon exposure to the bone marrow microenvironment.

The authors present some studies showing that neuroblastoma cells infiltrating the bone marrow express receptors for small proteins called chemokines that induce cell adhesion in the bone marrow. On the contrary, the cells did not present on their surface molecules that stimulate the immune system recognition. Therefore, they are naturally invisible to the action of this system.

Moreover, it has been shown that metastatic tumour cells release extracellular vesicles expressing GD2. These vesicles have an important role in cell-cell communication and the GD2 is a marker exclusive of neuroblastoma cells. Thus, it facilitates the identification of metastatic cells.

These alterations on neuroblastoma cells surface after they interact with bone marrow cells may facilitate the invasion and spread of the tumour. Thus, looking closely to that may help to develop more effective treatments for neuroblastoma.

At the Cancer Bioengineering Research Group, many of our projects are related to tumour resistance, cell interaction and the tumour microenvironment. These three aspects are very important to understand neuroblastoma at the tissue level. We study them and expand this research to applied projects aiming at the development of new therapeutic modalities.

For instance, we are currently evaluating the effect of extracellular vesicles from different neuroblastoma cell lines in the induction of proliferation and increased viability. Moreover, we are studying the interaction of neuroblastoma cells with immune cells such as macrophages. Finally, we are also identifying targets to develop an anti-tumour nucleic acid-based vaccine against neuroblastoma.

We go from basic to applied research interconnecting the findings and expanding the understanding of neuroblastoma biology. Ultimately, we aim to improve treatment and quality of life for patients.

Written by Luiza Erthal

References

1. Brignole, C. et al. Bone Marrow Environment in Metastatic Neuroblastoma. Cancers 13, 2467 (2021).

#AskLuiza: What are the main differences between cancers in adults and children?

Looking carefully we can easily see that children are very different from adults. They have different needs, desires, likes and dislikes. Not surprisingly, the children body is also very different in their functioning and response to medical needs. Therefore, cancer in children has many different characteristics when compared to cancer in adults. Childhood cancer is different in terms of the most common types, the causes, the treatment and the course of the disease.

Firstly, childhood cancer is rare and this sometimes impairs an early diagnosis. Therefore more aggressive diseases tend to be present at the time of diagnosis. Nevertheless, there are specific types of cancer that are more common in children, which helps in the diagnosis. They are cancers affecting the blood and lymph nodes (leukaemia and lymphoma), the brain (astrocytoma), the liver and the bones (osteosarcoma). These types of cancer are less common in adults.

Another important difference between adult and childhood cancer is the leading cause of the disease. Most of the time the cause of childhood cancer is unknown, although genetic contributions related to overexpression or deletion of genes can be determined. On the other hand, adult cancers are frequently associated with alterations in the DNA (mutations) as well as lifestyle.

The treatment plays an important role in the differences between adult and childhood cancers. Usually, similar treatments are used for both adults and children, including chemotherapy, radiotherapy, surgery, transplants and immune therapy, according to the type of cancer and its stage. However, the doses and types of drugs may differ between them. The differences in the treatment go beyond the doses and encompass the mechanisms of action and possible long term toxicities of drugs. For example, the use of drugs that damage DNA can be prohibitive in children due to the increased risk of secondary cancers in the future.

In conclusion, specific types of cancer are more common in children and the cause of this disease is frequently unknown. Fortunately, children have great possibilities to survive cancers but the treatment needs to be carefully chosen and its long-term effect on the body have to be monitored for their whole life.

Written by Luiza Erthal

References

Kattner, P. et al. Compare and contrast: pediatric cancer versus adult malignancies. CancerMetastasis Rev. 38, 673–682 (2019).

How Childhood Cancers Differ From Adult Cancers. Available at https://www.winchesterhospital.org/health-library/article?id=30409

Accessed November 18, 2021.

How childhood cancers are different from adult cancers. Available at https://medlineplus.gov/ency/patientinstructions/000845.htm

Accessed November 18, 2021.

How is Childhood Cancer Different from Adult Cancer? Available at https://www.acco.org/blog/childhood-cancer-differs-from-adult-cancer/

Accessed November 18, 2021.

#AskLuiza: How is the neuroblastoma stage determined and how does this impact treatment?

The determination of the tumour stage is an important step after a neuroblastoma diagnosis. The stage of neuroblastoma is determined depending on tumour location and if it has spread to other parts of the body. This will guide risk group assignment and treatment choice.

The first staging system for neuroblastoma, the International Neuroblastoma Staging System (INSS), was developed in 1986 and is based on the pathological evaluation of the tumour after a removal surgery. In 2005, The International Neuroblastoma Risk Group Staging System (INRGSS) started to be used. This system is based on tumour images before any surgery. Therefore, it is based on image-defined risk factors to determine the tumour stage (see table below). It also uses clinical, pathologic, and genetic markers to determine the risk groups, which can be low-risk, intermediate-risk, or high-risk.

Reference: Neuroblastoma – Childhood: Stages and Groups, Cancer.net.

Recently, the Children’s Oncology Group (COG), a clinical trial group dedicated to paediatric cancer research revised the classification system they use to determine tumour stage for enrolment in clinical trials¹. Previously, they have been defining the tumour stage based on the INNS system. Now they proposed a revised classification that takes into account the INRGSS and chromosomal alterations.

Key clinical and biological factors used in the neuroblastoma risk classification include age at diagnosis, disease stage, tumour tissue appearance under a microscope (histology), the status of the gene MYCN that affects tumour growth, the amount of DNA in a tumour cell (called tumour cell ploidy), and alterations in the DNA.

They analyse the outcome of almost 5,000 patients to define risk groups based on the INRGSS, using alterations in the DNA of tumour cells as a biomarker and considering current therapy modalities. In general, they found that the correlation of stages between systems is not exact. However, the differences in survival were minimal when comparing staging systems, which corroborates the use of the revised version.

In general, the new version classifies L1 and L2 tumours as low risk, except for L1 tumours with alteration in the gene MYCN and that cannot be removed by surgery, which is high-risk. For L2 tumours, MYCN status and age can be used to evaluate prognosis. Stage M tumours can be classified as high risk or intermediate-risk depend on age, MYCN status and DNA alterations. In conclusion, low-risk groups have excellent outcomes with any or limited therapy, the intermediate-risk group have very good outcomes and high-risk groups have inferior outcomes despite therapy.

This new version of the COG classifier will provide a uniformization of patient risk classification for clinical trials, ultimately enabling the comparison between different trials.

Written by Luiza Erthal

Reference:

1. Irwin, M. S. et al. Revised Neuroblastoma Risk Classification System: A Report From the Children’s Oncology Group. J. Clin. Oncol. JCO.21.00278 (2021)

#AskLuiza: Could we use blood tests to detect neuroblastoma?

For most neuroblastoma cases a tissue biopsy, which means remove a piece of the tumour and analyse under a microscope, is performed to confirm the diagnosis, allow the risk stratification (low, intermediate or high-risk cancer) and determine treatment. However, it is not always possible to perform a biopsy. In these cases, other tests are available such as ultrasound, x-ray and urine test¹. Although these tests help the diagnostic, they are much less informative to determine the risk group and to guide treatment.

Cancer is a genetic disease and as such several genetic alterations are present in the DNA and can be useful to determine a diagnostic and prognostic of the disease. These alterations can also be useful to monitor treatment effects. But, how we can examine the DNA without using a piece of the tumour?

The answer for that relies on our blood. Cancer patients have cancer cells circulating in their blood and these cells release what we call cell-free tumour DNA. The level of cell-free tumour DNA in the blood of a healthy individual is low while this is increased in cancer patients. The detection of cell-free tumour DNA with specific alterations may help to not only detect the disease but also determine if the tumour cells are changing after treatment.

Recently, a trial for a blood test that can detect 50 different types of cancer was launched in the UK². The test called the Galleri test look for cell-free tumour DNA in the blood using modern genetic sequencing technology. It spots the DNA that has changes common in specific cancers but not seen in healthy cells.

One of the main questions of this trial is if the test can find early stages of cancers. Although neuroblastoma is cancer that could benefit from this test, unfortunately, it is not one of the cancer types detected. However, the presence of cell-free tumour DNA was already detected in neuroblastoma patients’ blood³. Moreover, several alterations in the DNA of neuroblastoma patients have already been reported to have predictive value for disease progression and treatment monitoring. These alterations include the increase in the number of specific genes, genes breakdown or increased activity of certain genes. Their presence may indicate poor outcomes and early detection could guide to specific treatment options.

The big challenge is to have a non-invasive diagnostic method, such as blood tests, that are sensitive enough to detect the early stages of the disease. Specifically for neuroblastoma, comprehensive analysis in clinical trials regarding cell-free DNA levels and their specific changes over time would help to advance the development of liquid biopsies, such as blood tests, for this type of tumour.

Written by Luiza Erthal

References

1. Tests for neuroblastoma, Cancer Research UK. February 25, 2021

2. The Galleri multi-cancer blood test: What you need to know, Harry Jenkins, Cancer Research UK. September 13, 2021.

3. Wei, M., Ye, M., Dong, K. & Dong, R. Circulating tumor DNA in neuroblastoma. Pediatr. Blood Cancer 67, (2020).

#AskLuiza about neuroblastoma biology

#AskLuiza

We are launching a new initiative #AskLuiza to help the public and patients know more about advances and current trends in neuroblastoma.

Luiza is a research writer at the Cancer Bioengineering Research Group. She holds a PhD in Biomedical Sciences from Trinity College Dublin. You will ask a question and Luiza will look for the answer in peer-reviewed research papers that the research community trust.

Leave your question and follow our blog to read the answer soon: https://forms.gle/vrgwKoZivyUVawk9A

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.

*Credits for the image: 3D models to study neuroblastoma. Adapted from Nolan, J. C. et al. Preclinical models for neuroblastoma: Advances and challenges. Cancer Lett. 474, 53–62 (2020).*

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.