One of my classmates shared a funny image of the Upturned Microscope and I could not agree more… That week was exactly 12 months since I have started to apply for funding to carry out neuroblastoma research. Seven applications were submitted to various national and European funding bodies. Some were for a PhD student project, other were for PostDoc or both, Four came back with nothing. Two – decision pending and another one was shortlisted but not funded in the first round. A possibility existed that it could be funded at some day in 2017. So, this pic illustrated my feelings very well..
However, either my efforts were paid off or some Christmas miracle has happened, and I have received a note informing on the green light to funding my application. The funding comes from the National Children’s Research Centre and covers a PostDoc position and research consumables over 3 years. It was the very Christmasy news! I was over the moon.
The conditions of the award states:
Finally, the applicants must indicate how the investment in this research will be measured and communicated, not only to the academic community but also to the hospital, the fundraisers, to parents and patients and to the wider public.
This place will be my research diary to share ups and downs of the ongoing work. Please join me in this exciting journey!
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.
I am a medical student at Penang Medical College under a twinning programme with the Royal College of Surgeons in Ireland. I studied my pre-clinical years at RCSI Dublin. In the summer of 2015, I had the opportunity to join the RCSI Research Summer School (RSS) Programme. I was mentored by Dr Olga Piskareva, from Cancer Genetics, Molecular and Cellular Therapeutics (MCT) Department, RCSI. Being in this lab was simply one of the greatest experiences I have in my life; it was really rewarding.
My RSS project investigated the role of VDAC-1 protein on chemotherapy resistance in neuroblastoma. The only research focus of this lab is to find key players in neuroblastoma pathogenesis and to advance anti-cancer therapy.
Neuroblastoma cell line SK-N-AS. The cell line in my experiments.
I was entrusted with the task of splitting cells. I would plate them onto 96-well plates, add cisplatin drug and measure their viability afterwards. It may sound simple here, but the whole process required passion and hard-work.
Prior to this, I did not have any experience in the medical research field. During my first two weeks, everything seemed so tough; however, they became easier as the weeks flew by. My mentor, Olga, and the other staff and PhD students (Garret, John and Ross) were helpful and always guided me to explore my potentials. This programme taught me various new things which I would not have acquired on a normal day-to-day basis in school.
Introduction to Malaysian cuisine.
The people at Cancer Genetics were warm and wonderful. The hospitality, love and guidance cannot be quantified and words cannot express my immense gratitude towards them. It has been fascinating and I cherish every moment I spent there. We bonded over our weekly breakfast and tea sessions so well, and I am indeed grateful for being a part of this big family. It is my sincere wish that this positive spirit of togetherness will be preserved and will grow stronger in the future. This is something special, and I think ours is the best lab at RCSI!
I returned to Penang Medical College to further my studies in my clinical years. I took part in the PMC Research Day 2016 in which I was awarded the First Prize in Oral Presentation. I would like to dedicate this success to Olga and everyone who has been with me throughout my time at Cancer Genetics. Without all the guidance, I would not have made it this far.
I strongly urge students to take part in the research opportunities, because you gain invaluable experiences that you do not get elsewhere. May whatever we do at the lab today make a difference in another person’s life someday in the future.
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.
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:
Schweiger PJ, Jensen KB.Modeling human disease using organotypic cultures. Curr Opin Cell Biol. 2016 43:22-29.
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.
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.
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.
For children who do survive cancer, the battle is rarely over. Over 60% of long‐term childhood cancer survivors have a chronic illness as a consequence of the treatment they received; over 25% have a severe or life‐ threatening illness. How much do we know about quality of life of childhood cancer survivors?
Researchers in health- and illness-related social sciences understand that the there is a life after the treatment completed. The life is full if diverse levels and issues from health related to social adaptation in different shapes and forms. Children and teenagers may experience fear when returning to school due to temporary or permanent changes to their physical appearance (1,2). They worry about their ability to socialise with their friends due to lengthy absences (3–5). Treatment can result in the development of learning disabilities in children and thus marking school as a major source of frustration (1,2). These learning difficulties can affect a child’s confidence and self-esteem, if left without attention and care (1,3). All studies come to the same conclusion. Challenges in education of children with cancer are complex, however most can be tackled efficiently through planning and good communication (1–5).
It is important not only to recognise the problems but to start changing the situation. Apparently much more could be done more efficiently if patients are involved in setting up future research agenda.
Reading
Gurney JG, Krull KR, Kadan-Lottick N, Nicholson HS, Nathan PC, Zebrack B, et al. Social outcomes in the childhood cancer survivor study cohort. J Clin Oncol. 2009;27(14):2390–5.
McDougall J, Tsonis M. Quality of life in survivors of childhood cancer: A systematic review of the literature (2001-2008). Supportive Care in Cancer. 2009. p. 1231–46.
Barrera M, Shaw AK, Speechley KN, Maunsell E, Pogany L. Educational and social late effects of childhood cancer and related clinical, personal and familial characteristics. Cancer. 2005;104(8):1751–60.
Langeveld NE, Stam H, Grootenhuis MA, Last BF. Quality of life in young adult survivors of childhood cancer. Support Care Cancer. 2002;10(8):579–600.
Klassen AF, Anthony SJ, Khan A, Sung L, Klaassen R. Identifying determinants of quality of life of children with cancer and childhood cancer survivors: A systematic review. Support Care Cancer. 2011;19(9):1275–87.
Yeh JM, Hanmer J, Ward ZJ, Leisenring WM, Armstrong GT, Hudson MM, et al. Chronic Conditions and Utility-Based Health-Related Quality of Life in Adult Childhood Cancer Survivors. J Natl Cancer Inst [Internet]. 2016;108(9):4–7.
Armstrong GT, Chen Y, Yasui Y, Leisenring W, Gibson TM, Mertens AC, et al. Reduction in Late Mortality among 5-Year Survivors of Childhood Cancer. N Engl J Med. 2016;374(9):833–42.
Ok. Now, when the stress of the presentation is over, I am happy to share new technologies used during the SIOP2016. As I mentioned yesterday, my work was selected for e-poster presentation. It looked this way:
This is e-poster station, where anyone can look up all posters displayed during the meeting.
It is definitely a step forward. Anyone can look up any poster, listen to a commentary recorded by the author, zoom in and out and send a request/comment to the author. It looks cool and trendy. Though, you can feel invisible as no physical copy displayed in a designated area. No crowds of poster presenters and judges. No waiting faces desperate to share their study…
The actual Poster Discussion session was a traditional presentation when my poster was up on the big screen, I had 8 minutes to convince the audience navigating through figures. This session was late and no many attendees survived to come and challenge your statements. Nevertheless, it was enjoyable experience. 🙂
SIOP is the International Society of Paediatric Oncology. It is a global multidisciplinary society representing doctors, nurses, other health care professionals, scientists and patients or their relatives. The Society’s motto is ‘no child should die of cancer’. The meeting 2017 is being held in Dublin, the city where I live and work.
Indeed, it was appealing to attend the key meeting in childhood oncology field. As any participant, I had an opportunity to submit an abstract about my research. To no surprise at all, I received email notifying me on my work being selected for e-Poster presentation. Common stuff. The email also said that it would be displayed at designated stations, like big screens throughout the meeting. Very unusual format, but we are living in the digital technology era; things are changing all the time. So, I would not need to stay by the poster this time. Great – more time for networking and talks.
Then I received another email informing about a Poster Discussion session, which I assumed to be a standard procedure when a group of selected piers stand by your poster and ask Qs. None comes in majority cases. A participant stands and waits and waits till the session is over. So, of course I took it easy.
A day before the meeting, I downloaded the meeting app and started to browse along the content and features. Out of curiosity, I checked details of the Poster Discussion session. This was the moment of mental breakdown – I discovered being selected for an oral poster presentation! My chances were 1 in 1475 (the number of submitted abstracts). I should probably also buy a lottery ticket tonight. Could lucky things come together?
I will reflect on the new e-poster presentation experience later today…
We do not hear much on neuroblastoma or childhood cancer in our everyday life unless we know a child affected by the disease. How much information can we get from newspapers? How do newspapers report it? What is their focus – a child, his/her family or social circles? In the next posts I will try to get insights from the content of newspaper’s stories.
To start with, I needed to select those newspaper articles that cover a story of a child with cancer over a period of time. To do that I selected a recent period from January 1, 2010 to December 31, 2015 because two major events happened during this time.
The first was the 2011 Pulitzer Prize book written by Siddhartha Mukherjee ‘The Emperor of All Maladies: A Biography of Cancer’ which describing the history of both adult and childhood cancer treatment development and research (Mukherjee 2010). Importantly, this book inspired to produce a documentary film of three episodes of two hours each and released in 2015 (Goodman 2015). Researchers in social sciences agree that the publication of a prominent popular science book can lead to increased media interest in the issues raised in the book (e.g. (Nisbet & Fahy 2013).
Another most recent milestone was a breakthrough in childhood cancer management – the FDA approval of a novel drug Unituxin (dinutuximab) for neuroblastoma, the most common solid tumour in children (U.S. Food and Drug Administration 2015).
The timeline of media coverage analysis
next step is actual paper selection. There is an archive of all newspapers and magazines called Nexis®UK database. This database stores copies of all printed papers and magazines worldwide as well as online versions.
Then an a search for articles carried out looking for key words:‘child’, ‘children’, ‘childhood’, ‘kid(s)’, ‘cancer (s)’, ‘tumour (s)’, ‘treatment’, ‘chemotherapy’, ‘radiation’ and their combinations. Only articles published by the UK national newspapers and included key words in either the headline or text were selected. The search returned 255 articles. Of 255 articles, 84 contained a story about a child with cancer. The rest were standardised obituaries in the ‘announcements’ or ‘deaths’ sections, horoscopes containing the word ‘cancer’, fundraising and charity activities not involving an identified child, funding initiatives related to pure cancer research and service, reports on scientific achievements or challenges were excluded.
The selected 84 articles were published by the broad range of local and national papers that are daily on sale through news outlets to a general public in the UK.
Summary of selected newspapers
The Q1: What has been the level of media attention to childhood cancer?
In the last three years, in average 20 articles per year across 9 UK newspapers were published telling us a story about a child with cancer. In other terms it is less than 2 stories per month. It is not enough to raise awareness, educate the public and form their opinion. Tabloid editorial was more interested in this type of stories than broadsheet. This observation is likely due to the nature of tabloid paper interests looking for people personal stories, entertainment, sports and scandal.
Unfortunately, no change in media coverage of childhood cancer was observed around these two milestones (in 2011 and 2015). All together it suggests that neither of them had a strong influence on journalists or editorials regarding childhood cancer coverage.
Frequency of stories about a child with cancer by year (A) and type of newspaper(B)
Interestingly, the growing trend in media coverage of children with cancer occurred by the increase in neuroblastoma coverage. Neuroblastoma had a nearly two fold boost in 2013 vs 2012 and then slightly declined in 2014 and 2015. Nevertheless, the FDA approval did not trigger attention to this cancer.
Frequency of stories about a child with cancer by cancer type
The conference on models and tumour microenvironment has brought together International experts in this field. Two keynote speakers (Peter Friedl, Radboud UMC/MD Anderson and Andrew Ewald, John Hopkins University) presented exhaustive experimental data on plasticity and microenvironmental control of cancer invasion and metastasis.
Their research teams independently found that
Tumour cells migrate collectively as a team from a piece of tumour like a group of people who changed their minds and decided to travel by bus when the majority stayed camping. However, Andrew Ewald acknowledged that they are not pioneers in this discovery. In 1976 Liotta observed migration of tumour cells in a group of 6-10 cells.
A migration group of cells has their leaders who crave the path through surroundings to the new locations.
Leader cells depend on cancer types. It can be any tumour cell in some cancer types or a specialised one.
Migrating cells take shape and follow the pattern of tissues to be invaded.
The experiments by Ewald’s research team on collective cell migration. In short, they co-implanted two lung tumour cell populations labelled differently into mice. One cell population had a green protein tag, another had red. After 6-8 weeks, researchers examined metastases and found that they had a mixed population of green and red tumour cells.
Multicellular seeding is a frequent mechanism for distant metastasis. (A) Schema of multicolor lineage tracing assay. ROSAmT/mG;MMTV-PyMT tumor organoids were treated with adenoviral Cre to induce recombination from membrane tdTomato (mTomato) to membrane eGFP (mGFP). Mosaic tumor organoids were then transplanted into nonfluorescent NSG host mice. After 6–8 wk, lungs of these mice were harvested. If metastases arise exclusively from single-cell seeding, there should be only single color metastases. In contrast, multicellular seeding should produce metastases with both colors. (B) Representative micrographs of polyclonal lung metastases of different sizes. n = 355 polyclonal metastases, across 16 mice and 4 independent experiments. (C) Representative micrograph of a mosaic tumor organoid treated with adeno-Cre and grown in 3D Matrigel with intermixing of red and green tumor cell clones. (D and E) Representative micrographs of primary tumors arising from mosaic tumor organoids transplanted into NSG host mice. Primary tumors varied in their local mixing of red and green tumor cell clones (local mixing %). These differences correlated with the percentage of multicolored metastases detected in the lung (% multicolored). n = 12 mice, 4 independent experiments, 4,072 metastases. Correlation determined by Spearman rank test for samples with more than five lung metastases per mouse. (Scale bars, 20 μm in B and C, and 2 mm in D.)
The very first human cancer cell line was developed from a patient with an aggressive cervical cancer in 1951. This cell line was called HeLa after the patient name – Henrietta Lacks. This is the most popular and robust cancer cell line in biomedical research. Since then, other cancer cell lines were developed including neuroblastoma.
The first successful neuroblastoma ‘cell lines’ were cell populations from tumours that were adapted to grow for a short period in the lab environment in 1947. These tumour cell populations were used as a tool for diagnosis. This success inspired other researchers to develop long-term or immortal neuroblastoma cell lines. To date different neuroblastoma cell lines exist.
Cancer cell lines are sensitive and delicate in handling. They can only grow in the safe environment. Researchers have to protect them against bacteria, low temperatures, and too acidic/alkaline conditions. We protect cancer cells from bacteria contamination by handling them in cabinets where all plastic and media are sterile.
Handling neuroblastoma cells in the cell culture cabinet.
Cancer cells like to grow in conditions similar to conditions in human body. They like temperature of 36.6 – 37C. To achieve it special ‘green cell houses’ – CO2 incubators are built, which maintain the constant temperature, humidity and CO2 concentration.
We place cells in plastic dishes or containers called flasks and keep flasks in the ‘green cell houses’.
The cell growth and well being are checked regularly using microscopes. Healthy cells are to have similar shape, even distribution and grow attached to the plastic surface. Most microscopes have a camera attached to the top and linked to a computer. It helps to take picture of growing cells and record changes in cell behaviour.
Microscopic examination of drug resistant neuroblastoma cells KellyCis83. Cells look healthy and can be kept for another 2-3 days to form a more dense population.
