Neuroblastoma is a cancer of the nervous system that primarily affects children aged 5 and younger. Although neuroblastoma accounts for only 5% of childhood cancers, it is responsible for approximately 15% of childhood cancer deaths. For children with high-risk neuroblastoma – children in which cancer has spread significantly – the outlook is extremely poor. Approximately 1 in 5 of these children will not respond to treatment, and of those that do, 50% will develop drug resistance leading, in many cases, to death.
Dr Olga Piskareva, an NCRC supported scientist and Honorary Lecturer at RCSI, has recently published a study describing a new way to grow cancer cells in the lab. Traditionally, researchers grow cancer cells in the flasks on the flat surface. This is not the way cells grow in the human body. Dr Piskareva in collaboration with Dr Curtin and Prof O’Brien has designed a new way to grow cancer cells that recreate their growth in 3 dimensions as in the human or mice body. They used special cotton wool like sponges as a new home for cancer cells and populated them with cancer cells. At the next step, they gave cells the drug at the different amount and checked what happened. In this system, cells responded only to the drug at doses used in the clinic or mice 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.
The main challenge in treating high-risk neuroblastoma is to combat tumour metastasis and development of resistance to multiple chemotherapeutic drugs. In the native tissue, cancer cells are surrounded by a three-dimensional (3D) microenvironment which provides biological and physical support and determines disease initiation, progression, patient prognosis and response to treatment. The conventional two-dimensional (2D) cell culture lacks this feature resulting in discrepancies between in vitro and in vivo results. Current neuroblastoma studies employ either 2D cell culture systems or murine models or alternatively a mix of both.
In collaboration with Dr Caroline Curtin and Prof Fergal O’Biren (TERG), we decided to bridge the gap between 2D culture and in vivo tumours in neuroblastoma research by developing a tissue-engineered cell culture model of neuroblastoma. This project is supported by a pilot grant from Neuroblastoma UK.
To understand what signalling pathways are activated in 2D, 3D and in vivo neuroblastoma models, we decided to look closer at the differences between conventional 2D neuroblastoma cells and their xenografts. This way we hope to find those targets that are activated in both tumour microenvironment and the 3D tissue engineered models. Ciara and Larissa have begun this search by profiling xenograft samples with a panel of antibodies. Ciara became particularly fascinated by the elevated levels of c-jun, TCF1 and LEF1 in cisplatin-resistant neuroblastoma xenografts suggesting that the development of cisplatin resistance in neuroblastoma may be accompanied by activation of the wnt/b-catenin pathway in vivo. Larissa identified that cisplatin-resistant neuroblastoma cells secrete chromogranin A (CgA) at levels higher that cisplatin-sensitive cells. CgA levels also correlated with increased vascularisation and volume of murine orthotopic neuroblastoma xenografts. Altogether it suggests that CgA can be used as a marker of neuroblastoma cell growth both in vitro and in vivo.
Irish Association for Cancer Research – Annual Meeting takes place at Newpark Hotel, Kilkenny on Thursday 23 and Friday 24 February 2017.
MCT cancer researchers secured oral presentations at different sessions. Prof Ray Stallings is a guest speaker at the Plenary Session focused on challenges in childhood cancers. He will be discussing ‘Modulation of neuroblastoma phenotype with epigenetically regulated miRNAs’.
Stephanie Annett will be giving a talk ‘FKBPL as a novel prognostic biomarker and therapeutic agent in high-grade serous ovarian cancer’ at Proffered Paper Session on Thursday morning. Two Irish Cancer Society funded PhD students will be discussing their findings at the Irish Cancer Society Scholar and Fellow Presentation session. Louise Walsh – ‘RNA sequencing identifies bromodomain proteins as a therapeutic strategy for invasive lobular carcinoma’ and Brian Mooney – ‘Expression of the cocaine- and amphetamine-regulated transcript recruits BAF chromatin remodelling complexes to the estrogen receptor’.
Neuroblastoma is a childhood cancer caused by the abnormal growth and development of neural crest cells (1). The disease commonly affects children age 5 years or younger. Approximately 50% of children have cancer cells that have migrated to distant sites in the body and formed tumour masses at the time of diagnosis. The main challenge in treating neuroblastoma is to combat tumour metastasis and development of resistance to multiple chemotherapeutic drugs. Despite major advances in available therapies, children with drug resistant and/or recurrent neuroblastoma have a dismal outlook with 5 year survival rates of less than 20%.
Research of Prof. Stallings lab is focused on elucidating the molecular events that contribute to the development and progression of neuroblastoma (2). A major area of research involves the identification and functional analysis of microRNAs that contribute to chemotherapy resistance in neuroblastoma, along with the development of microRNA-mediated therapeutics.
The main research projects were presented at the Departmental meeting on December, 5th.
The first talk by Olga Piskareva has explored how current concepts of development of drug resistant, tumour microenvironment and cell-to-cell communication can be applied to reconstruct relapsed or drug resistant neuroblastoma microenvironment using 3D tumour models.
The second talk was presented by Ciara Fallon. Ciara is our StAR PhD student. She has selected the project ‘Exosome mediated drug resistance in high-risk neuroblastoma’ as her first choice. At the moment she is doing her lab placement in Cancer Genetics group as a part of the RCSI StAR PhD Programme. Built upon results of the former BioAt PhD Student Ross Conlon (3), Ciara’s project is focused on the validation of exosomal miR-548d-5p as a regulator of cell viability and proliferation in cisplatin sensitive and resistant neuroblastoma cell lines.
Finally, the last, but not least was a talk by John Nolan. His talk entitled “MiRNA-124-3p Reduces Cell Viability in Cisplatin Resistant Neuroblastoma Cell Models” was focused on the results submitted to the Royal College of Surgeons for the Degree of Doctor of Philosophy. His studies cover the development of cross resistance to other drugs, investigation of common altered proteins and signaling pathways in cisplatin resistant neuroblastoma cell lines and validation of miRNA that can target these proteins and stop cell proliferation. Part of the results was published last year in Cancer Letters (4).
The work carried out in Prof. Stallings lab is supported through the research grant to Prof. Ray Stallings and PhD fellowship to John Nolan by National Children’s Research Centre, Crumlin Hospital.
Davidoff, A. M. Neuroblastoma. 2012 Semin. Pediatr. Surg.21, 2–14.