Title: Precision Oncology: A novel approach to modelling neuroblastoma and specific treatment as its underlying genetic abnormalities changes during a patients treatment
Funder: Health and Care Research Wales (Welsh Government)

Abstract: The impact of cancer in Welsh children, their families and communities is significant; progress in cancer outcomes in children has not generally improved over the last 30 years. Each year in England and Wales approximately 85 children are diagnosed with a neuroblastoma (NB). The incidence is 1 case per 100,000 children. In Wales 2-3 children with NB undergo surgery, however only 30-40 % of children with metastatic or relapsed NB survive at 5 years. Despite the low incidence of cases per year – neuroblastoma is a huge burden on paediatric oncological services due to its poor outcome. NB is the most common extracranial tumour of children and is the commonest cancer in infants. NB patients are stratified into low-, intermediate- and high-risk groups, based on different tumour features, including tumor histology, clinical stage, cell ploidy, and MYCN oncogene amplification. Patients with non-amplified MYCN oncogene tumours show overall survival more than 90%. However, the other half of the patients constitutes a high-risk group and these patients harbor MYCN amplification. Our research proposal offers and opportuntity to change the narrative on NB. The vision here is to establish a transformative approach in the treatment of rare paediatric cancers by incorporating molecular profiling, transcriptomics and translating this into an intervention pipeline that targets the tumour microenvironment. The project will utilise the facilities that HCRW has invested in already: Wales Cancer Bank, Wales Gene Park and Wales Cancer Research Centre. The principal aim is to determine the fundamental tumour microenviroment (TME) signatures of neuroblastoma tumours, that enables the development of a reproducible model for neuroblastoma that is patient specific. The long term goal is to use this model to rationalise specific tumour treatment to target specific tumour features. And to monitor tumour status using a liquid biopsy approach. The goals of the project are: 1] We will profile the TME signatures of neuroblastoma to assess the underlying transcriptomic heterogeneity, utilising biopsy samples from archived material in the CCLG tissue bank*. This will be undertaken in MYCN amplified and non-amplified tumours. Followed by validation studies to seek the spatial context of TME biomarkers. 2] Next we will undertake a study to show the origin of chemo-resistance and the acquired presence of a cancer stemness and differentiated stroma in the post-treatment adapted NB tumours. This will be initially undertaken using biopsy material pre and post chemotherapy treatment, providing a means to propose hypotheses in the subsequent organoid model 3] We will establish NB organoid optimisation for the first time in Wales. To develop a novel preclinical model for NB exhibiting self-renewal property and allowing us to obtain a reservoir of NB patients’ biological material useful for the study of NB molecular pathogenesis and to test drugs for precision treatments. 4] Finally, the in vivo organoid model will provide a model system to derive NB specific – extracellular vesicles. These exosomes will be profiled against the organoid culture of origin. This provides the pump-priming evidence that EV liquid biopsies can be used to temporally track tumour status, and treatment responses in patients. *CCLG Tissue Bank is The National Tissue Bank Resource for Childhood Cancers, Newcastle

Lay abstract: Neuroblastoma is the most common tumour of children (other than brain cancers) and is the commonest cancer in infants. Whilst it represents only 8% of all childhood cancers it is a serious disease, responsible for 15% of all childhood cancer deaths. The majority of children present with disease that has already spread, and in this situation fewer than half of them will survive despite aggressive surgery and complex chemotherapy treatments. In fact the treatments we offer to children has changed very little in the past 20-years, and are rather brutal. The chemotherapy drugs are old fashioned and very toxic, and fails more often than not. Despite the fact this is a rare disease, dealing with it is a big burden on cancer services due to its poor outcome, and has a devastating impact on families and healthcare staff. Currently there is very little research into this disease nationally and nothing in Wales, and hence our new initiative brings surgical expertise and basic biological scientists in Wales together to try and address the many unknown questions. Our ultimate ambition is to improve the poor survival outcomes in neuroblastoma. Our proposal aims to understand some of the cellular and molecular changes that are important in dictating how dangerous the disease is. We have experience in measuring RNA- taken from the tumour to establish a “fingerprint” for the disease. We will identify which RNA-factors are different when comparing dangerous to non-dangerous Neuroblastoma. Currently the presence of a specific genetic change (MYCN amplification) is the standard means of doing this- but this tells us little about the complexity of the tissue and other elements that are critical to promoting severe, difficult to treat disease. We are also keen to understand how treatment forces the tumour to change, and adapt to treatment, so we can identify new opportunities for other drugs if/when the disease might return. An exciting aspect of this , is the development of mini-tumours in the test tube. Such “organoids” are well established in Wales for other tumours, but not in this rare disease setting. Establishing organoids will be extremely valuable. Collaborators in Italy will help us achieve this. These test-tube tools will represent the individual and unique aspect of each child’s tumour- and allow us ultimately to explore the use of novel drugs for these tumours in a patient tailored fashion. We have incorporated the HCRW funded Wales Gene Park and WCRC into the study, and plan to recruit two lay research partners (e.g. parents of children with neuroblastoma), to consult on communication strategies. They would help evaluate materials for patient information and consent, and for public engagement activities. Our research findings will be shared to the research community through presentation at conferences and through publication at the conclusions of this project, as well as hosting a workshop which will be accessible to researchers and the public. We have already jointly given a talk at the Rare Cancer Forum (in September 2020) to highlight future intentions in this important and neglected research area, and will contribute to many such events as we proceed.

Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS