Manipulating NF-kappa oscillations and linked pathways to promote cell death in neuroblastoma primary cultures and cel

Neuroblastoma is a cancer with a dismal prognosis for very many affected children. Very occasionally, a neuroblastoma tumour disappears spontaneously. This has directed researchers to investigate mechanisms that might encourage all types of neuroblastoma to similarly ‘self destruct‘.
N-Myc is a protein strongly associated with resistant neuroblastoma; NF-?B proteins are also known to control neuroblastoma behaviour: manipulating these two ‘molecular switches‘ may aid tumour destruction. For example our initial studies indicate that persistent ‘cycling‘ of NF-?B proteins helps neuroblastoma cells persist.
Retinoic acid (RA) derivatives are used with limited success as part of therapy for resistant neuroblastoma. We propose that RA may work better with anti-cancer drugs by altering these molecular switches in resistant neuroblastoma cells. Ultimately, this work aims to make current neuroblastoma treatment more effective. Unusually, our work will benefit from the unique opportunity to study neuroblastoma cells grown directly from resistant tumours in our laboratory.
This project and the multidisciplinary expertise it harnesses represent a great opportunity to transform the outlook of this devastating childhood cancer.

Neuroblastoma, the most common extra cranial solid tumour in childhood, is associated with incorrect regulation of neuronal apoptosis. I aim to characterize the role of key pathways that regulate apoptosis, cell proliferation and differentiation in neuroblastoma cells. The pathways of interest include NF-?B, p53 and N-Myc. NF-?B and N-Myc have both been implicated as being involved in regulation of proliferation and cell death in neuroblastoma. The cell differentiation agent Retinoic acid (RA) has been used clinically to block neuroblastoma proliferation. Interestingly, this agent modulates the activity of both N-Myc and NF-?B.
Pioneering work in the Centre for Cell Imaging (CCI) has developed state-of-the-art expertise and facilities for single cell-based assays for multiparameter measurement of signalling dynamics, transcription and cell fate. This group previously showed that deregulated NF-?B in neuroblastoma cells results in sustained nuclear-cytoplasmic oscillations in NF-?B localization leading to prolonged NF-?B-dependent gene expression.
In this Fellowship, I propose to apply this expertise to study the regulation of cell fate in neuroblastoma cells. The aims of the research will be to investigate: 1) The regulation of NF-?B signalling in neuroblastoma cells and its relationship to cell death; 2) The effects on cell fate of manipulation of N-Myc and NF-?B signalling using chemical and genetic inhibitors; 3) The effects of treatment with RA and clinically used anti-cancer agents and their combination with inhibition of the signalling pathways.
This work will benefit from the derivation of large scale cell cultures from primary tumours which has recently been successfully achieved in the CCI in collaboration with the surgery team at Alder Hey Children‘s Hospital. Cells from several of these tumours have been cultured and frozen down in large stocks and a protocol for this procedure will be applied for culture of cells from all future neuroblastomas at Alder Hey Hospital where consent for research use is given. This will ensure that the study maintains relevance to patient disease.
This work benefits from an established collaboration between the CCI and Alder Hey Children‘s Hospital. Monthly combined meetings alternate between sites to review ongoing work and comprises cell biologists (White and Spiller), a BBSRC-funded neurobiologist (See), academic paediatric surgeons (Losty and Jesudason) and paediatric oncologists (McDowell and Pizer). This provides an excellent multidisciplinary environment for an MRC Clinical Training Fellowship.