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UCL Great Ormond Street Institute of Child Health

Great Ormond Street Institute of Child Health

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Design of nanoparticle-based, Toxin Gene Systems For Neuroblastoma Treatment

Supervisors names
Andrew Stoker
Stavros Loukogeorgakis

Project outline
Neuroblastoma (NB) is a paediatric disease of the sympathetic nervous system, accounting for 15% of childhood cancer deaths. High grade tumours remain difficult to treat, with tumour relapse being almost invariably fatal (ref.1). During residual disease therapy patients receive high dose retinoic acid (RA) as a cell differentiation treatment. This improves 5-year overall survival, but only modestly (ref.2). RA induces tumour cell differentiation in culture, but again not completely (ref.3). Thus, RA’s maximal potential as a clinical agent remains unclear. We therefore propose an alternative way of harnessing the clinical potential of RA signalling, by generating a “toxin vector” system (ref.5) which converts the differentiation response to a cytotoxic response in tumour cells. RA amplifies the transcription of target genes and we hypothesise that linking toxin genes to RA-responsive promoters in plasmids should generate a suitable vector for delivery to NB cells. The delivery approach will employ a range of nanoparticles that we aim to target with high specificity to neuroblastoma tumours (ref.5). The student would therefore design and optimise the vectors, examine ways in which to maximise their delivery to tumour cells using nanotechnology and then investigate this toxin gene therapy approach in pre-clinical models.

Aims/Objectives:
The long-term aim is to re-purpose RA as a directly cytotoxic NB therapy.

  • Objective 1 (Month 1-24). To develop an optimised, DNA minicircle-based toxin vector encoding highly RA-responsive toxin genes in cultured tumour cells. The student will use recombinant DNA approaches to construct plasmids encoding luciferase and toxin genes driven by retinoid-responsive promoters. These will be optimised for inducible expression in transfected tumour cell lines, using luciferase and cell survival assays. The vectors will be further modified for use as minicircle DNAs and tested for their expression level and longevity in transfected cultured cells. This objective also includes further development of toxin genes themselves, with use of novel approaches for generating better tumour specificity.
  • Objective 2 (Month 12-24). To optimise our peptide-based liposome technology for targeting a range of NB cells. The student will test the cell targeting ability of existing nanoparticles in NB cell lines and PDX cell lines and will design and test novel, peptide-based and antibody-based targeting strategies for these liposomes to improve targeting efficiency.
  • Objective 3 (Month 12-36). The third objective it to demonstrate a complete toxin gene vector system capable of impeding NB tumour growth in vivo. The student will complete the study by adapting the liposomes for optimal use in vivo (ref.5) and these toxin gene-loaded liposomes will be tested in NB xenograft models and genetic models of neuroblastoma. 


References:

  • Matthay et al. Nature reviews. Disease primers 2, 16078, (2016).
  • Matthay et al. J Clin Oncol 27, 1007-1013, (2009).
  • Clark, O., Daga, S. & Stoker, A. W. Cancer Lett 328, 44-54, (2013).
  • Karjoo et al. Advanced Drug Delivery Reviews 99, 113-128, (2016).
  • Grosse et al. The FASEB journal 24, 2301-2313, (2010).


Contact
Andrew Stoker; a.stoker@ucl.ac.uk