Analysis and stratification of paediatric brain tumours using advanced Raman imaging technology as a

Analysis and stratification of paediatric brain tumours using advanced Raman imaging technology as a potential rapid diagnostic tool.

Background:

Raman spectroscopy (RS) is a non-destructive chemical analysis technique which provides detailed information about chemical structure, phase and molecular interactions. It is based upon the interaction of light with the chemical bonds within a material. Advanced equipment and processing techniques now allow for rapid identification of a sample’s ‘molecular fingerprint’. In recent years, it has been applied to many biodiagnostic applications, including tissue and bacteria identification. More recently this has been used in cancer diagnostics and adult brain tumours. Working in partnership with Renishaw a leader in RS instrument development and Prof Geraint Thomas (UCL Div Bioscience) we have successfully piloted the ability of a prototype RS analyser to distinguish between the 4 most common childhood brain tumour types. There is now an opportunity for a PhD project to move this initial work forward using more advanced RS systems to explore RS as an innovative rapid diagnostic tool for paediatric neuro-oncology, both in the neuropathology lab (RA816 biological analyser) and in the operating theatre (Renishaw Virsa™ in-vivo biological analyser). https://www.renishaw.com/en/raman-spectroscopy--6150

Aims/Objectives:

Phase 1 – Optimise sample preparation and processing to determine sensitivity of the RA816 biological analyser under “NHS laboratory” conditions, i.e. ex-vivo under the microscope to differentiate between paediatric brain tumours subtypes.

•    Generate prediction models for different paediatric brain tumours, comparing against standard of care neuropathology diagnostics for both rapid diagnostic “intra-operative smear” and standard fixed tissue samples”

Phase 2 – Explore feasibility of incorporating the use Virsa™ in-vivo biological analyser within the neurosurgical operating theatre environment, optimising conditions and processing.

•    Determine sensitivity of in-vivo analysis with the Virsa™ in-vivo biological analyser to to differentiate between normal and tumour brain tissue and paediatric brain tumours subtypes.

•    Generate prediction models for different paediatric brain tumours, comparing against standard of care neuropathology diagnostics for both rapid diagnostic “intra-operative smear” and standard fixed tissue samples” and the RA816 biological analyser under “NHS laboratory” conditions.

Phase 3 – Validate in a real-life situation Virsa™ in vivo biological analyser

Methods:

Samples- Initially use archival GOSH neuropathology tumour bank samples previously s fully characterized using NHS “gold standard” integrated molecular neuropathology techniques to optimize sample preparation for fixed tissues for analysis using a Renishaw RA816 benchtop Raman system configured for pathology use. Later moving to fresh biopsy samples.

Data processing- Renishaw have developed dedicated software which can be used to build classification models to analyse tissue using Principal component analysis-fed linear discriminant analysis (PCA-LDA) models. Renishaw and Prof Thomas team will support the student with this data analysis and this will be compared to the NHS “gold standard” integrated molecular neuropathology report (Prof Jacques).

Renishaw Virsa™ in vivo biological analyser- the feasibility of implementing this probe-based system for in-vivo analysis of tissue within the neurosurgical theatre to be used during brain tumour resections/ biopsies will be explored in terms of environmental factors, acquisition times, probe sterilisation and placement and data collection initially in a “non-live” scenario and subsequently after completion of phase 1 and 2 in a real-life operating theatre situation.

References:.

Livermore LJ, et al. Rapid intraoperative molecular genetic classification of gliomas using Raman spectroscopy. Neurooncol Adv. 2019 May-Dec;1(1):vdz008.

Livermore et al. Raman spectroscopy to differentiate between fresh tissue samples of glioma and normal brain: a comparison with 5-ALA-induced fluorescence-guided surgery, J. Neurosurg. 2020

Pekmezci M, et al. Detection of glioma infiltration at the tumor margin using quantitative stimulated Raman scattering histology. Sci Rep. 2021 Jun 9;11(1):12162.

Di L, Eichberg DG, eat al. Stimulated Raman Histology for Rapid Intraoperative Diagnosis of Gliomas. World Neurosurg. 2021 Jun;150:e135-e143.