Fluorescence-guided surgical sampling of glioblastoma identifies phenotypically distinct tumour-initiating cell populations in the tumour mass and margin

• Published in: British journal of cancer 2012-07, Vol.107 (3), p.462-468
• Main Authors: Piccirillo, S G M, Dietz, S, Madhu, B, Griffiths, J, Price, S J, Collins, V P, Watts, C
• Format: Article
• Language: English
• Subjects: 631/1647/245/2225; 692/699/67/1922; 692/700/565/545/546; Adult; Aged; Aminolevulinic Acid - metabolism; Animals; Biological and medical sciences; Biomedical and Life Sciences; Biomedicine; Brain cancer; Brain Neoplasms - metabolism; Brain Neoplasms - pathology; Brain Neoplasms - surgery; Brain research; Cancer Research; Cell Transformation, Neoplastic - metabolism; Cell Transformation, Neoplastic - pathology; Drug Resistance; Epidemiology; Female; Fluorescence; Glioblastoma - metabolism; Glioblastoma - pathology; Glioblastoma - surgery; Humans; Magnetic Resonance Imaging - methods; Male; Medical research; Medical sciences; Metabolism; Metabolome; Mice; Mice, Inbred NOD; Mice, SCID; Middle Aged; Molecular Medicine; Neurology; Neurosciences; Oncology; Phenotype; R&D; Research & development; Surgery; Translational Therapeutics; Tumor Cells, Cultured; Tumors; Tumors of the nervous system. Phacomatoses
• ISSN: 0007-0920; 1532-1827
• DOI: 10.1038/bjc.2012.271

Background: Acquiring clinically annotated, spatially stratified tissue samples from human glioblastoma (GBM) is compromised by haemorrhage, brain shift and subjective identification of ‘normal’ brain. We tested the use of 5-aminolevulinic acid (5-ALA) fluorescence to objective tissue sampling and to derive tumour-initiating cells (TICs) from mass and margin. Methods: The 5-ALA was administered to 30 GBM patients. Samples were taken from the non-fluorescent necrotic core, fluorescent tumour mass and non-fluorescent margin. We compared the efficiency of isolating TICs from these areas in 5-ALA versus control patients. HRMAS 1 H NMR was used to reveal metabolic alterations due to 5-ALA. We then characterised TICs for self-renewal in vitro and tumorigenicity in vivo . Results: The derivation of TICs was not compromised by 5-ALA and the metabolic profile was similar between tumours from 5-ALA patients and controls. The TICs from the fluorescent mass were self-renewing in vitro and tumour-forming in vivo , whereas TICs from non-fluorescent margin did not self-renew in vitro but did form tumours in vivo . Conclusion: Our data show that 5-ALA does not compromise the derivation of TICs. It also reveals that the margin contains TICs, which are phenotypically different from those isolated from the corresponding mass.