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posted on 2023-06-15, 08:20 authored by Laura Privitera, Dale J. Waterhouse, Alessandra Preziosi, Irene Paraboschi, Olumide Ogunlade, Chiara Da Pieve, Marta Barisa, Olumide Ogunbiyi, Gregory Weitsman, J. Ciaran Hutchinson, Kate Cross, Lorenzo Biassoni, Danail Stoyanov, Neil Sebire, Paul Beard, Paolo De Coppi, Gabriela Kramer-Marek, John Anderson, Stefano Giuliani In vitro validation of anti–GD2-IR800 and anti–GD2-IR12. A, Binding specificity of anti–GD2-IR800 and anti–GD2-IR12 conjugates assessed by flow cytometry in three GD2-positive cell lines (KELLY, LAN-1, and SUPT1-GD2) and in the negative control (SUPT1-WT). B, Both conjugates show higher SBR (median stained fluorescence intensity/median unstained fluorescence intensity) for GD2-positive cell lines than for GD2-negative cell lines. Box represents interquartile range. Bars represent 5% to 95% range. Number of cells = 44,810–70,831. C, Fluorescence and brightfield images of the adherent cells 2 hours after staining with 100 nmol/L of either anti–GD2-IR800 or anti–GD2-IR12 (red). The nuclear counterstain was DAPI (blue). D, Representative section from the Z-stack acquisition of LAN-1 spheroids 24 hours after staining with 100 nmol/L of either anti–GD2-IR800 or anti–GD2-IR12 (red). The nuclear counterstain was DAPI (blue).
Funding
NIHR Great Ormond Street Hospital Biomedical Research Centre (BRC)
research into childhood cancer
GOSHCC Cancer infrastructure award
Royal Academy of Engineering chair in emerging Technologies Scheme
Medical Research Council (MRC)
Wellcome Trust (WT)
University College London Wellcome/EPSRC Centre for Interventional and Surgical Sciences
Engineering and Physical Sciences Research Council (EPSRC)
Cancer Research UK (CRUK)
European Research Council (ERC)
History
ARTICLE ABSTRACT
Fluorescence-guided surgery is set to play a pivotal role in the intraoperative management of pediatric tumors. Shortwave infrared imaging (SWIR) has advantages over conventional near-infrared I (NIR-I) imaging with reduced tissue scattering and autofluorescence. Here, two NIR-I dyes (IRDye800CW and IR12), with long tails emitting in the SWIR range, were conjugated with a clinical-grade anti-GD2 monoclonal antibody (dinutuximab-beta) to compare NIR-I and SWIR imaging for neuroblastoma surgery. A first-of-its-kind multispectral NIR-I/SWIR fluorescence imaging device was constructed to allow an objective comparison between the two imaging windows. Conjugates were first characterized in vitro. Tissue-mimicking phantoms, imaging specimens of known geometric and material composition, were used to assess the sensitivity and depth penetration of the NIR-I/SWIR device, showing a minimum detectable volume of ∼0.9 mm3 and depth penetration up to 3 mm. In vivo, fluorescence imaging using the NIR-I/SWIR device showed a high tumor-to-background ratio (TBR) for both dyes, with anti–GD2-IR800 being significantly brighter than anti–GD2-IR12. Crucially, the system enabled higher TBR at SWIR wavelengths than at NIR-I wavelengths, verifying SWIR imaging enables high-contrast delineation of tumor margins. This work demonstrates that by combining the high specificity of anti-GD2 antibodies with the availability and translatability of existing NIR-I dyes, along with the advantages of SWIR in terms of depth and tumor signal-to-background ratio, GD2-targeted NIR-I/SWIR-guided surgery could improve the treatment of patients with neuroblastoma, warranting investigation in future clinical trials.
Multispectral near-infrared I/shortwave infrared fluorescence imaging is a versatile system enabling high tumor-to-background signal for safer and more complete resection of pediatric tumors during surgery.
