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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 Imaging of stained cell pellets beneath tissue-mimicking material using multispectral NIR-I/SWIR fluorescence imaging. A, Microcentrifuges tubes containing pellets of 2 × 106 GD2-positive cells (LAN-1) and GD2-negative cells (SUPT1-WT) stained with 100 nmol/L of anti–GD2-IR800 and covered with a 2% emulsion intralipid. The pellet is located at the tip of the tube. The fluid level is marked in red on the Petri dish. B and C, Multispectral NIR-I/SWIR fluorescence images were captured. Images show the anti–GD2-IR800–stained GD2-positive cells (LAN-1; B) and the anti–GD2-IR800–stained GD2-negative cells (SUPT1-WT; C). White dotted lines show the approximate location of the microcentrifuge tube. D, Line profiles across the fluorescence images were fitted with a Gaussian plus linear background to extract a height and FWHM. Fits shown as a black line. E–G, Bar graphs of the height (E and F) and the FWHM (G and H) versus depth for each wavelength band for GD2-positive (LAN-1) cells stained with anti–GD2-IR800 (E and G) and anti–GD2-IR12 (F and H).
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.
