Antibody-target binding quantification in living tumors using macroscopy fluorescence lifetime Forster resonance energy transfer imaging (MFLI FRET)

Nanxue Yuan; Vikas Pandey; Amit Verma; John C. Williams; Xavier Intes; Margarida Barroso

doi:10.1117/12.3003282

12 March 2024 Antibody-target binding quantification in living tumors using macroscopy fluorescence lifetime Forster resonance energy transfer imaging (MFLI FRET)

Nanxue Yuan, Vikas Pandey, Amit Verma, John C. Williams, Xavier Intes, Margarida Barroso

Author Affiliations +

Proceedings Volume 12821, Visualizing and Quantifying Drug Distribution in Tissue VIII; 1282105 (2024) https://doi.org/10.1117/12.3003282
Event: SPIE BiOS, 2024, San Francisco, California, United States

Abstract

Emergent breast tumor resistance and tumor microenvironment (TME) heterogeneity can lead to decreased drug delivery efficacy, resulting in therapeutic failure. Preclinical molecular imaging is a crucial tool in the advancement of targeted therapeutics for supporting the development of new drugs but also to elucidate factors hampering optimal drug delivery. However, noninvasive tumor imaging modalities that can quantify drug-target engagement, which is critical for therapeutic actuation, are still lacking. We have demonstrated the utility of macroscopic fluorescence lifetime Forster’s Resonance Energy Transfer (MFLI FRET)-based optical imaging to measure labeled trastuzumab (TZM)-human epidermal growth factor receptor (HER2) binding in human HER2+ cell lines and breast tumor xenograft mice models. We have established a clinically relevant TZM antibody drug containing the Meditope (MDT) peptide conjugated to near-infrared (NIR) dyelabeled FRET pairs, that retain full HER2 binding capability. Herein, we demonstrate FRET measurements using MFLI in vivo imaging platform to measure the ability of MDT-TZM to bind HER2 in living breast tumor xenografts. HER2+ AU565 breast tumor xenografts bearing nude mice were injected retro-orbitally with TZM (NHS-conjugated) or MDTTZM labeled with AlexaFluor700 (donor) and AlexaFluor750 (acceptor) and MFLIFRET imaging was performed 24 h and 48 h post-injection. Preliminary data suggest that MDT-TZM shows higher uniform and consistent FRET signal compared to TZM, suggesting increased efficacy of TZM-MDT-HER2 binding. Also staggered injections of donor and acceptor MDT-TZM may be optimal for quantifying MDT-TZM using MFLIFRET compared to single injections.

Conference Presentation

(2024) Published by SPIE. Downloading of the abstract is permitted for personal use only.

Citation Download Citation

Nanxue Yuan, Vikas Pandey, Amit Verma, John C. Williams, Xavier Intes, and Margarida Barroso "Antibody-target binding quantification in living tumors using macroscopy fluorescence lifetime Forster resonance energy transfer imaging (MFLI FRET)", Proc. SPIE 12821, Visualizing and Quantifying Drug Distribution in Tissue VIII, 1282105 (12 March 2024); https://doi.org/10.1117/12.3003282

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