PET-based tracking of CAR T cells and viral gene transfer using a cell surface reporter that binds to lanthanide complexes
Volker Morath, Katja Fritschle, Linda Warmuth, Markus R. Anneser, Sarah Dötsch, Milica Živanić, Luisa Krumwiede, P Bösl, Tarik Bozoglu, Stephanie Robu, Silvana Libertini, Susanne Kossatz, Christian Kupatt, Markus Schwaiger, Katja Steiger, Dirk H. Busch, Arne Skerra, Wolfgang Weber
Abstract
The clinical translation of cell- and gene-based therapies is limited by the lack of non-invasive, quantitative and specific whole-body imaging tools. Here we present a positron emission tomography reporter system based on a membrane-anchored anticalin protein that binds a fluorine-18-labelled lanthanide complex with picomolar affinity via a bio-orthogonal interaction. The reporter was introduced into therapeutic cells, including CAR T cells and adeno-associated virus-transduced cells. In vitro, reporter expression conferred >800-fold higher radioligand binding versus controls. In mice, the radioligand demonstrated rapid renal clearance, showed no off-target accumulation and enabled high-contrast detection of as few as 1,200 CAR T cells in the bone marrow. Longitudinal positron emission tomography imaging over 4 weeks revealed precise tracking of CAR T cell expansion and migration, with signal intensity correlating linearly with flow cytometry data. The system also enabled the quantitative imaging of in vivo gene transfer using an adeno-associated viral vector. This depth-independent whole-body imaging platform offers a powerful tool for monitoring therapeutic cell dynamics and gene delivery in preclinical and potentially clinical settings.