UCSF

Purpose

To assess the cell-specific, intracellular partial pressure of oxygen (Po₂) dynamics of both tumor and chimeric antigen receptor (CAR) T cells in a murine immunotherapy model.

Materials and methods

Human glioblastoma cells or human T cells were intracellularly labeled with perfluorocarbon nanoemulsion droplet sensors prior to in vivo injection in severe combined immunodeficient mice to measure Po₂ in the two cell types in response to treatment. Two main sets of experiments were performed: (a) mice were injected in the flank with perfluorocarbon-labeled human glioblastoma cells and were then inoculated with either CAR T cells or untransduced T cells or were untreated 5 days after tumor inoculation; and (b) mice with unlabeled glioblastoma tumors were inoculated with perfluorocarbon-labeled CAR T cells or untransduced T cells 5 days after tumor inoculation. Longitudinal fluorine 19 (¹⁹F) spin-lattice relaxation time measurements of the tumor mass were used to ascertain absolute Po₂ in vivo. Results were analyzed for significance using an analysis of variance, a linear mixed-effect model, and a Pearson correlation coefficient test, as appropriate.

Results

The intracellular tumor cell Po₂ temporal dynamics exhibited delayed, transient hyperoxia at 3 days after infusion of CAR T cells, commensurate with significant tumor cell killing and CAR T-cell infiltration, as observed by bioluminescence imaging and histologic findings. Conversely, no significant changes were detected in CAR or untransduced T-cell intracellular Po₂ over time in tumor using these same methods. Moreover, it was observed that the total ¹⁹F tumor cell signal quenches with treatment, consistent with rapid tissue clearance of probe from apoptotic tumor cells.

Conclusion

Cell-specific Po₂ measurements using perfluorocarbon probes can provide insights into effector cell function and tumor response in cellular immunotherapeutic cancer models.Keywords: Animal Studies, MR-Imaging, MR-Spectroscopy, Molecular Imaging-Cancer, Molecular Imaging-Immunotherapy Supplemental material is available for this article. © RSNA, 2021See also commentary by Bulte in this issue.