Visualizing CAR-T cell Immunotherapy Using 3 Tesla Fluorine-19 MRI

Veronica P Dubois; Olivia C Sehl; Paula J Foster; John A Ronald

doi:10.1007/s11307-021-01672-3

Visualizing CAR-T cell Immunotherapy Using 3 Tesla Fluorine-19 MRI

Mol Imaging Biol. 2022 Apr;24(2):298-308. doi: 10.1007/s11307-021-01672-3. Epub 2021 Nov 16.

Authors

Veronica P Dubois^{1

2}, Olivia C Sehl^{1

2}, Paula J Foster^{1

2}, John A Ronald^{3

4

5}

Affiliations

¹ Robarts Research Institute, London, ON, Canada.
² The Department of Medical Biophysics, Western University, London, ON, Canada.
³ Robarts Research Institute, London, ON, Canada. jronald@robarts.ca.
⁴ The Department of Medical Biophysics, Western University, London, ON, Canada. jronald@robarts.ca.
⁵ Lawson Health Research Institute, London, ON, Canada. jronald@robarts.ca.

Abstract

Purpose: Chimeric antigen receptor (CAR) T cell cancer immunotherapies have shown remarkable results in patients with hematological malignancies and represent the first approved genetically modified cellular therapies. However, not all blood cancer patients respond favorably, serious side effects have been reported, and the treatment of solid tumors has been a challenge. An imaging tool for visualizing the variety of CAR-T cell products in use and being explored could provide important patient-specific data on CAR-T cell location to inform on potential success or failure of treatment as well as off-target toxicities. Fluorine-19 (¹⁹F) magnetic resonance imaging (MRI) allows for the noninvasive detection of ¹⁹F perfluorocarbon (PFC) labeled cells. Our objective was to visualize PFC-labeled (PFC +) CAR-T cells in a mouse model of leukemia using clinical field strength (3 Tesla) ¹⁹F MRI and compare the cytotoxicity of PFC + versus unlabeled CAR-T cells.

Procedures: NSG mice (n = 17) received subcutaneous injections of CD19 + human B cell leukemia cells (NALM6) expressing firefly luciferase in their left hind flank (1 × 10⁶). Twenty-one days later, each mouse received an intratumoral injection of 10 × 10⁶ PFC + CD19-targeted CAR-T cells (n = 6), unlabeled CD19-targeted CAR-T cells (n = 3), PFC + untransduced T cells (n = 5), or an equivalent volume of saline (n = 3). ¹⁹F MRI was performed on mice treated with PFC + CAR-T cells days 1, 3, and 7 post-treatment. Bioluminescence imaging (BLI) was performed on all mice days - 1, 5, 10, and 14 post-treatment to monitor tumor response.

Results: PFC + CAR-T cells were successfully detected in tumors using ¹⁹F MRI on days 1, 3, and 7 post-injection. In vivo BLI data revealed that mice treated with PFC + or PFC - CAR-T cells had significantly lower tumor burden by day 14 compared to untreated mice and mice treated with PFC + untransduced T cells (p < 0.05). Importantly, mice treated with PFC + CAR-T cells showed equivalent cytotoxicity compared to mice receiving PFC - CAR-T cells.

Conclusions: Our studies demonstrate that clinical field strength ¹⁹F MRI can be used to visualize PFC + CAR-T cells for up to 7 days post-intratumoral injection. Importantly, PFC labeling did not significantly affect in vivo CAR-T cell cytotoxicity. These imaging tools may have broad applications for tracking emerging CAR-T cell therapies in preclinical models and may eventually be useful for the detection of CAR-T cells in patients where localized injection of CAR-T cells is being pursued.

Keywords: 19Fluorine magnetic resonance imaging; Bioluminescence imaging; CAR-T cell therapy; Cell tracking; Leukemia.

MeSH terms

Animals
Fluorine*
Humans
Immunotherapy* / methods
Immunotherapy, Adoptive
Magnetic Resonance Imaging
Mice
T-Lymphocytes

Substances

Fluorine