Journal Article

Feasibility of real-time in vivo 89Zr-DFO-labeled CAR T-cell trafficking using PET imaging

Sheen, Joong Hyuk, Kim, Hyori, Chung, Junho, Ju, Ji-Min, Lee, Sang Ju, Ryu, Jin-Sook, Chung, Jin Hwa, Choi, Kyungho, Cho, Eun Hye, Soh, Hyunsu, Lee, Sang-Jin, Oh, Seung Jun, Kim, Seog-Young, Lee, Suk Hyun

2020

Overview

Chimeric antigen receptor (CAR) T-cells have been recently developed and are producing impressive outcomes in patients with hematologic malignancies. However, there is no standardized method for cell trafficking and in vivo CAR T-cell monitoring. We assessed the feasibility of real-time in vivo 89Zr-p-Isothiocyanatobenzyl-desferrioxamine (Df-Bz-NCS, DFO) labeled CAR T-cell trafficking using positron emission tomography (PET). The 89Zr-DFO radiolabeling efficiency of Jurkat/CAR and human peripheral blood mononuclear cells (hPBMC)/CAR T-cells was 70%-79%, and cell radiolabeling activity was 98.1-103.6 kBq/106 cells. Cell viability after radiolabeling was >95%. Cell proliferation was not significantly different during the early period after radiolabeling, compared with unlabeled cells; however, the proliferative capacity decreased over time (day 7 after labeling). IL-2 or IFN-γ secretion was not significantly different between unlabeled and labeled CAR T-cells. PET/magnetic resonance imaging in the xenograft model showed that most of the 89Zr-DFO-labeled Jurkat/CAR T-cells were distributed in the lung (24.4% ± 3.4%ID) and liver (22.9% ± 5.6%ID) by one hour after injection. The cells gradually migrated from the lung to the liver and spleen by day 1, and remained stable in these sites until day 7 (on day 7: lung 3.9% ± 0.3%ID, liver 36.4% ± 2.7%ID, spleen 1.4% ± 0.3%ID). No significant accumulation of labeled cells was identified in tumors. A similar pattern was observed in ex vivo biodistributions on day 7 (lung 3.0% ± 1.0%ID, liver 19.8% ± 2.2%ID, spleen 2.3% ± 1.7%ID). 89Zr-DFO-labeled hPBMC/CAR T-cells showed a similar distribution, compared with Jurkat/CAR T-cells, on serial PET images. CAR T cell distribution was cross-confirmed by flow cytometry, Alu polymerase chain reaction, and immunohistochemistry. Real-time in vivo cell trafficking is feasible using PET imaging of 89Zr-DFO-labeled CAR T-cells. This can be used to investigate cellular kinetics, initial in vivo biodistribution, and safety profiles in future CAR T-cell development.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Antigens / Biology and Life Sciences / Blood cancer / Cancer therapies / Cell adhesion & migration / Cell proliferation / Cell Proliferation - drug effects / Cell Survival - drug effects / Cell viability / Chimeric antigen receptors / Deferoxamine / Deferoxamine - analogs & derivatives / Deferoxamine - pharmacology / Experiments / Feasibility studies / Flow cytometry / Genetic engineering / Hematologic Neoplasms - drug therapy / Hematologic Neoplasms - pathology / Humans / Immunoconjugates - pharmacology / Immunohistochemistry / Immunotherapy / In vivo methods and tests / Interleukin 2 / Isothiocyanates - pharmacology / Isotope Labeling / Jurkat Cells / Leukocytes (mononuclear) / Leukocytes, Mononuclear - chemistry / Leukocytes, Mononuclear - drug effects / Life sciences / Liver / Lungs / Lymphocytes / Lymphocytes T / Magnetic resonance / Magnetic resonance imaging / Medical imaging / Medical schools / Medicine and Health Sciences / Methods / Nuclear medicine / Peripheral blood mononuclear cells / Physical Sciences / Polymerase chain reaction / Positron emission / Positron emission tomography / Radioisotopes / Radioisotopes - chemistry / Radioisotopes - pharmacology / Radiolabelling / Reaction kinetics / Real time / Receptors, Antigen, T-Cell - chemistry / Receptors, Antigen, T-Cell - isolation & purification / Receptors, Antigen, T-Cell - therapeutic use / Receptors, Chimeric Antigen - chemistry / Receptors, Chimeric Antigen - isolation & purification / Receptors, Chimeric Antigen - therapeutic use / Research and Analysis Methods / Spleen / Studies / T-Lymphocytes - chemistry / T-Lymphocytes - immunology / Tissue Distribution / Tomography / Trafficking / Tumors / Viability / Xenografts / Xenotransplantation / Zirconium - pharmacology / Zirconium isotopes / γ-Interferon