Imaging CAR T Cell Trafficking with eDHFR as a PET Reporter Gene

Sellmyer, Mark A.; Richman, Sarah A.; Lohith, Katheryn; Hou, Catherine; Weng, Chi-Chang; Mach, Robert H.; O’Connor, Roddy S.; Milone, Michael C.; Farwell, Michael D.

doi:10.1016/j.ymthe.2019.10.007

Cited by 74 publications

(52 citation statements)

References 36 publications

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“…PET scans show colocalization between anti-GD2 CAR-T cells and GD2 + tumor, which was confirmed with bioluminescence. Finally, they calculated that their method can detect ~11,000 CD8 + CAR-T cells per mm 3 ( 74 ). Park et al used human somatostatin receptor 2 (SSTR2), which ensures intracellular accumulation of 18 F-NOTA-Octreotide (NOTAOCT), to track CAR-T cells of differing affinities to ICAM-1.…”

Section: Car Detection At the Organismal Levelmentioning

confidence: 99%

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The Chimeric Antigen Receptor Detection Toolkit

Huang

2020

Front. Immunol.

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Chimeric antigen receptor-T (CAR-T) cell therapy is a promising frontier of immunoengineering and cancer immunotherapy. Methods that detect, quantify, track, and visualize the CAR, have catalyzed the rapid advancement of CART cell therapy from preclinical models to clinical adoption. For instance, CAR-staining/labeling agents have enabled flow cytometry analysis, imaging applications, cell sorting, and high-dimensional clinical profiling. Molecular assays, such as quantitative polymerase chain reaction, integration site analysis, and RNA-sequencing, have characterized CAR transduction, expression, and in vivo CART cell expansion kinetics. In vitro visualization methods, including confocal and total internal reflection fluorescence microscopy, have captured the molecular details underlying CAR immunological synapse formation, signaling, and cytotoxicity. In vivo tracking methods, including two-photon microscopy, bioluminescence imaging, and positron emission tomography scanning, have monitored CART cell biodistribution across blood, tissue, and tumor. Here, we review the plethora of CAR detection methods, which can operate at the genomic, transcriptomic, proteomic, and organismal levels. For each method, we discuss: (1) what it measures; (2) how it works; (3) its scientific and clinical importance; (4) relevant examples of its use; (5) specific protocols for CAR detection; and (6) its strengths and weaknesses. Finally, we consider current scientific and clinical needs in order to provide future perspectives for improved CAR detection.

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Section: Car Detection At the Organismal Levelmentioning

confidence: 99%

“…(74).Park et al used human somatostatin receptor 2 (SSTR2), which ensures intracellular accumulation of 18 F-NOTA-Octreotide (NOTAOCT), to track CAR-T cells of differing affinities to ICAM-1. PET scans captured CAR-T cell expansion and contraction kinetics (75).…”

mentioning

confidence: 99%

The Chimeric Antigen Receptor Detection Toolkit

Huang

2020

Front. Immunol.

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“…In vivo monitoring of CAR T cell therapy using molecular imaging is an area of intensive investigation. Several groups, including ours, have explored the utility of reporter genes (11)(12)(13)(14)(15) or ex vivo radiolabeling approaches (16)(17)(18) to allow tracking of CAR T cell therapies with PET. The use of immunoPET has several advantages over these approaches.…”

Section: Discussionmentioning

confidence: 99%

“…To date, several studies using positron emission tomography (PET) imaging have been conducted, due to the high sensitivity and quantitative capabilities of this clinically relevant imaging modality. The majority of the preclinical and clinical pilot studies published so far involves the use of reporter genes inserted into the CAR construct for the detection of CAR T cells (11)(12)(13)(14)(15).…”

Section: Introductionmentioning

confidence: 99%

Molecular Imaging of Chimeric Antigen Receptor T Cells by ICOS-ImmunoPET

Simonetta

Alam

Lohmeyer

et al. 2021

Clinical Cancer Research

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In vivo Imaging of Activated Chimeric Antigen Receptor T Cells. Conflict-of-interest disclosures SSG is the founder and equity holder of CellSight Inc. that develops and translates strategies for imaging cell trafficking/transplantation. CLM holds several patent applications in the area of CAR T cell immunotherapy, is a founder of, holds equity in, and receives consulting fees from Lyell Immunopharma, has received consulting fees from NeoImmune Tech, Nektar Therapeutics and Apricity Health and royalties from Juno Therapeutics for the CD22-CAR. All other authors have declared that no conflict of interest exists. Significance ICOS-immunoPET enables in vivo imaging of activated CAR T cells at the tumor site without the need for incorporation of reporter transgenes or ex vivo biolabeling. Research.

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“…Escherichia coli dihydrofolate reductase enzyme (eDHFR) has been used as a reporter for imaging of GD2-CAR T cells with [ 18 F]-labeled trimethoprim ([ 18 F]-TMP) ( 16 ). PET/CT on days 7 and 13 post CAR T cell administration showed localization of the engineered cells initially in the spleen followed by accumulation at the GD2+ tumors.…”

Section: Reporter-based Imaging Of Car T Cellsmentioning

confidence: 99%

CAR Chase: Where Do Engineered Cells Go in Humans?

et al. 2020

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Chimeric antigen receptor (CAR) – and T-cell receptor (TCR) – modified T-cells are rapidly emerging as a viable treatment option for cancer patients. While initial clinical trials for these CAR T cells showed response rates of over 90% in some cases, retrospective studies have revealed a wide variability in patient responses as well as a significant proportion of patients relapsing after an initial response. In addition, patients often have severe adverse reactions to this therapy (e.g., cytokine release and neurologic syndromes). As a result, much research is still needed to be able to predict both therapeutic outcomes and possible toxicities. Furthermore, little success has been seen in treating solid tumors with engineered T cells and uncovering modes of failure is a topic of much research. Finally, little is known about the T cells’ pharmacokinetics after infusion into the patient, as standard methods of tracking the cells analyze peripheral blood and tumor biopsies – both of which lack spatiotemporal information. Herein, we propose that reporter gene-based imaging of engineered T cells in humans would be tremendously valuable in elucidating the fate of the transplanted T cells and would greatly facilitate clinical translation of new CAR and TCR technologies. Currently, there are no FDA-approved reporter genes and few methods have advanced to human studies. Herein, we outline current reporter gene approaches to track engineered cells in vivo , analyze why current reporter genes have not progressed into the clinic, and propose “rules” for designing a widely applicable reporter gene for use in humans.

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Imaging CAR T Cell Trafficking with eDHFR as a PET Reporter Gene

Cited by 74 publications

References 36 publications

The Chimeric Antigen Receptor Detection Toolkit

The Chimeric Antigen Receptor Detection Toolkit

Molecular Imaging of Chimeric Antigen Receptor T Cells by ICOS-ImmunoPET

CAR Chase: Where Do Engineered Cells Go in Humans?

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