Nature biomedical engineeringSep 16, 2025

Metabolic imaging tracks the health of engineered immune cells

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗

Updated Jan 20, 2026

Abstract

Optical metabolic imaging (OMI) can accurately predict an oxidative metabolic phenotype associated with higher in vivo potency against neuroblastoma in CAR T cells.

Media composition impacts CAR T cell metabolism and activation strength during manufacturing.
OMI parameters can indicate the cell cycle stage and determine optimal gene transfer conditions.
In a CRISPR-edited anti-GD2 CAR T cell model, OMI measurements correlate with improved metabolic fitness.
Manufacturing conditions optimized through OMI may enhance CAR T cell yield and persistence.

Simplified

Chimeric antigen receptor (CAR) T cell therapy for solid tumours is challenging because of the immunosuppressive tumour microenvironment and a complex manufacturing process. Cellular manufacturing protocols directly impact CAR T cell yield, phenotype and metabolism, which correlates with in vivo potency and persistence. Although metabolic fitness is a critical quality attribute, how T cell metabolic requirements vary throughout the manufacturing process remains unexplored. Here we use optical metabolic imaging (OMI), a non-invasive, label-free method to evaluate single-cell metabolism. Using OMI, we identified the impacts of media composition on CAR T cell metabolism, activation strength and kinetics, and phenotype. We demonstrate that OMI parameters can indicate cell cycle stage and optimal gene transfer conditions for both viral transduction and electroporation-based CRISPR/Cas9. In a CRISPR-edited anti-GD2 CAR T cell model, OMI measurements allow accurate prediction of an oxidative metabolic phenotype that yields higher in vivo potency against neuroblastoma. Our data support OMI as a robust, sensitive analytical tool to optimize manufacturing conditions and monitor cell metabolism for increased CAR T cell yield and metabolic fitness.

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Funding

Competing interests

Competing interests: K. Saha receives honoraria for advisory board membership for Andson Biotech and Notch Therapeutics. C.M.C. receives honoraria for advisory board membership for Bayer, Elephas Biosciences, Nektar Therapeutics, Novartis and WiCell Research Institute. M.C.S., D.L.P., K. Saha and D.C. declare two patents pending based on this work (WO2024229098A3, WO2024229099A1). M.C.S. is an advisory board member for Elephas Biosciences. No other conflicts of interest are reported.

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Metabolic imaging tracks the health of engineered immune cells

Abstract

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Funding

Competing interests

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