Metabolic profiling of CAR T-cells in patients reveals a shift toward amino acid-supported OXPHOS and informs transporter engineering
Year: 2026
Type: preprint
Abstract: CAR T-cell efficacy requires post-infusion expansion and persistence, yet metabolic programs supporting T-cells in patients remain poorly defined. Here, we developed a high-throughput single-cell immunometabolic profiling pipeline and applied it across pediatric leukemia trials, revealing a conserved post-infusion CAR T-cell shift from glycolysis toward amino acid-driven oxidative phosphorylation (OXPHOS). Within this remodeling, OXPHOS-dependent stem-like CAR T-cell subsets were enriched in patients achieving complete remission. Longitudinal plasma metabolomics revealed cytokine release syndrome-associated depletion of multiple amino acids, including glutamine and arginine, during CAR T-cell expansion, creating a nutrient-restricted environment. Analysis of public CAR T-cell datasets showed that responders upregulated amino-acid solute carrier transporters, whereas disrupting uptake impaired translation, OXPHOS, stemness, and cytotoxicity. Guided by these findings, we screened amino-acid transporters for CAR T-cell engineering. SLC1A5-, SLC7A1-, and SLC38A9-armored CAR T-cells emerged as the most promising, with enhanced oxidative capacity and anti-leukemic efficacy, establishing amino acid transport as a targetable metabolic checkpoint.
Source: Research Square
Authors: Naomi Taylor, Josquin Moraly, Taisuke Kondo, King Chan, Sooraj Achar, Makoto Ando, Marie Pouzolles, Bonnie Yates, Alexandra Dreyzin, Mehdi Benzaoui, Alka Dwivedi, Saliha Majdoul, Justin Mirazee, Jaehyun Suh, Angela Su, Cedric Mongellaz, Hannah Dada, Swapna Vidhur Dautalabad, Krithikha Bhuvaneshwar, Mina O Seedhom, Valerie Zimmermann, Sandrina Kinet, Daniel Crooks, Hannah Song, Jonathan Yewdell, Christopher Chien, Olivier Hermine, Valerie Dardhalon, Steven Highfill, Thorkell Andresson, Grégoire Altan-Bonnet, Nirali Shah
Institutions: National Institutes of Health, Keio University, Frederick National Laboratory for Cancer Research, Texas Children's Hospital, University of Colorado Anschutz, Weill Cornell Medicine, AstraZeneca (United Kingdom), University of Oxford, Georgetown University, University of Massachusetts Chan Medical School, Institut de Génétique Moléculaire de Montpellier, Centre National de la Recherche Scientifique, National Cancer Institute, Université Paris Descartes
Language: English
Topic: CAR-T cell therapy research
Subfield: Oncology
Field: Medicine
Domain: Health Sciences
Open Access status: green
Funder: National Cancer Institute
OpenAlex ID: W7213330534