Session: 702. CAR-T Cell Therapies: Basic and Translational: Poster III
Hematology Disease Topics & Pathways:
Fundamental Science, Research, Translational Research, Immunology, Metabolism, Biological Processes, Molecular biology
Flow cytometry, mitochondrial stress tests and metabolomics were used to analyze healthy and CLL patients' T cells at baseline and post-T-cell receptor (TCR) activation. For murine experiments, splenocytes obtained from transgenic Eμ-TCL1 mice were adoptively transferred (AT) into wild-type C57BL/6 mice, and splenic T cells were studied at various disease stages. Regarding ex vivo reprogramming experiments, human or mouse T-cell cultures were treated with the PI3Kδ inhibitor idelalisib. Generation of CAR T cells was done using T cells from AT Eμ-TCL1 mice, stimulated ex vivo with or without idelalisib, and virally transduced with a GFP-tagged m1928z CAR construct. Leukemic mice infused with these CAR T cells were monitored over time.
Mitochondrial depolarization, indicated by a reduced mitochondrial membrane potential relative to mass, was observed in human and murine CLL T cells. T cells from AT Eμ-TCL1 mice demonstrated disturbed mitochondrial function alongside a shift toward terminally differentiated exhausted-like cells during CLL progression. Furthermore, extracellular flux analysis following TCR stimulation indicated diminished mitochondrial spare respiratory capacity in CLL T cells, consistent in both human and mouse models. Metabolomics and 13C fractional labeling demonstrated decreased mitochondrial fueling and lower levels of mitochondrial metabolites in CLL T cells. These observations of metabolic disturbance aligned with an exhausted-like T-cell differentiation phenotype controlled at the epigenetic level. An increase in PI3K/Akt signaling was identified in T cells from AT Eμ-TCL1 mice. Ex vivo reprogramming of human and murine CLL T cells with the PI3Kδ inhibitor idelalisib resulted in metabolically reprogrammed T cells with improved memory formation and mitochondrial activity. Adding idelalisib during the generation of murine CD19 CAR T cells significantly enhanced the in vivo persistence of infused cells in the immunocompetent Eμ-TCL1 mice and led to long-term leukemia-free remissions, underscoring the clinical relevance of these findings.
Overall, we found that the buildup of dysfunctional, depolarized mitochondria is a marker of T-cell metabolic abnormalities associated with terminal differentiation in CLL progression. PI3K inhibition is a potential strategy to improve CLL T-cell metabolic plasticity and enhance the efficacy and persistence of autologous CAR T-cell infusion products.
WG, NG, and HSM share the first co-authorship.
JPI and APK share senior authorship.
Disclosures: Kater: Roche/Genentech: Membership on an entity's Board of Directors or advisory committees, Research Funding; BMS: Membership on an entity's Board of Directors or advisory committees, Research Funding; LAVA: Membership on an entity's Board of Directors or advisory committees, Other: Patents planned, issued or pending; Steering Committee; Janssen: Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Patents planned, issued or pending; Steering Committee, Research Funding; AbbVie: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; AstraZeneca: Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Steering Committee, Research Funding. Pinilla-Ibarz: Bristol Meyers Squibb: Consultancy, Speakers Bureau; Novartis: Honoraria; AbbVie: Consultancy, Speakers Bureau; AstraZeneca: Consultancy, Speakers Bureau; Eli Lily: Consultancy, Speakers Bureau; Secura Bio: Consultancy, Speakers Bureau; Sanofi: Consultancy, Speakers Bureau; Pfizer: Consultancy; Beigene: Consultancy, Speakers Bureau; Janssen: Consultancy, Speakers Bureau; Takeda: Consultancy, Speakers Bureau.
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