PMID- 36914208 OWN - NLM STAT- MEDLINE DCOM- 20230315 LR - 20230317 IS - 2051-1426 (Electronic) IS - 2051-1426 (Linking) VI - 11 IP - 3 DP - 2023 Mar TI - Metabolic reprogramming via an engineered PGC-1alpha improves human chimeric antigen receptor T-cell therapy against solid tumors. LID - 10.1136/jitc-2022-006522 [doi] LID - e006522 AB - BACKGROUND: Cellular immunotherapies for cancer represent a means by which a patient's immune system can be augmented with high numbers of tumor-specific T cells. Chimeric antigen receptor (CAR) therapy involves genetic engineering to 'redirect' peripheral T cells to tumor targets, showing remarkable potency in blood cancers. However, due to several resistance mechanisms, CAR-T cell therapies remain ineffective in solid tumors. We and others have shown the tumor microenvironment harbors a distinct metabolic landscape that produces a barrier to immune cell function. Further, altered differentiation of T cells within tumors induces defects in mitochondrial biogenesis, resulting in severe cell-intrinsic metabolic deficiencies. While we and others have shown murine T cell receptor (TCR)-transgenic cells can be improved through enhanced mitochondrial biogenesis, we sought to determine whether human CAR-T cells could be enabled through a metabolic reprogramming approach. MATERIALS AND METHODS: Anti-EGFR CAR-T cells were infused in NSG mice which bore A549 tumors. The tumor infiltrating lymphocytes were analyzed for exhaustion and metabolic deficiencies. Lentiviruses carrying PPAR-gamma coactivator 1alpha (PGC-1alpha), PGC-1alpha(S571A) and NT-PGC-1alpha constructs were used to co-transduce T cells with anti-EGFR CAR lentiviruses. We performed metabolic analysis via flow cytometry and Seahorse analysis in vitro as well as RNA sequencing. Finally, we treated therapeutically A549-carrying NSG mice with either PGC-1alpha or NT-PGC-1alpha anti-EGFR CAR-T cells. We also analyzed the differences in the tumor-infiltrating CAR-T cells when PGC-1alpha is co-expressed. RESULTS: Here, in this study, we show that an inhibition resistant, engineered version of PGC-1alpha, can metabolically reprogram human CAR-T cells. Transcriptomic profiling of PGC-1alpha-transduced CAR-T cells showed this approach effectively induced mitochondrial biogenesis, but also upregulated programs associated with effector functions. Treatment of immunodeficient animals bearing human solid tumors with these cells resulted in substantially improved in vivo efficacy. In contrast, a truncated version of PGC-1alpha, NT-PGC-1alpha, did not improve the in vivo outcomes. CONCLUSIONS: Our data further support a role for metabolic reprogramming in immunomodulatory treatments and highlight the utility of genes like PGC-1alpha as attractive candidates to include in cargo along with chimeric receptors or TCRs for cell therapy of solid tumors. CI - (c) Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ. FAU - Lontos, Konstantinos AU - Lontos K AD - Tumor Microenvironment Center, Department of Immunology, UPMC Hillman Cancer Center and University of Pittsburgh, Pittsburgh, PA, USA. FAU - Wang, Yiyang AU - Wang Y AD - Tumor Microenvironment Center, Department of Immunology, UPMC Hillman Cancer Center and University of Pittsburgh, Pittsburgh, PA, USA. AD - Tsinghua University School of Medicine, Beijing, China. FAU - Joshi, Supriya K AU - Joshi SK AD - Tumor Microenvironment Center, Department of Immunology, UPMC Hillman Cancer Center and University of Pittsburgh, Pittsburgh, PA, USA. FAU - Frisch, Andrew T AU - Frisch AT AD - Tumor Microenvironment Center, Department of Immunology, UPMC Hillman Cancer Center and University of Pittsburgh, Pittsburgh, PA, USA. FAU - Watson, McLane J AU - Watson MJ AD - Tumor Microenvironment Center, Department of Immunology, UPMC Hillman Cancer Center and University of Pittsburgh, Pittsburgh, PA, USA. FAU - Kumar, Alok AU - Kumar A AD - Tumor Microenvironment Center, Department of Immunology, UPMC Hillman Cancer Center and University of Pittsburgh, Pittsburgh, PA, USA. FAU - Menk, Ashley V AU - Menk AV AD - Tumor Microenvironment Center, Department of Immunology, UPMC Hillman Cancer Center and University of Pittsburgh, Pittsburgh, PA, USA. FAU - Wang, Yupeng AU - Wang Y AD - Tumor Microenvironment Center, Department of Immunology, UPMC Hillman Cancer Center and University of Pittsburgh, Pittsburgh, PA, USA. AD - Tsinghua University School of Medicine, Beijing, China. FAU - Cumberland, Rachel AU - Cumberland R AD - Tumor Microenvironment Center, Department of Immunology, UPMC Hillman Cancer Center and University of Pittsburgh, Pittsburgh, PA, USA. FAU - Lohmueller, Jason AU - Lohmueller J AUID- ORCID: 0000-0001-6089-9992 AD - Department of Surgery, UPMC Hillman Cancer Center, University of Pittsburgh, Pittsburgh, PA, USA. FAU - Carrizosa, Esteban AU - Carrizosa E AD - 2seventybio, Boston, Massachusetts, USA. FAU - Boyerinas, Benjamin AU - Boyerinas B AD - 2seventybio, Boston, Massachusetts, USA. FAU - Delgoffe, Greg M AU - Delgoffe GM AUID- ORCID: 0000-0002-2957-8135 AD - Tumor Microenvironment Center, Department of Immunology, UPMC Hillman Cancer Center and University of Pittsburgh, Pittsburgh, PA, USA delgoffeg@upmc.edu. LA - eng GR - P30 CA047904/CA/NCI NIH HHS/United States GR - TL1 TR001858/TR/NCATS NIH HHS/United States GR - T32 CA082084/CA/NCI NIH HHS/United States GR - F31 AI149971/AI/NIAID NIH HHS/United States GR - DP2 AI136598/AI/NIAID NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't PL - England TA - J Immunother Cancer JT - Journal for immunotherapy of cancer JID - 101620585 RN - 0 (Receptors, Chimeric Antigen) RN - 0 (Receptors, Antigen, T-Cell) SB - IM MH - Humans MH - Animals MH - Mice MH - *Receptors, Chimeric Antigen MH - Immunotherapy, Adoptive/methods MH - *Neoplasms MH - Receptors, Antigen, T-Cell MH - T-Lymphocytes MH - Tumor Microenvironment PMC - PMC10016249 OTO - NOTNLM OT - Cell Engineering OT - Immunotherapy, Adoptive OT - Receptors, Chimeric Antigen OT - Translational Medical Research COIS- Competing interests: GMD has filed patent applications around the use of PGC-1alpha as a metabolic reprogramming agent in cellular therapies. BB was an employee of 2seventy bio during the period of the research and owns stock of the same company. EC was an employee of 2seventy bio during the period of the research. EDAT- 2023/03/14 06:00 MHDA- 2023/03/16 06:00 PMCR- 2023/03/13 CRDT- 2023/03/13 20:52 PHST- 2023/02/08 00:00 [accepted] PHST- 2023/03/13 20:52 [entrez] PHST- 2023/03/14 06:00 [pubmed] PHST- 2023/03/16 06:00 [medline] PHST- 2023/03/13 00:00 [pmc-release] AID - jitc-2022-006522 [pii] AID - 10.1136/jitc-2022-006522 [doi] PST - ppublish SO - J Immunother Cancer. 2023 Mar;11(3):e006522. doi: 10.1136/jitc-2022-006522.