Metabolic and epigenetic orchestration of (CAR) T cell fate and function

Akbari, Behnia; Hosseini, Zahra; Shahabinejad, Pardis; Ghassemi, Saba; O’Connor, Roddy S.

doi:10.1016/j.canlet.2022.215948

Cited by 10 publications

(6 citation statements)

References 175 publications

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“…Previous studies have shown that PBMCs and T cells from cancer patients have inherent metabolic and epigenetic dysregulations [ 15 – 17 ]. To investigate the potential differences in immune inhibitory receptor expression, we measured the baseline expression of PD1, TIM3, and A2aR in patient-derived PBMCs and T cells and compared them with healthy controls (Gating strategy is shown in Figure S1 ).…”

Section: Resultsmentioning

confidence: 99%

The inhibitory receptors PD1, Tim3, and A2aR are highly expressed during mesoCAR T cell manufacturing in advanced human epithelial ovarian cancer

et al. 2023

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Background Chemotherapy and surgery have been the mainstays of epithelial ovarian cancer (EOC) treatment so far. Cellular immunotherapies such as CAR T cell therapy have recently given hope of a cure for solid tumors like EOC. However, extrinsic factors associated with the CAR T cell manufacturing process and/or intrinsic dysregulation of patient-derived T cells, which could be associated with cancer itself, cancer stage, and treatment regimen, may hamper the efficacy of CAR T cell therapy and promote their exhaustion or dysfunction. Methods To investigate the association of these factors with CAR T cell exhaustion, the frequency of T and CAR T cells expressing three immune inhibitory receptors (i.e., TIM3, PD1, A2aR) generated from T cells of EOC patients and healthy controls was measured during each stage of CAR T cell production. Results Our findings revealed that primary T cells from EOC patients show significantly elevated expression of immune inhibitory receptors, and this increase was more prominent in patients undergoing chemotherapy and those with advanced cancer. In addition, the CAR T cell manufacturing process itself was found to upregulate the expression of these inhibitory receptors and more importantly increase the population of exhausted mesoCAR T cells. Conclusions Our observations suggest that intrinsic characteristics of patient-derived T cells and extrinsic factors in CAR T cell production protocols should be considered and properly counteracted during CAR T cell manufacturing process. In addition, mitigating the signaling of immune inhibitory receptors through pharmacological/genetic perturbation during CAR T cell manufacturing might profoundly improve CAR T cells function and their antitumor activity in EOC and other solid tumors.

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Section: Resultsmentioning

confidence: 99%

The inhibitory receptors PD1, Tim3, and A2aR are highly expressed during mesoCAR T cell manufacturing in advanced human epithelial ovarian cancer

et al. 2023

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“…Machine learning models based on label-free OMI measurements accurately identified gene transfer conditions that later resulted in high CAR transgene incorporation efficiency, indicating that OMI could inform the timing of genome editing to improve CAR yield. Ex vivo metabolic perturbation during manufacturing, such as transient glucose or glutamine restriction, have been shown to reprogram CAR T cells epigenetically and prime them for the metabolically stressed, immunosuppressive in vivo tumor microenvironment (58,59). Here, we found that CAR T cells undergoing media switch from glucose low glutamine low TexMACS preelectroporation to glucose high glutamine high ImmunoCult XF post-electroporation exhibited distinct OMI profiles, with high oxidative phosphorylation and low glycolytic activity, together with improved in vivo potency.…”

Section: Discussionmentioning

confidence: 99%

Label free metabolic imaging to enhance the efficacy of chimeric antigen receptor T cell therapy

Skala,

Pham,

Cappabianca

et al. 2024

Preprint

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Chimeric antigen receptor (CAR) T cell therapy for solid tumors is challenging not only because of the immunosuppressive tumor microenvironment, but also because of a complex manufacturing process that is difficult to monitor. Manufacturing directly impacts CAR T cell yield, phenotype, and metabolism, which correlate with in vivo potency and persistence. In particular, though metabolic fitness is a critical quality attribute, how T cell metabolic requirements vary throughout manufacturing remains unexplored. Here, we address this limitation with optical metabolic imaging (OMI), a non-invasive, label-free method to evaluate single-cell metabolism based on autofluorescent metabolic coenzymes NAD(P)H and FAD. Using OMI, we identified the dominating impacts of media composition over the selection of antibody stimulation and/or cytokines on anti-GD2 CAR T cell metabolism, activation strength and kinetics, and phenotype. We demonstrated that OMI parameters can indicate cell cycle stage and optimal gene transfer conditions for both viral transduction and electroporation-based CRISPR/Cas9. Notably, in a virus-free CRISPR-edited anti-GD2 CAR T cell model, OMI measurements allowed accurate prediction of an oxidative metabolic phenotype that yielded higher in vivo potency against neuroblastoma. Our data supports OMI’s potential as a robust, sensitive analytical tool to identify optimal manufacturing conditions and monitor cell metabolism throughout manufacturing for increased CAR T cell yield and metabolic fitness.

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“…Inhibition of cholesterol esterification and acceleration of lipid catabolism significantly enhance the effector T cell functions and antitumor activity of CD8 + TILs 49 . In addition, TME-derived cholesterol induces TIL dysfunction by activating XBP1 50 . Consistent with this, we found that the ER-stress pathway and XBP1 expression were significantly increased during CAR-T cell expansion and disease progression in patients with PD.…”

Section: Discussionmentioning

confidence: 99%

Cholesterol efflux from C1QB-expressing macrophages is associated with resistance to chimeric antigen receptor T cell therapy in primary refractory diffuse large B cell lymphoma

Yan,

Dong,

Qiao

et al. 2024

Nat Commun

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Chimeric antigen receptor T (CAR-T) cell therapy has demonstrated promising efficacy in early trials for relapsed/refractory diffuse large B cell lymphoma (DLBCL). However, its efficacy in treating primary refractory DLBCL has not been comprehensively investigated, and the underlying resistance mechanisms remain unclear. Here, we report the outcomes of a phase I, open-label, single-arm clinical trial of relmacabtagene autoleucel (relma-cel), a CD19-targeted CAR-T cell product, with safety and efficacy as primary endpoints. Among the 12 enrolled patients, 8 experienced grade 4 hematologic toxicity of treatment-emergent adverse event. No grade ≥3 cytokine release syndrome or neurotoxicity occurred. Single-cell RNA sequencing revealed an increase proportion of C1QB-expressing macrophages in patients with progressive disease before CAR-T cell therapy. Cholesterol efflux from M2 macrophages was found to inhibit CAR-T cells cytotoxicity by inducing an immunosuppressive state in CD8+ T cells, leading to their exhaustion. Possible interactions between macrophages and CD8+ T cells, mediating lipid metabolism (AFR1-FAS), immune checkpoint activation, and T cell exhaustion (LGALS9-HAVCR2, CD86-CTLA4, and NECTIN2-TIGIT) were enhanced during disease progression. These findings suggest that cholesterol efflux from macrophages may trigger CD8+ T cell exhaustion, providing a rationale for metabolic reprogramming to counteract CAR-T treatment failure. Chinadrugtrials.org.cn identifier: CTR20200376.

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Metabolic and epigenetic orchestration of (CAR) T cell fate and function

Cited by 10 publications

References 175 publications

The inhibitory receptors PD1, Tim3, and A2aR are highly expressed during mesoCAR T cell manufacturing in advanced human epithelial ovarian cancer

The inhibitory receptors PD1, Tim3, and A2aR are highly expressed during mesoCAR T cell manufacturing in advanced human epithelial ovarian cancer

Label free metabolic imaging to enhance the efficacy of chimeric antigen receptor T cell therapy

Cholesterol efflux from C1QB-expressing macrophages is associated with resistance to chimeric antigen receptor T cell therapy in primary refractory diffuse large B cell lymphoma

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