Inhibition of AKT signaling uncouples T cell differentiation from expansion for receptor-engineered adoptive immunotherapy

Klebanoff, Christopher A.; Crompton, Joseph G.; Leonardi, Anthony; Yamamoto, Tori N.; Chandran, Smita S.; Eil, Robert L.; Sukumar, Madhusudhanan; Vodnala, Suman K.; Hu, Jinhui; Ji, Yun; Clever, David; Black, Mary A.; Gurusamy, Devikala; Kruhlak, Michael J.; Jin, Ping; Stroncek, David F.; Gattinoni, Luca; Feldman, Steven A.; Restifo, Nicholas P.

doi:10.1172/jci.insight.95103

Cited by 150 publications

(135 citation statements)

References 86 publications

(111 reference statements)

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“…Confirming previously published data, 15,16,18 AKT-inhibited T cells showed increased secondary expansion capacity after removal of the inhibitor. Moreover, we found that next to IFN-γ, AktiVIII and GDC-treated T cells also produced IL-2, which was not observed in control T cells.…”

Section: Discussionsupporting

confidence: 90%

“…However, the metabolic profiling of our AKT-inhibited T cells differed from previously reported Akt-inhibited and GSK-3β-inhibited T SCM -enriched cells. 18,33 Instead of an increased oxidative phosphorylation, we observed enhanced glycolysis in AKT-inhibited T cells, especially with AktiVIII inhibition. This observation was confirmed by the higher expression of glycolysis-associated PFKM and decreased expression of OXPHOS-associated CPT1A in our transcriptome analysis.…”

Section: Discussionmentioning

confidence: 68%

“…These data are in agreement with previously published data on AKT-inhibited T cells by us and others, 15-19 and confirms the extensive down-stream analysis by Klebanoff et al ., which highlights FOXO1 as the key player in this process. 18 With the increased expression of FOXO1 targets, increased expression of naïve-associated markers, and decreased expression of effector and apoptosis molecules, our AKT-inhibited cells possess unique and superior characteristics for adoptive cell therapy in cancer patients. However, AKT-inhibition did not preserve all naïve associated genes, and did elevated specific effector-related factors, including PRDM1, which could be involved in triggering effector cell functions.…”

Section: Discussionmentioning

confidence: 98%

“…This is in agreement with findings of Klebanoff et al ., who showed that AKT-inhibition (using low dose AktiVIII) uncouples T cell differentiation from expansion. 18 Interestingly, while expansion was not hampered by inhibiting AKT, generation of early memory cells by interference with GSK-3β clearly blocked proliferation of T cells. 15,30,33 This makes AKT a very potent candidate for the generation of effective adoptive T cell products.…”

Section: Discussionmentioning

confidence: 99%

“…This uncoupling of T cell differentiation from expansion using AKT-inhibitors has been confirmed in other models, including melanoma-derived tumor-infiltrating lymphocytes and CD19 CAR T cells, as well as by modulation of up- and down-stream targets of the AKT-pathway, including mTORC2 and PI3K-δ. 16-18,20,21 …”

Section: Introductionmentioning

confidence: 99%

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Ex vivo AKT-inhibition facilitates generation of polyfunctional stem cell memory-like CD8⁺ T cells for adoptive immunotherapy

Mousset

Hobo

et al. 2018

OncoImmunology

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Adoptive T cell therapy has shown clinical potential for patients with cancer, though effective treatment is dependent on longevity and potency of the exploited tumor-reactive T cells. Previously, we showed that ex vivo inhibition of AKT using the research compound Akt-inhibitor VIII retained differentiation and improved functionality of minor histocompatibility antigen (MiHA)-specific CD8+ T cells. Here, we compared a panel of clinically applicable AKT-inhibitors with an allosteric or adenosine triphosphate-competitive mode of action. We analyzed phenotype, functionality, metabolism and transcriptome of AKT-inhibited CD8+ T cells using different T cell activation models. Most inhibitors facilitated T cell expansion while preserving an early memory phenotype, reflected by maintenance of CD62L, CCR7 and CXCR4 expression. Moreover, transcriptome profiling revealed that AKT-inhibited CD8+ T cells clustered closely to naturally occurring stem cell-memory CD8+ T cells, while control T cells resembled effector-memory T cells. Interestingly, AKT-inhibited CD8+ T cells showed enrichment of hypoxia-associated genes, which was consistent with enhanced glycolytic function. Notably, AKT-inhibition during MiHA-specific CD8+ T cell priming uncoupled preservation of early memory differentiation from ex vivo expansion. Furthermore, AKT-inhibited MiHA-specific CD8+ T cells showed increased polyfunctionality with co-secretion of IFN-γ and IL-2 upon antigen recall. Together, these data demonstrate that AKT-inhibitors with different modality of action promote the ex vivo generation of stem cell memory-like CD8+ T cells with a unique metabolic profile and retained polyfunctionality. Akt-inhibitor VIII and GDC-0068 outperformed other inhibitors, and are therefore promising candidates for ex vivo generation of superior tumor-reactive T cells for adoptive immunotherapy in cancer patients.

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Section: Discussionsupporting

confidence: 90%

Section: Discussionmentioning

confidence: 68%

Section: Discussionmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Ex vivo AKT-inhibition facilitates generation of polyfunctional stem cell memory-like CD8⁺ T cells for adoptive immunotherapy

Mousset

Hobo

et al. 2018

OncoImmunology

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Modulation of intracellular kinase signaling to improve TIL stemness and function for adoptive cell therapy

Feng

Qiu

Shi³

et al. 2022

Cancer Medicine

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Introduction Adoptive cellular therapy with tumor‐infiltrating lymphocytes (TIL) has demonstrated promising clinical benefits in several solid tumors, but the efficacy of this therapy might be compromised by the “prone‐to‐exhaustion” phenotype of TIL and poor persistence in vivo. This calls for a robust expansion process to produce a large number of cells for clinical usage while at the same time maintaining favorable anti‐tumor function and memory phenotype. Previous studies showed that the PI3K‐AKT signaling pathway plays a key role in the regulation of T cell activation, differentiation and memory formation. Method We modulated the PI3K‐AKT pathway in TIL isolated from cervical and ovarian cancer by application of AKT or PI3K inhibitors or CRISPR knockout of AKT1 and/or AKT2, and characterized their effects on TIL phenotype and effector function. Mechanistic study was further performed with RNA‐seq analysis of AKT1/2 KO TIL in comparison to control TIL. Result The inhibition of either PI3K or AKT led to an increase in the population of effector CD8 + T cells with upregulation of activation markers, elevated CD39 − CD69 − memory T cells, and significantly enhanced cytotoxicity when cocultured with tumor cell lines and patient‐derived tumor samples. Moreover, dual knockout of AKT1 and AKT2 largely phenocopies the functional impact of AKT or PI3K inhibition on TIL. This result was further validated by RNA‐seq analysis indicating that AKT1/2 ablation primarily regulates T cell differentiation and function‐related programs. Conclusion Modulation of PI3K‐AKT signaling represents a promising strategy to enhance TIL stemness and cytotoxicity and improve the clinical outcome of current TIL‐based therapy to treat solid tumors.

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Ex vivo blockade of PI3K gamma or delta signaling enhances the antitumor potency of adoptively transferred CD8⁺ T cells

et al. 2020

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Adoptive T cell transfer therapy induces objective responses in patients with advanced malignancies. Despite these results, some individuals do not respond due to the generation of terminally differentiated T cells during the expansion protocol. As the gamma and delta catalytic subunits in the PI3K pathway are abundant in leukocytes and involved in cell activation, we posited that blocking both subunits ex vivo with the inhibitor IPI‐145 would prevent their differentiation, thereby increasing antitumor activity in vivo. However, IPI‐145 treatment generated a product with reduced antitumor activity. Instead, T cells inhibited of PI3Kγ (IPI‐549) or PI3Kδ (CAL‐101 or TGR‐1202) alone were more potent in vivo. While T cells coinhibited of PI3Kγ and PI3Kδ were less differentiated, they were functionally impaired, indicated by reduced production of effector cytokines after antigenic re‐encounter and decreased persistence in vivo. Human CAR T cells expanded with either a PI3Kγ or PI3Kδ inhibitor possessed a central memory phenotype compared to vehicle cohorts. We also found that PI3Kδ‐inhibited CARs lysed human tumors in vitro more effectively than PI3Kγ‐expanded or traditionally expanded CAR T cells. Our data imply that sole blockade of PI3Kγ or PI3Kδ generates T cells with remarkable antitumor properties, a discovery that has substantial clinical implications.

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Inhibition of AKT signaling uncouples T cell differentiation from expansion for receptor-engineered adoptive immunotherapy

Cited by 150 publications

References 86 publications

Ex vivo AKT-inhibition facilitates generation of polyfunctional stem cell memory-like CD8⁺ T cells for adoptive immunotherapy

Ex vivo AKT-inhibition facilitates generation of polyfunctional stem cell memory-like CD8⁺ T cells for adoptive immunotherapy

Modulation of intracellular kinase signaling to improve TIL stemness and function for adoptive cell therapy

Ex vivo blockade of PI3K gamma or delta signaling enhances the antitumor potency of adoptively transferred CD8⁺ T cells

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Inhibition of AKT signaling uncouples T cell differentiation from expansion for receptor-engineered adoptive immunotherapy

Cited by 150 publications

References 86 publications

Ex vivo AKT-inhibition facilitates generation of polyfunctional stem cell memory-like CD8+ T cells for adoptive immunotherapy

Ex vivo AKT-inhibition facilitates generation of polyfunctional stem cell memory-like CD8+ T cells for adoptive immunotherapy

Modulation of intracellular kinase signaling to improve TIL stemness and function for adoptive cell therapy

Ex vivo blockade of PI3K gamma or delta signaling enhances the antitumor potency of adoptively transferred CD8+ T cells

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Product

Resources

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Ex vivo AKT-inhibition facilitates generation of polyfunctional stem cell memory-like CD8⁺ T cells for adoptive immunotherapy

Ex vivo AKT-inhibition facilitates generation of polyfunctional stem cell memory-like CD8⁺ T cells for adoptive immunotherapy

Ex vivo blockade of PI3K gamma or delta signaling enhances the antitumor potency of adoptively transferred CD8⁺ T cells