Generation of clinical-grade CD19-specific CAR-modified CD8+ memory stem cells for the treatment of human B-cell malignancies

Sabatino, Marianna; Hu, Jinhui; Sommariva, Michele; Gautam, Sanjivan; Fellowes, Vicki; Hocker, James D.; Dougherty, Sean C.; Qin, Haiying; Klebanoff, Christopher A.; Fry, Terry J.; Gress, Ronald E.; Kochenderfer, James N.; Stroncek, David F.; Ji, Yun; Gattinoni, Luca

doi:10.1182/blood-2015-11-683847

Cited by 283 publications

(285 citation statements)

References 67 publications

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“…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: 71%

“…1-3,5-9,25,29,32,34 This perspective should promote further research and clinical translation of adoptive T cell therapy with interference of the PI3K/AKT/mTOR or Wnt-signalling pathway. 15-19,21,33 Here we show that AKT-inhibition can be used for the generation of a unique T SCM -like CD8 + T cell product for adoptive transfer. Though this is irrespective of the mode of action of the inhibitor, the choice of inhibitor does influence the characteristics and, thereby, possibly the clinical potency of the therapeutic product.…”

Section: Discussionmentioning

confidence: 79%

“…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%

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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: Discussionmentioning

confidence: 71%

Section: Discussionmentioning

confidence: 79%

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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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“…The success of T-cell based CAR strategies depends on distinct T-cell subsets (6). CAR-modified CD8 + memory stem cells provide superior antitumor responses in xenograft models compared to conventional CD8 + T cells (7).…”

Section: Introductionmentioning

confidence: 99%

Intrinsic Functional Potential of NK-Cell Subsets Constrains Retargeting Driven by Chimeric Antigen Receptors

Oei

Siernicka

Graczyk-Jarzynka

et al. 2018

Cancer Immunology Research

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“…Clinical evidence is pending but preliminary results in four melanoma patients showed only limited persistence of GD2-targeted CAR T cells expanded using IL-7 and -15 [31]. Selection of more primitive starting T-cell populations prior to in vitro expansion [14] and use of signaling inhibitors that prevent effector differentiation [32] are other strategies for increasing the number of less-differentiated T cells for adoptive T-cell therapy.…”

Section: Introductionmentioning

confidence: 99%

Low interleukin-2 concentration favors generation of early memory T cells over effector phenotypes during chimeric antigen receptor T-cell expansion

et al. 2017

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Background. Adoptive T-cell therapy offers new options for cancer treatment. Clinical results suggest that T-cell persistence, depending on T-cell memory, improves efficacy. The use of interleukin (IL)-2 for in vitro T-cell expansion is not straightforward because it drives effector T-cell differentiation but does not promote the formation of T-cell memory. We have developed a cost-effective expansion protocol for chimeric antigen receptor (CAR) T cells with an early memory phenotype. Methods. Lymphocytes were transduced with third-generation lentiviral vectors and expanded using CD3/CD28 microbeads. The effects of altering the IL-2 supplementation (0-300 IU/mL) and length of expansion (10-20 days) on the phenotype of the T-cell products were analyzed. Results. High IL-2 levels led to a decrease in overall generation of early memory T cells by both decreasing central memory T cells and augmenting effectors. T memory stem cells (TSCM, CD95 + CD45RO − CD45RA + CD27 + ) were present variably during T-cell expansion. However, their presence was not IL-2 dependent but was linked to expansion kinetics. CD19-CAR T cells generated in these conditions displayed in vitro antileukemic activity. In summary, production of CAR T cells without any cytokine supplementation yielded the highest proportion of early memory T cells, provided a 10-fold cell expansion and the cells were functionally potent. Discussion. The number of early memory T cells in a T-cell preparation can be increased by simply reducing the amount of IL-2 and limiting the length of T-cell expansion, providing cells with potentially higher in vivo performance. These findings are significant for robust and cost-effective T-cell manufacturing.

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Generation of clinical-grade CD19-specific CAR-modified CD8+ memory stem cells for the treatment of human B-cell malignancies

Abstract: Key Points A platform for the generation of clinical-grade CD19-CAR–modified TSCM. CD19-CAR–modified TSCM mediate superior antitumor responses compared with CD19-CAR T cells currently used in clinical trials.

Cited by 283 publications

References 67 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

Intrinsic Functional Potential of NK-Cell Subsets Constrains Retargeting Driven by Chimeric Antigen Receptors

Low interleukin-2 concentration favors generation of early memory T cells over effector phenotypes during chimeric antigen receptor T-cell expansion

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Generation of clinical-grade CD19-specific CAR-modified CD8+ memory stem cells for the treatment of human B-cell malignancies

Abstract: Key Points A platform for the generation of clinical-grade CD19-CAR–modified TSCM. CD19-CAR–modified TSCM mediate superior antitumor responses compared with CD19-CAR T cells currently used in clinical trials.

Cited by 283 publications

References 67 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

Intrinsic Functional Potential of NK-Cell Subsets Constrains Retargeting Driven by Chimeric Antigen Receptors

Low interleukin-2 concentration favors generation of early memory T cells over effector phenotypes during chimeric antigen receptor T-cell expansion

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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