Tumor-derived extracellular vesicles induce invalid cytokine release and exhaustion of CD19 CAR-T Cells

Zhu, Xiaoying; Hu, Hui; Xiao, Yi; Li, Qing; Zhong, Zhaodong; Yang, Jingmin; Ping, Zou; Cao, Yang; Meng, Fankai; Li, Wei; You, Yong; Guo, An‐Yuan; Zhu, Xiaojian

doi:10.1016/j.canlet.2022.215668

Cited by 19 publications

(16 citation statements)

References 46 publications

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“…42 It is worth noting that TEVs may induce exhaustion of CAR-T cells, impair their proliferation and anti-tumor activity. 19,[43][44][45] Specifically, the immunological checkpoints (IC) including PD-L1 found in TEVs could be responsible for the negative effect of TEVs in CAR-T cells. 19,45 Therefore, in our study, we adopted EVs derived from HEK293T cells to deliver target antigen.…”

Section: Discussionmentioning

confidence: 99%

“…The effector-memory T cell subset was reported to secrete a higher level of IFN-γ and Granzyme B. 48 , 49 Notably, chronic CD19 stimulation is considered as a major cause of T-cell dysfunction, 44 which raises concerns that the long-term exposure to CD19-EVs may lead to an exhaustion of the CART-cells. However, in the present study, the doses of CD19 EV administration were limited, and no exhaustion or dysfunction was observed after the transient stimulation of CD19 EVs both in vitro and in vivo.…”

Section: Discussionmentioning

confidence: 99%

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Extracellular Vesicles Expressing CD19 Antigen Improve Expansion and Efficacy of CD19-Targeted CAR-T Cells

Zhang¹,

Ge²,

Huang³

et al. 2023

IJN

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Background: CAR-T cell therapy is effective in the treatment of certain hematological malignancies, and the expansion and functional persistence of CAR-T cells in vivo are crucial to clinical efficacy. The aim of this study was to investigate the potential of extracellular vesicles (EVs) modified with the CAR antigen to promote the efficacy of CAR-T cells in vivo. Methods: We generated HEK293T-derived EVs to present the CD19 antigen as the CAR target. In vitro, EVs expressing CD19 antigen (CD19 EVs) were co-incubated with anti-CD19 CAR-T cells. Then, proliferation, cytokine secretion, CD107a expression, tumor killing, subsets, and immune checkpoint expression were measured to assess CAR-T cell function. After infusion of CD19 EVs pretreated CAR-T cells into a lymphoma xenograft mouse model, flow cytometry and digital PCR were used to measure the expansion of CAR-T cells, and tumor volumes were continuously monitored to assess the anti-tumor efficacy of CAR-T cells in vivo. Another mouse model was created to investigate the effect of in vivo injection of CD19 EVs on the functional persistence of CAR-T cells, and safety was determined by histopathology of the main organs. Results: CD19 EVs activated CAR-T cells in an antigen-specific and dose-dependent manner and promoted the selective expansion and cytokine secretion of co-cultured CAR-T cells. Specifically, CD19 EVs preferably increased the expansion of the CAR-T subpopulation with a high surface CD19-CAR density and consequently enhanced the anti-tumor activity of CAR-T cells. Futhermore, CD19-EVs-primed CAR-T cells achieved superior proliferation and anti-tumor effects in a mouse model with lymphoma xenograft. In vivo administration of CD19 EVs promoted the functional persistence of CAR-T cells in the xenograft mouse model. Conclusion:Our findings indicate that antigen-expressing EVs can be utilized as a boost to improve CAR-T cell efficacy in vitro and in vivo.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Extracellular Vesicles Expressing CD19 Antigen Improve Expansion and Efficacy of CD19-Targeted CAR-T Cells

Zhang¹,

Ge²,

Huang³

et al. 2023

IJN

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“…Previously, we and others demonstrated that CD19-positive vesicles can stimulate and ameliorate CAR T cells 16 or have a negative effect on functionality of T or CAR T cells [17][18][19] . Here, we showed that recognition and binding of EVs with CD19-CAR T cells relies on EVs surface antigens.…”

mentioning

confidence: 84%

CAR-tropic extracellular vesicles carry tumor-associated antigens and modulate CAR T cell functionality

Ukrainskaya

Musatova

Volkov

et al. 2023

Sci Rep

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Tumor-derived extracellular vesicles (EVs) are active contributors in metastasis and immunosuppression in tumor microenvironment. At least some of the EVs carry tumor surface molecules such as tumor-associated antigens (TAAs) and/or checkpoint inhibitors, and potentially could interact with T cells or CAR T cells. Upon contact with T cells, EVs could alter their phenotype and functions by triggering signaling through TCR or CAR reprogramming them to escape immune response. We hypothesize that EVs that possess TAA on the surface will probably interact with CAR T cells which can recognize and bind corresponding TAA. This interaction between EVs and CAR T cells may change the outcome of CAR T-based cancer immunotherapy since it should affect CAR T cells. Also, EVs could serve as adjuvants and antigenic components of antitumor vaccines. Herein, we isolated EVs from B cell precursor leukemia cell line (pre-B ALL) Nalm-6 and demonstrated that recognition and binding of CD19+EVs with CD19-CAR T cells strongly depends on the presence of CD19 antigen. CD19+EVs induce secretion of pro-inflammatory cytokines (IL-2 and IFN-y) and upregulated transcription of activation-related genes (IFNG, IFNGR1, FASLG, IL2) in CD19-CAR T cells. Tumor necrosis factor receptor superfamily (TNFRSF4 and TNFRSF9) and T-cell exhaustion markers (CTLA4, LAG3, TIM3 and PDCD1LG2) were also upregulated in CD19-CAR T cells after incubation with CD19+EVs. Long-term cultivation of CD19+ or PD-L1+EVs with CD19-CAR T cells led to increased terminal differentiation and functional exhaustion according to elevated expression of PD-1, TIGIT, CD57. In summary, our results suggest that chronic exposure of CD19-CAR T cells to CD19+EVs mediates activation and systemic exhaustion in antigen-specific manner, and this negative effect is accompanied by the impaired cytotoxic activity in vitro.

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“…Our recent studies also demonstrated that EVs secreted by tumor cells such as B-ALL could induce CD19 CAR T cell exhaustion. Mechanistically, we found that B-ALL cells secreted EVs specifically carrying CD19 antigen, and thus their persistent antigenic stimulation led to exhaustion of CD19 CAR T cells ( Zhu et al, 2022 ). In addition to EVs, tumor cells also secreted masses of metabolites to tame the TME, and consequently to regulate immunity.…”

Section: The Tme Promotes Car T Cell Exhaustionmentioning

confidence: 97%

Mechanisms of CAR T cell exhaustion and current counteraction strategies

Zhu

2022

Front. Cell Dev. Biol.

Self Cite

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The functional state of chimeric antigen receptor T (CAR T) cells determines their efficacy in vivo. Exhausted CAR T cells exhibit decreased proliferative capacity, impaired anti-tumor activity, and attenuated persistence. CAR T cell exhaustion has been recognized as a vital cause of nonresponse and relapse after CAR T cell therapy. However, the triggers and mechanisms leading to CAR T cell exhaustion remain blurry and complicated. Therefore, it is essential to clear the regulation network of CAR T cell exhaustion and explore potent solutions. Here, we review the diverse inducers of CAR T cell exhaustion in terms of manufacture process and immunosuppressive tumor microenvironment. In addition to the admitted immune checkpoint blockade, we also describe promising strategies that may reverse CAR T cell exhaustion including targeting the tumor microenvironment, epigenetics and transcriptomics.

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Tumor-derived extracellular vesicles induce invalid cytokine release and exhaustion of CD19 CAR-T Cells

Cited by 19 publications

References 46 publications

Extracellular Vesicles Expressing CD19 Antigen Improve Expansion and Efficacy of CD19-Targeted CAR-T Cells

Extracellular Vesicles Expressing CD19 Antigen Improve Expansion and Efficacy of CD19-Targeted CAR-T Cells

CAR-tropic extracellular vesicles carry tumor-associated antigens and modulate CAR T cell functionality

Mechanisms of CAR T cell exhaustion and current counteraction strategies

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