Defining potency of CAR<sup>+</sup> T cells: Fast and furious or slow and steady

Liadi, Ivan; Singh, Harjeet; Romain, Gabrielle; Roysam, Badrinath; Cooper, Laurence J.N.; Varadarajan, Navin

doi:10.1080/2162402x.2015.1051298

Cited by 6 publications

(9 citation statements)

References 10 publications

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“…Target cell death, pooled from four replicates, was significantly higher on microrafts with a single effector cell, displaying approximately 9.1% higher death rates over 6 h (p < 0.001; Figure 4C). CAR-T cells demonstrated the ability to kill multiple targets in series, with 2.3% ± 0.3% of single CAR-T cells killing at least two target cells, supporting previous findings that CAR-T cells can kill tumor cells in a serial manner (Figure 4D,E) [13,26,42,43]. While less than 3% of CAR-T cells participated in serial killing during the 6 h assay, longitudinal studies of 12-24 h may show higher proportions of CAR-T cells capable of killing multiple target cells.…”

Section: Microrafts Containing 3-8 Target Cells and Either One Car-t ...supporting

confidence: 86%

“…While this approach reveals the transcriptome of stimulated and non-stimulated cells, functional cytotoxicity is not measured. Microwell arrays have been used to study T cell cytotoxicity and in some cases motility; however, they do not support cell sorting and thus single cells are not available for further analysis [23,[25][26][27][28][29][30]. While these immune cell analytical tools are valuable, post-assay single cell sorting would provide the ability to correlate functionality with a variety of other properties such as gene expression or for continued expansion of cell number.…”

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confidence: 99%

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Microraft arrays for serial‐killer CD19 chimeric antigen receptor T cells and single cell isolation

et al. 2022

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Chimeric antigen receptor T (CAR‐T) cell immunotherapies have seen success in treating hematological malignancies in recent years; however, the results can be highly variable. Single cell heterogeneity plays a key role in the variable efficacy of CAR‐T cell treatments yet is largely unexplored. A major challenge is to understand the killing behavior and phenotype of individual CAR‐T cells, which are able to serially kill targets. Thus, a platform capable of measuring time‐dependent CAR‐T cell mediated killing and then isolating single cells for downstream assays would be invaluable in characterizing CAR‐T cells. An automated microraft array platform was designed to track CD19 CAR‐T cell killing of CD19+ target cells and CAR‐T cell motility over time followed by CAR‐T cell collection based on killing behavior. The platform demonstrated automated CAR‐T cell counting with up to 98% specificity and 96% sensitivity, and single cells were isolated with 89% efficiency. On average, 2.3% of single CAR‐T cells were shown to participate in serial‐killing of target cells, killing a maximum of three target cells in a 6 h period. The cytotoxicity and motility of >7000 individual CAR‐T cells was tracked across four microraft arrays. The automated microraft array platform measured temporal cell‐mediated cytotoxicity, CAR‐T cell motility, CAR‐T cell death, and CAR‐T cell to target cell distances, followed by the capability to sort any desired CAR‐T cell. The pipeline has the potential to further our understanding of T cell‐based cancer immunotherapies and improve cell‐therapy products for better patient outcomes.

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Section: Microrafts Containing 3-8 Target Cells and Either One Car-t ...supporting

confidence: 86%

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confidence: 99%

Microraft arrays for serial‐killer CD19 chimeric antigen receptor T cells and single cell isolation

et al. 2022

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“…There is increasing evidence for the comparable effectiveness of CD4+ CAR-T cells in killing target tumor cells compared to their CD8+ counterparts [ 71 – 73 ]. Although a longer conjunction period and delayed kinetics is required for CD4+ CAR-T cells, they are less prone to exhaustion or activation-induced cell death [ 74 ]. Furthermore, T cells expressing CD19-CAR, originating from a defined 1:1 ratio of CD8:CD4 CAR-T cells, have shown superior antitumor activities in vivo [ 75 , 76 ].…”

Section: Discussionmentioning

confidence: 99%

Targeting senescent cells with NKG2D-CAR T cells

Deng,

Kumar,

Xie

et al. 2024

Cell Death Discov.

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This study investigates the efficacy of NKG2D chimeric antigen receptor (CAR) engineered T cells in targeting and eliminating stress-induced senescent cells in vitro. Cellular senescence contributes to age-related tissue decline and is characterized by permanent cell cycle arrest and the senescence-associated secretory phenotype (SASP). Immunotherapy, particularly CAR-T cell therapy, emerges as a promising approach to selectively eliminate senescent cells. Our focus is on the NKG2D receptor, which binds to ligands (NKG2DLs) upregulated in senescent cells, offering a target for CAR-T cells. Using mouse embryonic fibroblasts (MEFs) and astrocytes (AST) as senescence models, we demonstrate the elevated expression of NKG2DLs in response to genotoxic and oxidative stress. NKG2D-CAR T cells displayed potent cytotoxicity against these senescent cells, with minimal effects on non-senescent cells, suggesting their potential as targeted senolytics. In conclusion, our research presents the first evidence of NKG2D-CAR T cells’ ability to target senescent brain cells, offering a novel approach to manage senescence-associated diseases. The findings pave the way for future investigations into the therapeutic applicability of NKG2D-targeting CAR-T cells in naturally aged organisms and models of aging-associated brain diseases in vivo.

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“…Several variables, including the variety of tumor-associated antigens, design of immunoreceptors, and the specific T-cell subset(s) to be modified, all collectively determine the therapeutic potency of these genetically engineered T cells. 57 A publication by Wolf and coworkers 58 has summarized various potency assays that either directly measure immune cell cytotoxic activity or use surrogate markers like cytokine release. Recently, a 3D microfluidic platform that replicates the hepatitis B virus-associated hepatocellular carcinoma (HBV-HCC) environment has been developed.…”

Section: Mps-based Assays For Testing Biological Productsmentioning

confidence: 99%

Progress in developing microphysiological systems for biological product assessment

Mansouri,

Lam,

Sung

2024

Lab Chip

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Defining potency of CAR⁺ T cells: Fast and furious or slow and steady

Cited by 6 publications

References 10 publications

Microraft arrays for serial‐killer CD19 chimeric antigen receptor T cells and single cell isolation

Microraft arrays for serial‐killer CD19 chimeric antigen receptor T cells and single cell isolation

Targeting senescent cells with NKG2D-CAR T cells

Progress in developing microphysiological systems for biological product assessment

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Defining potency of CAR+ T cells: Fast and furious or slow and steady

Cited by 6 publications

References 10 publications

Microraft arrays for serial‐killer CD19 chimeric antigen receptor T cells and single cell isolation

Microraft arrays for serial‐killer CD19 chimeric antigen receptor T cells and single cell isolation

Targeting senescent cells with NKG2D-CAR T cells

Progress in developing microphysiological systems for biological product assessment

Contact Info

Product

Resources

About

Defining potency of CAR⁺ T cells: Fast and furious or slow and steady