CAR-T or TCR-T cell functionality | Killing, Antigen stress assay, proliferation, Functional avidity cytokine release | Optimized, easy-to-use methods to address functionality Suitable for TCR and/or CAR candidate selection and validation of T-cell engineering approaches
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Planar glass-supported lipid bilayers, molecular imaging | | | |
Human organoids | | | |
Tumor slices | | Limited availability and viability of fresh tissue slices Readouts at single time points Allogeneic responses if using non-autologous T-cells
| Incorporation of microfluidic to improve ex vivo culture & model in vivo T infiltration into solid tumors. Optimization of culture conditions and readouts to analyze engineered T-cell functions in real-time
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Microphysiological 3D tumors | Simulates invasive growth of tumor cells Engineered -T cells enter arterial medium flow actively Assessment of T cell adherence and infiltration
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RNA sequencing | | | |
Nanostring | | | |
Polyfunctionality (Isoplexis) | | | |
Spatial RNA | | | |
Efficacy and in vivo persistence of human CAR-T and TCR-T | NSG animals | Easy engraftment of tumors and T-cells of human origin Study of human engineered T cells, prepared as those used in clinical trials, before regulatory approval
| Difficult to test combination therapies since NSG are sensitive to irradiation and chemotherapy No human TME, could skew tumor characteristics The lack of human cytokines released by innate cells limits T-cell persistence
| Use of tumor cells expressing different levels of antigen expression to mimic tumor heterogeneity Inclusion of immunosuppressive immune cells into tumors Stable expression of genes encoding for human cytokines
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| Humanized SGM3 mice | | | |
| HLA-A2 transgenic mice | | | |
| Intravital imaging | Real-time assessment of T cell trafficking and killing dynamics in vivo Characterization of functional heterogeneity within engineered-T cells in vivo Help to guide rationale choice of the composition of the infusion product
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Lymphodepletion regimens | Syngeneic models | Intact host immune system, recapitulation of the TME Prediction of on-target off-tumor toxicities Effects of lymphodepletion in epitope spreading
| Differences between mouse and human engineered T cells Engineering of murine T cells can be challenging Limited translation to clinical setting Novel combinations may not be able to be tested in mice due to lack of target-expression or toxicity
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Role of the tumor microenvironment Epitope spreading | Syngeneic models | Responsive host immunity, including TME and reactivity of endogenous, tumor-specific cells Synergy with other immunotherapies can be assessed
| Use of mouse CAR constructs. Differences between mouse and human T cells Engineering of murine T cells can be challenging
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Allogeneic host vs graft rejection | Allogeneic mixed lymphocyte reactions Proliferation Cytokine release | | | |