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A Real-time Potency Assay for Chimeric Antigen Receptor T Cells Targeting Solid and Hematological Cancer Cells
Published on: November 12, 2019
Disrupting N-glycan expression on tumor cells boosts chimeric antigen receptor T cell efficacy against solid
Beatrice Greco1,2, Valeria Malacarne3,4, Federica De Girardi1
1Innovative Immunotherapies Unit, Division of Immunology, Transplantation and Infectious Diseases, IRCCS San Raffaele Scientific Institute, 20132 Milan, Italy.
Abstract:
Immunotherapy with chimeric antigen receptor (CAR)–engineered T cells showed exceptional successes in patients with refractory B cell malignancies. However, first-in-human studies in solid tumors revealed unique hurdles contributing to poor demonstration of efficacy. Understanding the determinants of tumor recognition by CAR T cells should translate into the design of strategies that can overcome resistance. Here, we show that multiple carcinomas express extracellular N-glycans, whose abundance negatively correlates with CAR T cell killing. By knocking out mannoside acetyl-glucosaminyltransferase 5 (MGAT5) in pancreatic adenocarcinoma (PAC), we showed that N-glycans protect tumors from CAR T cell killing by interfering with proper immunological synapse formation and reducing transcriptional activation, cytokine production, and cytotoxicity. To overcome this barrier, we exploited the high metabolic demand of tumors to safely inhibit N-glycans synthesis with the glucose/mannose analog 2-deoxy-d-glucose (2DG). Treatment with 2DG disrupts the N-glycan cover on tumor cells and results in enhanced CAR T cell activity in different xenograft mouse models of PAC. Moreover, 2DG treatment interferes with the PD-1–PD-L1 axis and results in a reduced exhaustion profile of tumor-infiltrating CAR T cells in vivo. The combined 2DG and CAR T cell therapy was successful against multiple carcinomas besides PAC, including those arising from the lung, ovary, and bladder, and with different clinically relevant CAR specificities, such as CD44v6 and CEA. Overall, our results indicate that tumor N-glycosylation regulates the quality and magnitude of CAR T cell responses, paving the way for the rational design of improved therapies against solid malignancies.
Insights
Tumor N-glycans hinder chimeric antigen receptor (CAR) T cell therapy in solid tumors. Inhibiting N-glycan synthesis with 2-deoxy-d-glucose (2DG) enhances CAR T cell efficacy against carcinomas.
Area of Science:
- Oncology
- Immunology
- Biochemistry
Background:
- Chimeric antigen receptor (CAR) T cell immunotherapy has shown success in B cell malignancies but faces challenges in solid tumors.
- Tumor recognition and killing by CAR T cells are crucial for therapeutic efficacy.
- Understanding resistance mechanisms is key to improving CAR T cell therapy for solid cancers.
Purpose of the Study:
- To investigate the role of extracellular N-glycans in CAR T cell resistance in carcinomas.
- To develop strategies to overcome N-glycan-mediated CAR T cell suppression.
- To evaluate the therapeutic potential of combining N-glycan inhibition with CAR T cell therapy.
Main Methods:
- Knockout of mannoside acetyl-glucosaminyltransferase 5 (MGAT5) in pancreatic adenocarcinoma (PAC) models.
- Treatment with 2-deoxy-d-glucose (2DG) to inhibit N-glycan synthesis.
- Assessment of CAR T cell function, including synapse formation, cytotoxicity, and cytokine production.
- Evaluation of CAR T cell responses in xenograft mouse models of various carcinomas (PAC, lung, ovarian, bladder).
- Analysis of PD-1/PD-L1 expression and T cell exhaustion markers.
Main Results:
- Extracellular N-glycans on carcinomas negatively correlate with CAR T cell killing efficiency.
- MGAT5 knockout and 2DG treatment reduced tumor N-glycans, enhancing CAR T cell activity.
- 2DG treatment improved CAR T cell synapse formation, cytotoxicity, and cytokine production.
- Combined 2DG and CAR T cell therapy demonstrated efficacy in multiple solid tumor models and with different CAR specificities.
- 2DG treatment reduced T cell exhaustion by interfering with the PD-1–PD-L1 axis.
Conclusions:
- Tumor N-glycosylation is a significant determinant of CAR T cell response quality and magnitude.
- Inhibiting N-glycan synthesis with 2DG can overcome CAR T cell resistance in solid tumors.
- Combined 2DG and CAR T cell therapy represents a promising strategy for treating various carcinomas.
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