Disrupting N-glycan expression on tumor cells boosts chimeric antigen receptor T cell efficacy against solid malignancies

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.

CAR T cell manufacturing from naive/stem memory T lymphocytes enhances antitumor responses while curtailing cytokine release syndrome

“…This improved therapeutic potential was associated with increased CAR T cell expansion rates ( Figure 2C ), which were evident both in the CD4 + and CD8 + compartments ( Supplemental Figure 2E ), and with a trend toward higher release of IFN-γ, especially during the second antitumor response ( Figure 2D ). In line with our previous observations ( 34 , 35 ), CD8 + CAR T cells were enriched immediately after leukemia encounter in both conditions, while CD4 + CAR T cells became prominent at later time points ( Supplemental Figure 2F ). Notably, 14 days after infusion, CAR T N/SCM contained an increased T CM percentage compared with CAR T BULK ( Figure 2E ), possibly accounting for their superior and long-lasting therapeutic activity.…”

Antigen glycosylation regulates efficacy of CAR T cells targeting CD19

“…As a result of this intrinsic alteration of surface protein glycosylation, some malignant cells may be predisposed to evade CAR binding. A recent report demonstrated this phenomenon in pancreatic cancer cells, wherein increased cellular glycosylation occurring through an unclear mechanism was associated with decreased CAR efficacy 27 . Interestingly, they found that disruption of N -acetylglucosaminyltransferase V (GnTV, encoded by MGAT5 ), a Golgi-resident glycosyltransferase that is a primary target of SPPL3 17 , improved the efficacy of CAR therapy.…”

Enhancing the anti-tumor efficacy of Bispecific T cell engagers via cell surface glycocalyx editing

“…Instead, desialylation elicits stronger immunological synapse formation between the engaged T cells and target cells. Indeed, several studies published recently revealed that bulky glycans on the surface of tumor cells result in suboptimal synapse formation induced by both CAR-T cells and BiTEs (35).…”