Antigen glycosylation regulates efficacy of CAR T cells targeting CD19

Heard, Amanda; Landmann, Jack; Hansen, Ava R; Papadopolou, Alkmini; Hsu, Yu-Sung; Selli, M; Warrington, John M.; Lattin, John; Chang, Juan; Ha, Helen; Haug-Kroeper, Martina; Doray, Balraj; Gill, Saar; Ruella, Marco; Hayer, Katharina E.; Weitzman, Matthew D.; Green, Abby M.; Fluhrer, Regina; Singh, Nathan

doi:10.1038/s41467-022-31035-7

Cited by 31 publications

(16 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Similarly, hyper-glycosylation of CD19 leads to antigen escape and CAR T-cell failure. 37 These two studies both suggest that suppressing aberrant glycosylation can improve CAR T-cell activity, and our approach that directly targets the glycome may ultimately be of complementary value, for example, in sequential or pulsed therapies that could overcome tumor glycome editing to escape one or other, but not both, of these therapeutic strategies.…”

Section: Discussionmentioning

confidence: 89%

Novel banana lectin CAR-T cells to target pancreatic tumors and tumor-associated stroma

McKenna

Ozcan

Brenner

et al. 2023

J Immunother Cancer

View full text Add to dashboard Cite

BackgroundCell therapies for solid tumors are thwarted by the hostile tumor microenvironment (TME) and by heterogeneous expression of tumor target antigens. We address both limitations with a novel class of chimeric antigen receptors based on plant lectins, which recognize the aberrant sugar residues that are a ‘hallmark’ of both malignant and associated stromal cells. We have expressed in T cells a modified lectin from banana, H84T BanLec, attached to a chimeric antigen receptor (H84T-CAR) that recognizes high-mannose (asparagine residue with five to nine mannoses). Here, we tested the efficacy of our novel H84T CAR in models of pancreatic ductal adenocarcinoma (PDAC), intractable tumors with aberrant glycosylation and characterized by desmoplastic stroma largely contributed by pancreatic stellate cells (PSCs).MethodsWe transduced human T cells with a second-generation retroviral construct expressing the H84T BanLec chimeric receptor, measured T-cell expansion, characterized T-cell phenotype, and tested their efficacy against PDAC tumor cells lines by flow cytometry quantification. In three-dimensional (3D) spheroid models, we measured H84T CAR T-cell disruption of PSC architecture, and T-cell infiltration by live imaging. We tested the activity of H84T CAR T cells against tumor xenografts derived from three PDAC cell lines. Antitumor activity was quantified by caliper measurement and bioluminescence signal and used anti-human vimentin to measure residual PSCs.ResultsH84T BanLec CAR was successfully transduced and expressed by T cells which had robust expansion and retained central memory phenotype in both CD4 and CD8 compartments. H84T CAR T cells targeted and eliminated PDAC tumor cell lines. They also disrupted PSC architecture in 3D models in vitro and reduced total tumor and stroma cells in mixed co-cultures. H84T CAR T cells exhibited improved T-cell infiltration in multicellular spheroids and had potent antitumor effects in the xenograft models. We observed no adverse effects against normal tissues.ConclusionsT cells expressing H84T CAR target malignant cells and their stroma in PDAC tumor models. The incorporation of glycan-targeting lectins within CARs thus extends their activity to include both malignant cells and their supporting stromal cells, disrupting the TME that otherwise diminishes the activity of cellular therapies against solid tumors.

show abstract

Section: Discussionmentioning

confidence: 89%

Novel banana lectin CAR-T cells to target pancreatic tumors and tumor-associated stroma

McKenna

Ozcan

Brenner

et al. 2023

J Immunother Cancer

View full text Add to dashboard Cite

show abstract

“…It is known MGAT5 is a primary target of the Golgi-resident intramembrane protease Signal peptide peptidase-like 3 (SPPL3) 26 . Along the same line as described by Greco et al ., Heard and colleagues identified expression of SPPL3 in malignant B cells as a potent regulator of resistance to CAR therapy 27 .…”

Section: Discussionmentioning

confidence: 99%

N-glycosylation engineering in chimeric antigen receptor T cells enhances anti-tumor activity

Bousser

Festjens

Meuris

et al. 2023

Preprint

View full text Add to dashboard Cite

Recently, chimeric antigen receptor (CAR) T cell technology has revolutionized cancer immunotherapy. This strategy uses synthetic CARs to redirect T cells to specific antigens expressed on the surface of tumor cells. Despite impressive progress in the treatment of hematological malignancies with CAR T cells, scientific challenges still remain for use of CAR T cell therapy to treat solid tumors. This is mainly due to the hostile tumor microenvironment and CAR-related toxicities. As the glycans decorating the T cell surface are implicated in T cell activation, differentiation, proliferation, and in the interaction of human T cells with tumor cells, we studied the role of human T cell glycosylation in more depth by manipulating their glycome. In this context, there is in vitro evidence that beta-galactoside binding lectins (Galectins) can have a strong impact on the functionality of tumor-infiltrating T cells. The high-affinity poly-LacNAc N-linked galectin ligands are mainly synthesized onto the beta1,6-GlcNAc branch introduced by N-acetylglucosaminyltransferase V (GnTV, encoded by Mgat5). We showed that knocking out Mgat5 in CD70 targeting CAR T cells leads to lower densities of poly-LacNAc modifications on the CAR T cell surface. Most interestingly, our results indicate that MGAT5 KO CD70 CAR T cells show enhanced potency to control primary tumors and relapses.

show abstract

“…SPPL3 has been noted in other screens for sensitivity to cytotoxic T cells. Loss of SPPL3 in the pre-B acute lymphoblastic leukemia (ALL) cell line NALM6 resulted in increased glycosylation of CD19 and a direct impairment of CAR T cell effector function and a reduction in their anti-tumor efficacy [31]. Another study reported that the response of CD8 T cells to HLA class I was impaired in SPPL3-KO cells through a different pathway, which involves an increase of glycosphingolipids in antigen-presenting cells.…”

Section: Discussionmentioning

confidence: 99%

Functional genomics identifies extension of complex N-glycans as a mechanism to evade lysis by natural killer cells

Zhuang

Woods

et al. 2023

Preprint

View full text Add to dashboard Cite

Somatic mutations can lead to the transformation of healthy cells into malignant cells and allow their evasion from immune surveillance. To uncover genes that play a role in the detection and lysis of tumor cells by natural killer (NK) cells, a B lymphoblastoid cell line was subjected to a genome-wide CRISPR screen. Among the top hits that facilitated NK evasion was SPPL3, which encodes an intramembrane protease that cleaves transmembrane glycosyltransferases in the Golgi apparatus. SPPL3-deficient cells accumulated glycosyltransferases, such as acetylglucosaminyltransferase 5 (MGAT5), and displayed increased N-glycosylation. Binding of NK receptors NKG2D and CD2 to their corresponding ligands MICB and CD58, and binding of rituximab to CD20, was disrupted by SPPL3-deletion. Inhibition of N-glycan maturation restored receptor binding and sensitivity to NK cells. To elucidate the mechanism of this resistant phenotype, a secondary CRISPR screen was performed in SPPL3-deficient cells. This screen identified glycosyltransferases that catalyze the formation of highly branched N-glycans and N-acetyl-lactosamine (LacNAc) extensions as key regulators that prevent killing. A significant enrichment of poly-LacNAc-containing tetra-antennary species was confirmed by glycoproteomic analysis. These findings provide mechanistic insight into how SPPL3 deletions have been linked to cancer.

show abstract

Antigen glycosylation regulates efficacy of CAR T cells targeting CD19

Cited by 31 publications

References 34 publications

Novel banana lectin CAR-T cells to target pancreatic tumors and tumor-associated stroma

Novel banana lectin CAR-T cells to target pancreatic tumors and tumor-associated stroma

N-glycosylation engineering in chimeric antigen receptor T cells enhances anti-tumor activity

Functional genomics identifies extension of complex N-glycans as a mechanism to evade lysis by natural killer cells

Contact Info

Product

Resources

About