Glycoengineered anti-CD39 promotes anticancer responses by depleting suppressive cells and inhibiting angiogenesis in tumor models
Haohai Zhang, … , Haitao Zhao, Simon C. Robson
Haohai Zhang, … , Haitao Zhao, Simon C. Robson
Published July 1, 2022
Citation Information: J Clin Invest. 2022;132(13):e157431. https://doi.org/10.1172/JCI157431.
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Research Article Angiogenesis Immunology

Glycoengineered anti-CD39 promotes anticancer responses by depleting suppressive cells and inhibiting angiogenesis in tumor models

Abstract

Immunosuppressive cells accumulating in the tumor microenvironment constitute a formidable barrier that interferes with current immunotherapeutic approaches. A unifying feature of these tumor-associated immune and vascular endothelial cells appears to be the elevated expression of ectonucleotidase CD39, which in tandem with ecto-5′-nucleotidase CD73, catalyzes the conversion of extracellular ATP into adenosine. We glycoengineered an afucosylated anti-CD39 IgG2c and tested this reagent in mouse melanoma and colorectal tumor models. We identified major biological effects of this approach on cancer growth, associated with depletion of immunosuppressive cells, mediated through enhanced Fcγ receptor–directed (FcγR-directed), antibody-dependent cellular cytotoxicity (ADCC). Furthermore, regulatory/exhausted T cells lost CD39 expression, as a consequence of antibody-mediated trogocytosis. Most strikingly, tumor-associated macrophages and endothelial cells with high CD39 expression were effectively depleted following antibody treatment, thereby blocking angiogenesis. Tumor site–specific cellular modulation and lack of angiogenesis synergized with chemotherapy and anti–PD-L1 immunotherapy in experimental tumor models. We conclude that depleting suppressive cells and targeting tumor vasculature, through administration of afucosylated anti-CD39 antibody and the activation of ADCC, comprises an improved, purinergic system–modulating strategy for cancer therapy.

Authors

Haohai Zhang, Lili Feng, Paola de Andrade Mello, Changchuin Mao, Richard Near, Eva Csizmadia, Leo Li-Ying Chan, Keiichi Enjyoji, Wenda Gao, Haitao Zhao, Simon C. Robson

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Figure 1

CD39 is differentially expressed on tumors cell subsets, and expression level peaks on tumor-associated endothelial cells (TAECs) and tumor-associated macrophages (TAMs).

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CD39 is differentially expressed on tumors cell subsets, and expression ...
MC38 or B16F10 tumor cells were injected subcutaneously into WT C57BL/6 mice. Tissues were harvested on day 11 (MC38) and day 14 (B16F10) after tumor cell inoculation. (A) Representative images of CD39 immunohistochemistry (IHC) staining on MC38 tumor frozen sections. Scale bars: 500 μm and 50 μm (magnified views). (B) Representative images of CD39 IHC staining on B16F10 tumor frozen sections. Scale bars: 500 μm and 50 μm (magnified views). (C–F) CD39 expression on MC38 and B16F10 total tumor cells measured by flow cytometry. (C and D) Representative dot plots of CD39 expression on CD45+ and CD45– populations (left plots). CD39 expression on CD45– cells was further analyzed based on CD31 expression (right plots). (E and F) Representative histograms of CD39 expression on CD45+ population subsets, including T cells (CD3+), myeloid cells (CD3–CD11b+), and TAMs (CD3–CD11b+Gr-1–F4/80hi). (G and H) Quantification of CD39 median fluorescence intensity (MFI) on different cell subsets in MC38 (n = 6) (G) and B16F10 tumors (n = 7) (H). Data are shown as mean ± SEM. Repeated-measures 1-way ANOVA with Geisser-Greenhouse correction was used for statistical analysis. (I and J) Representative tSNE plots of the whole tumors showing the cell types and their respective CD39 expression. Data in A–J are representative of at least 2 independent experiments. *P < 0.05; ***P < 0.001; ****P < 0.0001. NS, not significant.