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 3

αCD39 mAb drives TAEC and TAM depletion in vivo.

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αCD39 mAb drives TAEC and TAM depletion in vivo. MC38 tumor cells were i...
MC38 tumor cells were injected subcutaneously into WT C57BL/6 mice (day 0). Tumor tissues were collected on day 12 after being treated with 5 mg/kg Ab on days 8 and 11. (A) IHC images of representative MC38 tumors immunostained with anti-CD31. Scale bars: 500 μm (top) and 100 μm (bottom). (B) Quantification of microvessel density in MC38 tumor based on CD31 IHC staining. (C) Representative images of liver and heart from MC38 tumor–bearing mice stained with anti-CD31. Scale bars: 100 μm. (D) Quantification of microvessel density in liver and heart from MC38 tumor–bearing mice based on CD31 IHC staining. (E) Representative images of MC38 tumors immunostained with TUNEL and anti–Gr-1. Scale bars: 500 μm. (F) Quantitative estimates of the necrotic area in the tumor based on TUNEL staining. (G) Flow cytometry analysis of the absolute number of live cells in the tumor. (H) Representative FACS plots showing a reduction in CD45+ live cell population with high FSC and SSC after αCD39 treatment (right), consisting of mainly myeloid cells (CD3–CD11b+) (bottom right). (I) Percentage of FSChiSSChi cells in the tumor-infiltrating immune cells (TICs). (J) Quantification of the cell ratio (left) and absolute cell number (right) of TAMs (CD11b+Gr-1–F4/80hi) within tumors. (K) Quantification of MDSC (CD11b+Gr-1+) cell ratio in spleen (left) and tumor (right). Data are shown as mean ± SEM. The Mann-Whitney test was used for the statistical analysis (B, D, F, G, and I–K). Data in A–K are representative of at least 2 independent experiments. *P < 0.05; **P < 0.01. NS, not significant.