CAR-Ts redirected against the Thomsen-Friedenreich antigen CD176 mediate specific elimination of malignant cells from leukemia and solid tumors

Frontiers in Immunology

Immunology

Felicitas Thol¹  Gernot Beutel¹  Michael Hudecek²  Michael Hust³  Britta Maecker-Kolhoff⁴  Lisa Weiß⁵  Sarah Mayer-Hain⁵  Johanna Gellert⁵  Patrik Kehler⁵  Axel Schambach⁶  Katharina Zimmermann⁶  Sebastian Riese⁷  Rainer Blasczyk⁷  Luca Marie Beermann⁷  Britta Eiz-Vesper⁷  Agnes Bonifacius⁷  Melina Umland⁷  Anna Christina Dragon⁷  Louisa Ruhl⁷  Chiara Malinconico⁷  Fumiichiro Yamamoto⁸ 

[1] Department of Hematology, Hemostasis, Oncology and Stem Cell Transplantation, Hannover Medical School (MHH), Hannover, Germany;Department of Internal Medicine II, University Hospital of Würzburg, Wuerzburg, Germany;Department of Medical Biotechnology, Technical University of Braunschweig, Braunschweig, Germany;Department of Pediatric Hematology and Oncology, Hannover Medical School (MHH), Hannover, Germany;Glycotope GmbH, Berlin, Germany;Institute of Experimental Hematology, Hannover Medical School (MHH), Hannover, Germany;Institute of Transfusion Medicine and Transplant Engineering, Hannover Medical School (MHH), Hannover, Germany;Josep Carreras Leukaemia Research Institute (IJC), Badalona, Spain;

关键词: CD176;    Thomsen-Friedenreich antigen;    pan-tumor antigen;    carbohydrate antigen;    CAR-T cell therapy;    immunotherapy;    cancer;    solid tumors;   

DOI  :  10.3389/fimmu.2023.1219165

 received in 2023-05-08, accepted in 2023-09-28,  发布年份 2023

来源: Frontiers

【 摘 要 】

IntroductionChimeric antigen receptor-engineered T cells (CAR-Ts) are investigated in various clinical trials for the treatment of cancer entities beyond hematologic malignancies. A major hurdle is the identification of a target antigen with high expression on the tumor but no expression on healthy cells, since “on-target/off-tumor” cytotoxicity is usually intolerable. Approximately 90% of carcinomas and leukemias are positive for the Thomsen-Friedenreich carbohydrate antigen CD176, which is associated with tumor progression, metastasis and therapy resistance. In contrast, CD176 is not accessible for ligand binding on healthy cells due to prolongation by carbohydrate chains or sialylation. Thus, no “on-target/off-tumor” cytotoxicity and low probability of antigen escape is expected for corresponding CD176-CAR-Ts.MethodsUsing the anti-CD176 monoclonal antibody (mAb) Nemod-TF2, the presence of CD176 was evaluated on multiple healthy or cancerous tissues and cells. To target CD176, we generated two different 2nd generation CD176-CAR constructs differing in spacer length. Their specificity for CD176 was tested in reporter cells as well as primary CD8+ T cells upon co-cultivation with CD176+ tumor cell lines as models for CD176+ blood and solid cancer entities, as well as after unmasking CD176 on healthy cells by vibrio cholerae neuraminidase (VCN) treatment. Following that, both CD176-CARs were thoroughly examined for their ability to initiate target-specific T-cell signaling and activation, cytokine release, as well as cytotoxicity.ResultsSpecific expression of CD176 was detected on primary tumor tissues as well as on cell lines from corresponding blood and solid cancer entities. CD176-CARs mediated T-cell signaling (NF-κB activation) and T-cell activation (CD69, CD137 expression) upon recognition of CD176+ cancer cell lines and unmasked CD176, whereby a short spacer enabled superior target recognition. Importantly, they also released effector molecules (e.g. interferon-γ, granzyme B and perforin), mediated cytotoxicity against CD176+ cancer cells, and maintained functionality upon repetitive antigen stimulation. Here, CD176L-CAR-Ts exhibited slightly higher proliferation and mediator-release capacities. Since both CD176-CAR-Ts did not react towards CD176- control cells, their response proved to be target-specific.DiscussionGenetically engineered CD176-CAR-Ts specifically recognize CD176 which is widely expressed on cancer cells. Since CD176 is masked on most healthy cells, this antigen and the corresponding CAR-Ts represent a promising approach for the treatment of various blood and solid cancers while avoiding “on-target/off-tumor” cytotoxicity.

【 授权许可】

Unknown   
Copyright © 2023 Dragon, Beermann, Umland, Bonifacius, Malinconico, Ruhl, Kehler, Gellert, Weiß, Mayer-Hain, Zimmermann, Riese, Thol, Beutel, Maecker-Kolhoff, Yamamoto, Blasczyk, Schambach, Hust, Hudecek and Eiz-Vesper