Pre-clinical assessment of chimeric antigen receptor t cell therapy targeting CD19+ B cell malignancy

Sheng-I Hu; Ming-Chin Ko; Yi-Han Dai; Hsin-An Lin; Lih-Chyang Chen; Kuo-Yang Huang; Te-Ling Pang; Cheng-Yi Kuo; Hsin-Chung Lin

doi:10.21037/atm.2020.02.148

Pre-clinical assessment of chimeric antigen receptor t cell therapy targeting CD19+ B cell malignancy

Ann Transl Med. 2020 May;8(9):584. doi: 10.21037/atm.2020.02.148.

Authors

Sheng-I Hu¹, Ming-Chin Ko², Yi-Han Dai², Hsin-An Lin³, Lih-Chyang Chen⁴, Kuo-Yang Huang⁵, Te-Ling Pang², Cheng-Yi Kuo^{2

6}, Hsin-Chung Lin⁷

Affiliations

¹ Division of Colorectal Surgery, Department of Surgery, Tri-Service General Hospital, National Defense Medical Center, Taipei City, Taiwan.
² Uwell Biopharma Inc., New Taipei City, Taiwan.
³ Division of Infection, Department of Medicine, Tri-Service General Hospital SongShan Branch, National Defense Medical Center, Taipei City, Taiwan.
⁴ Department of Medicine, Mackay Medical College, New Taipei City, Taiwan.
⁵ Graduate Institute of Pathology and Parasitology, National Defense Medical Center, Taipei City, Taiwan.
⁶ Department and Graduate Institute of Biology and Anatomy, National Defense Medical Center, Taipei City, Taiwan.
⁷ Division of Clinical Pathology, Department of Pathology, Tri-Service General Hospital, National Defense Medical Center, Taipei City, Taiwan.

Abstract

Background: Autologous chimeric antigen receptor (CAR) T cell therapy is a promising therapeutic strategy for treating hematologic malignancies. A spectrum of serious complications caused by CAR-T cells has caught great attention. We developed a novel CAR against CD19 namely UWC19, consisting anti-CD19 single-chain variable fragment (scFv) hinged with 4-1BB and CD3z signaling domains. In this study, preclinical assessments of UWC19 were conducted to evaluate the safety and efficacy in vitro and in vivo.

Methods: To evaluate the binding activity of UWC19 cells to CD19, we measured the saturation degree of CAR with human CD19 molecules using flow cytometry in vitro. The antitumor efficacy of UWC19 cells was determined by in vitro cytotoxicity assay against CD19 positive cells and in vivo using a xenograft mouse model. Cross tissue reactivity of UWC19 cells was examined by co-culturing with cell lines from difference human tissues. Tumorigenicity was determined by subcutaneously injecting UWC19 in immunodeficient mice. Persistence was analyzed using quantitative PCR.

Results: We showed that UWC19 CAR T cells exerted highly specific binding affinity and cytotoxicity against CD19+ cells in vitro. In vivo, UWC19 CAR T cells are able to fully control disease progression in a Raji-xenografted immunodeficient mouse model. UWC19 exerted no obvious effects on the mean body mass and graft versus host disease were observed in surviving mice. We showed that UWC19 cells specifically recognized and eliminated CD19 positive cells, whereas CD19 negative cells were much less affected. No tumorigenicity of UWC19 in immunodeficient mice was observed.

Conclusions: UWC19 treatment effectively eliminated CD19 positive tumor cells with favorable toxicity profile. The findings suggest encouraging clinical prospects for its use in patients with CD19 positive B cell malignancies. Our study presented an alternative evaluation strategy for CAR-T cell products.

Keywords: CD19; Chimeric antigen receptor (CAR); UWC19; immunodeficient mice; safety evaluation; tumorigenicity.