Preclinical Activity of Embryonic Annexin A2-Specific Chimeric Antigen Receptor T Cells Against Ovarian Cancer

Leonard Leong , Heng Liang Tan , Simeon Cua , Kylie Su Mei Yong , Qingfeng Chen , Andre Choo 
Source: Int J Mol Sci
Publication Date: (2020)
Issue: 21(2): 381
Research Area:
Cancer Research/Cell Biology
Immunotherapy / Hematology
Cells used in publication:
T cell, human peripheral blood unstim.
Species: human
Tissue Origin: blood
4D-Nucleofector® X-Unit

Nucleofection of mRNA into T cells was carried out on a 4D-Nucleofector system with the P3 Primary Cell 4D-Nucleofector®® X Kit according to the manufacturer’s protocol. Briefly, 1×106 or 5×106 T cells were mixed with equimolar concentrations of mRNA before nucleofection using program EO-115. Nucleofected T cells were immediately utilised in functional assays


Chimeric antigen receptors (CARs) have found clinical success in B cell malignancies, but a dearth of potential targets limits their wider clinical application, especially in solid tumours. Here, we describe the development of an anti-annexin A2 CAR, CAR(2448), derived from an antibody found to have activity against epithelial ovarian cancer cell lines. The spacer length of CAR(2448) was optimised based on in vitro cytotoxic activity against ovarian cancer (OC) cell lines via a real-time cytotoxicity assay. The longer spacer CAR(2448)L T cells exhibit significant effector activity, inducing inflammatory cytokine release and cytotoxicity against OC cell lines. Furthermore, CAR(2448)L-BBz T cells induced enhanced survival in an in vivo OC xenograft model and reduced tumour volume by 76.6%. Our preclinical studies of CAR(2448) suggest its potential for the unmet need of novel strategies for the treatment of ovarian cancer.