Bispecific CAR-T Cells Targeting FAP and GPC3 Have the Potential to Treat Hepatocellular Carcinoma

Chimeric antigen receptor (CAR) T cell therapy has demonstrated robust efficacy fi cacy against hematological malignancies, but there are still some challenges regarding treating solid tumors, including tumor heterogeneity, antigen escape, and an immunosuppressive microenvironment. Here, we found that SNU398, a hepatocellular carcinoma (HCC) cell line, exhibited high expression levels of fi broblast activation protein (FAP) and Glypican 3 (GPC3), which were negatively correlated with patient prognosis. The HepG2 HCC cell line highly expressed GPC3, while the SNU387 cell line exhibited high expression of FAP. Thus, we developed bispecific fi c CAR-T cells to simultaneously target FAP and GPC3 to address tumor heterogeneity in HCC. The anti-FAP-GPC3 bispecific fi c CAR-T cells could recognize and be activated by FAP or GPC3 expressed by tumor cells. Compared with anti-FAP CAR-T cells or antiGPC3 CAR-T cells, bispecific fi c CAR-T cells achieved more robust activity against tumor cells expressing FAP and GPC3 in vitro. The anti-FAP-GPC3 bispecific fi c CAR-T cells also exhibited superior antitumor efficacy fi cacy and significantly fi cantly prolonged the survival of mice compared with single-target CAR-T cells in vivo. Overall, the use of anti-FAP-GPC3 bispecific fi c CAR-T cells is a promising treatment approach to reduce tumor recurrence caused by tumor antigen heterogeneity.