Doxycycline Inducible Chimeric Antigen Receptor T Cells Targeting CD147 for Hepatocellular Carcinoma Therapy

Abstract

Chimeric antigen receptor T cell (CAR-T) therapy to hematological malignancies has demonstrated tremendous clinical outcomes. However, the therapeutic efficacy of CAR-T cells in solid tumors remains limited due to the scarcity of tumor-specific antigen targets and the poor infiltration of CAR-T cells into tumor tissue. In this study, we developed a novel inducible CAR-T cell system which targets CD147, a tumor-associated antigen for hepatocellular carcinoma (HCC). To minimize potential toxicities of CAR-T cell therapy, the Tet-On 3G system was introduced to induce CD147CAR expression in the right place at the right time. Specifically, Tet-CD147CAR lentiviral vector (LV-Tet-CD147CAR) was constructed, which comprised CD147CAR controlled by the Tet-On system. Tet-CD147CART cells were successfully generated from activated T cells by infection with LV-Tet-CD147CAR. Proliferation, cytotoxicity, and cytokine secretion of Tet-CD147CART cells were significantly increased against CD147-positive cancer cells in the presence of doxycycline (Dox) compared to Tet-CD147CART cells in the absence of Dox and PBMCs. Consistently, in vivo studies indicated that the tumor growth in nude mice was significantly inhibited by (Dox+) Tet-CD147CART cells through multiple intratumoral administration. Taken together, our results indicated that the expression and activity of CD147CAR were controlled by Dox both in vitro and in vivo, which facilitated decreased toxicity and adverse effects to CAR-T cell therapy. Moreover, this study provides viable evidence in support of the potential benefits and translation of this strategy of CAR-T cells targeting CD147 for the treatment of patients with HCC.

Immunohistochemical basigin expression level in thyroid cancer tissues

“…Some studies have provided evidence that downregulation of BSG via lentivirus vector-based RNAi decreased cell proliferation, matrigel invasion, and tumor formation in breast cancer, especially in MCF-7 breast cancer cells [61,62]. Zhang et al [63] provided evidence that chimeric antigen receptor T cells induced by doxycycline targeting BSG could be used in the treatment of liver cancer. Another study also found that doxycycline inhibited the proliferation of gallbladder cancer cells by downregulating the expression levels of BSG and induced an early apoptosis response in cancer cells [64].…”

Immunohistochemical Basigin Expression Level in Thyroid Cancer Tissues

“…Some studies have provided evidence that down-regulation of BSG via lentivirus vector-based RNAi decreased cell proliferation, matrigel invasion, and tumor formation in breast cancer, especially in MCF-7 breast cancer cells [61,62]. Zhang et al [63] provided evidence that chimeric antigen receptor T cells induced by doxycycline targeting BSG can be used in the treatment of liver cancer. Another study also found that doxycycline inhibited the proliferation of gallbladder cancer cells by down-regulating the expression levels of BSG and induced an early apoptosis response in cancer cells [64].…”

Externally-Controlled Systems for Immunotherapy: From Bench to Bedside

“…Maximal tumor lysis In vitro was assessed with 1,000 ng/ml and the prompt reversion of the CAR activity was better achieved after a short exposition (24 h) with 10 ng/ml Dox. There are also pre-clinical assays using the Tet-On 3G system in solid tumors, specifically for hepatocellular carcinoma (HCC) treatment (26). Zhang and co-workers constructed the Tet-CD147-CAR lentiviral vector to generate Tet-CD147-CART cells.…”