BMC Medicine (Aug 2023)

Targeted delivery of a PD-1-blocking scFv by CD133-specific CAR-T cells using nonviral Sleeping Beauty transposition shows enhanced antitumour efficacy for advanced hepatocellular carcinoma

  • Chaopin Yang,
  • Jinqi You,
  • Qiuzhong Pan,
  • Yan Tang,
  • Liming Cai,
  • Yue Huang,
  • Jiamei Gu,
  • Yizhi Wang,
  • Xinyi Yang,
  • Yufei Du,
  • Dijun Ouyang,
  • Hao Chen,
  • Haoran Zhong,
  • Yongqiang Li,
  • Jieying Yang,
  • Yulong Han,
  • Fengze Sun,
  • Yuanyuan Chen,
  • Qijing Wang,
  • Desheng Weng,
  • Zhongqiu Liu,
  • Tong Xiang,
  • Jianchuan Xia

DOI
https://doi.org/10.1186/s12916-023-03016-0
Journal volume & issue
Vol. 21, no. 1
pp. 1 – 19

Abstract

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Abstract Background CD133 is considered a marker for cancer stem cells (CSCs) in several types of tumours, including hepatocellular carcinoma (HCC). Chimeric antigen receptor-specific T (CAR-T) cells targeting CD133-positive CSCs have emerged as a tool for the clinical treatment of HCC, but immunogenicity, the high cost of clinical-grade recombinant viral vectors and potential insertional mutagenesis limit their clinical application. Methods CD133-specific CAR-T cells secreting PD-1 blocking scFv (CD133 CAR-T and PD-1 s cells) were constructed using a sleeping beauty transposon system from minicircle technology, and the antitumour efficacy of CD133 CAR-T and PD-1 s cells was analysed in vitro and in vivo. Results A univariate analysis showed that CD133 expression in male patients at the late stage (II and III) was significantly associated with worse progression-free survival (PFS) (P = 0.0057) and overall survival (OS) (P = 0.015), and a multivariate analysis showed a trend toward worse OS (P = 0.041). Male patients with advanced HCC exhibited an approximately 20-fold higher PD-L1 combined positive score (CPS) compared with those with HCC at an early stage. We successfully generated CD133 CAR-T and PD-1 s cells that could secrete PD-1 blocking scFv based on a sleeping beauty system involving minicircle vectors. CD133 CAR-T and PD-1 s cells exhibited significant antitumour activity against HCC in vitro and in xenograft mouse models. Thus, CD133 CAR-T and PD-1 s cells may be a therapeutically tractable strategy for targeting CD133-positive CSCs in male patients with advanced HCC. Conclusions Our study provides a nonviral strategy for constructing CAR-T cells that could also secrete checkpoint blockade inhibitors based on a Sleeping Beauty system from minicircle vectors and revealed a potential benefit of this strategy for male patients with advanced HCC and high CD133 expression (median immunohistochemistry score > 2.284).

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