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Bispecific Ligand-Based EphB4 CAR-T Cells Generated Using Ephrin-B2 Show Potent Antitumor Activity Against Lung

Hirotaka Kumeda1, Koichi Hirabayashi2, Shuji Mishima1

  • 1Division of General Thoracic Surgery, Department of Surgery, Shinshu University School of Medicine, Matsumoto, Japan.

Cancer Science
|April 21, 2026
PubMed

Insights

Chimeric antigen receptor (CAR)-T cells targeting EphB4 and EphA2 show significant antitumor effects against lung adenocarcinoma. These bispecific CAR-T cells effectively reduced tumor burden and improved survival in preclinical models.

Area of Science:

  • Immunology
  • Oncology
  • Biotechnology

Background:

  • Chimeric antigen receptor (CAR)-T cell therapy shows promise for cancer treatment.
  • Ephrin B2, a ligand for Ephrin type-B receptor 4 (EphB4), has demonstrated antitumor activity.
  • The efficacy of EphB4-targeting CAR-T cells against lung adenocarcinoma requires validation.

Purpose of the Study:

  • To evaluate the dual-targeting antitumor ability of EphB4 CAR-T cells against lung adenocarcinoma.
  • To confirm the efficacy of EphB4 CAR-T cells in preclinical models.

Main Methods:

  • Immunohistochemistry was used to assess EphB4 and EphA2 expression in lung adenocarcinoma samples.
  • EphB4 CAR-T cells were generated using piggyBac-mediated gene transfer.
  • Antigen stimulation assays and co-culture experiments assessed CAR-T cell bispecificity and antitumor effects.
  • In vivo studies in mice evaluated tumor burden and survival rates.

Main Results:

  • EphB4 and EphA2 expression was high (93% and 100%) in lung adenocarcinoma samples.
  • EphB4 CAR-T cells were successfully generated with high CAR positivity and a favorable T cell phenotype.
  • EphB4 CAR-T cells demonstrated antigen-specific responses to both EphB4 and EphA2.
  • EphB4 CAR-T cells significantly suppressed lung adenocarcinoma cell growth in vitro and reduced tumor burden in vivo.
  • Mice treated with EphB4 CAR-T cells showed significantly improved survival.

Conclusions:

  • Ligand-based EphB4 CAR-T cells exhibit bispecific targeting of both EphB4 and EphA2.
  • These CAR-T cells demonstrate significant antitumor effects against lung adenocarcinoma.
  • EphB4 CAR-T cells represent a promising therapeutic strategy for lung adenocarcinoma.
Keywords:
piggyBacCAR‐T cellEphA2EphB4ephrin‐B2lung adenocarcinoma

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