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Blockade of co-inhibitory receptor immune checkpoint protein TIM3/CD366 augments the anti-cancer activity of CAR-T

Binglan Zhang1, Fuping Zhu2, Song Wang1

  • 1Department of Gastroenterology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.

Genes & Diseases
|April 20, 2026
PubMed

Insights

Silencing the immune checkpoint TIM-3 in chimeric antigen receptor (CAR)-T cells enhances their ability to kill solid tumor cells. This TIM-3 blockade strategy shows promise for improving CAR-T cell therapy efficacy.

Area of Science:

  • Immunology
  • Oncology
  • Cell Therapy

Background:

  • Chimeric antigen receptor (CAR)-T cell therapy shows promise for solid tumors.
  • Immune checkpoint blockade can enhance CAR-T cell function.
  • The role of T cell immunoglobulin and mucin-domain containing-3 (TIM-3) in CAR-T cell activity is not fully understood.

Purpose of the Study:

  • To investigate the effect of TIM-3 blockade on CAR-T cell function and anti-tumor activity.
  • To construct and evaluate TIM-3-silenced CAR-T cells targeting HER2.

Main Methods:

  • Dual-promoter lentivirus vectors were used to create TIM-3-silenced CAR-T cells expressing a HER2-targeting CAR and TIM-3 shRNA.
  • Phenotypic analysis of CAR-T cells.
  • In vitro assessment of lytic cytotoxicity against target cells.
  • In vivo evaluation of anti-tumor activity in a murine xenograft model.

Main Results:

  • Down-regulation of TIM-3 did not alter the phenotype of CAR-T cells.
  • TIM-3-silenced CAR-T cells demonstrated enhanced lytic cytotoxicity in vitro.
  • TIM-3-silenced CAR-T cells exhibited robust anti-tumor activity in vivo, comparable to standard CAR-T cells.

Conclusions:

  • Down-regulating the immune checkpoint TIM-3 can enhance the anti-tumor function of CAR-T cells.
  • TIM-3 blockade offers a potential strategy to improve CAR-T cell therapy for solid tumors.
Keywords:
Cell therapyChimeric antigen receptorImmunotherapyInhibitory checkpointTIM-3

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