p40-engineered CAR T-cells targeting CD276 enhance anti-tumor efficacy in non-small cell lung cancer

Pak J Pharm Sci. 2026 Aug;39(8):2352-2359. doi: 10.36721/PJPS.2026.39.8.221.1.

Abstract

Background: Chimeric antigen receptor (CAR) T-cells have shown remarkable therapeutic efficacy in hematological malignancies; however, effective breakthroughs in solid tumors have not yet been achieved.

Objectives: This study aimed to explore the therapeutic potential of p40-engineered CAR T-cells targeting CD276 (p40-H3BBz) for the treatment of non-small cell lung cancer (NSCLC).

Methods: A CAR targeting CD276 (H3BBz) was engineered to co-express the p40 subunit via lentiviral transduction. The function of p40-H3BBz was evaluated by measuring cytokine secretion levels and performing luciferase-based cytotoxicity assays in vitro. In vivo efficacy was assessed using tumor-bearing mouse models, and T-cell infiltration was analyzed by immunohistochemistry.

Results: The transduction efficiencies of H3BBz and p40-H3BBz were 63.7% and 52.3%, respectively, with both populations predominantly composed of CD4+ T-cells. Upon activation, p40-H3BBz exhibited increased IL-23 secretion, while IL-12 levels were comparable to those of H3BBz in vitro. p40-H3BBz also demonstrated enhanced cytotoxicity, degranulation, and increased production of IL-23 and IFN-γ. In vivo, p40-H3BBz CAR T-cells showed superior tumor growth inhibition, increased T-cell infiltration, and elevated IL-23 levels within tumors, without inducing significant toxicity in major organs.

Conclusion: p40-H3BBz exhibits enhanced cytokine release and cytotoxic activity, representing a promising and safe therapeutic strategy for the treatment of NSCLC.

Keywords: CD276 antigen; Chimeric antigen receptor; Interleukin 23; Non-small cell lung cancer.

MeSH terms

  • Animals
  • Carcinoma, Non-Small-Cell Lung* / immunology
  • Carcinoma, Non-Small-Cell Lung* / pathology
  • Carcinoma, Non-Small-Cell Lung* / therapy
  • Cell Line, Tumor
  • Female
  • Humans
  • Immunotherapy, Adoptive* / methods
  • Lung Neoplasms* / immunology
  • Lung Neoplasms* / pathology
  • Lung Neoplasms* / therapy
  • Mice
  • Receptors, Chimeric Antigen* / genetics
  • Receptors, Chimeric Antigen* / immunology
  • T-Lymphocytes* / immunology
  • Xenograft Model Antitumor Assays

Substances

  • Receptors, Chimeric Antigen