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c-JUN enhances CRISPR knockin anti-B7-H3 CAR T cell function in small cell lung cancer and thoracic SMARCA4-deficient

Hyatt Balke-Want1, Vimal Keerthi2, Maria Del Carmen Arenas2

  • 1Center for Cancer Cell Therapy, Stanford Cancer Institute, Stanford University, Stanford, CA, USA; Department I of Internal Medicine, Center for Integrated Oncology Aachen Bonn Cologne Duesseldorf, University of Cologne, Medical Faculty and University Hospital Cologne, Cologne, Germany; Cancer Center Cologne Essen (CCCE), Cologne, Germany; Center for Molecular Medicine Cologne (CMMC), University of Cologne, Cologne, Germany.

Cell Reports. Medicine
|January 21, 2026
PubMed

Insights

Chimeric antigen receptor (CAR) T cells targeting B7-H3 show promise against small cell lung cancer (SCLC) and related tumors. Engineered T cells with c-JUN overcome tumor suppression, enhancing cancer cell killing and enabling scalable manufacturing.

Area of Science:

  • Oncology
  • Immunology
  • Gene Therapy

Background:

  • Small cell lung cancer (SCLC) is aggressive and limits T cell responses by downregulating major histocompatibility (MHC) class I molecules.
  • Chimeric antigen receptor (CAR) T cells offer a potential MHC-independent therapeutic strategy for SCLC, but suitable targets are scarce.
  • B7-H3/CD276 is identified as a target antigen expressed on SCLC and thoracic SMARCA4-deficient undifferentiated tumors (UTs).

Purpose of the Study:

  • To evaluate B7-H3 as a CAR T cell target for SCLC and SCLC-mimicking thoracic SMARCA4-deficient UTs.
  • To engineer CAR T cells to overcome tumor-induced immunosuppression mediated by transforming growth factor β1 (TGF-β1).
  • To assess the efficacy and manufacturing feasibility of novel CAR T cell constructs.

Main Methods:

  • Expression analysis of B7-H3 in SCLC and thoracic SMARCA4-deficient UTs.
  • Development of B7-H3-specific CAR T cells.
  • CRISPR-Cas9 gene editing to knock in c-JUN and B7-H3 CAR into the TRAC locus of human T cells.
  • Assessment of CAR T cell killing activity against SCLC and UT cell lines in vitro.
  • Evaluation of transforming growth factor β1 (TGF-β1) mediated immunosuppression.
  • Good manufacturing practice (GMP) clinical-scale manufacturing assessment.

Main Results:

  • B7-H3 is expressed on SCLC and thoracic SMARCA4-deficient UTs.
  • Thoracic SMARCA4-deficient UTs suppress B7-H3 CAR T cell killing via TGF-β1 secretion.
  • Engineered c-JUN+B7-H3 CAR T cells demonstrate enhanced killing of SCLC with low antigen density and thoracic SMARCA4-deficient UTs.
  • Non-viral, CRISPR-Cas9 mediated engineering of c-JUN+B7-H3 CAR T cells is feasible.
  • GMP clinical-scale manufacturing of c-JUN+B7-H3 CAR T cells is achievable.

Conclusions:

  • B7-H3 is a viable target for CAR T cell therapy in SCLC and related thoracic tumors.
  • Co-expression of c-JUN enhances CAR T cell efficacy against immunosuppressive tumors.
  • Engineered c-JUN+B7-H3 CAR T cells represent a promising therapeutic platform for aggressive thoracic malignancies.
  • The developed CAR T cell therapy is amenable to clinical-scale GMP manufacturing.
Keywords:
B7-H3CRISPR knock-incell therapy scalingnon-viral CAR T-cellssmall cell lung cancerthoracic SMARCA4-deficient undifferentiated tumortransforming growth factor-beta 1

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