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Manufacturing Chimeric Antigen Receptor CAR T Cells for Adoptive Immunotherapy
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T cell engineered with a novel nanobody-based chimeric antigen receptor against VEGFR2 as a candidate for tumor

Fatemeh Hajari Taheri1, Mahmoud Hassani1,2, Zahra Sharifzadeh1

  • 1Department of Immunology, Hybridoma Lab, Pasteur Institute of Iran, Tehran, Iran.

IUBMB Life
|February 7, 2019
PubMed

Insights

Researchers developed a novel nanobody-based CAR T cell therapy targeting vascular endothelial growth factor receptor 2 (VEGFR2) for solid tumors. This therapy shows promise for adoptive T-cell immunotherapy against cancers dependent on angiogenesis.

Area of Science:

  • Immunology
  • Oncology
  • Biotechnology

Background:

  • Solid tumors rely on angiogenesis for growth and metastasis, making antiangiogenic therapies crucial.
  • Vascular Endothelial Growth Factor Receptor 2 (VEGFR2) is over-expressed on tumor vasculature, presenting a viable therapeutic target.

Purpose of the Study:

  • To develop and evaluate a second-generation nanobody (VHH)-based Chimeric Antigen Receptor (CAR) T cell therapy targeting VEGFR2-expressing solid tumor cells.

Main Methods:

  • Constructed a CAR T cell by linking an anti-VEGFR2 VHH to a spacer and CD28/CD3ζ signaling domains.
  • Activated T cells with anti-CD3/rIL-2 and electroporated with the CAR construct plasmid.
  • Assessed CAR T cell activation, cytokine production, degranulation, and cytotoxic activity against VEGFR2-expressing cells.

Main Results:

  • CAR T cells exhibited significant expression of activation markers (CD69, CD25) and produced substantial IL-2 and IFN-γ upon co-culture with VEGFR2-expressing cells.
  • Demonstrated increased degranulation marker (CD107a) expression and potent cytotoxic activity (>30%) against target cells.
  • Anti-VEGFR2 CAR T cells specifically lysed VEGFR2-expressing 293-KDR cells, unlike mock T cells.

Conclusions:

  • The developed anti-VEGFR2 nanobody-based CAR T cell therapy is effective in targeting and eliminating VEGFR2-expressing tumor cells.
  • This CAR T cell construct shows potential as a candidate for adoptive T-cell immunotherapy in solid tumors.
Keywords:
CAR T-cellVEGFR2chimeric antigen receptor (CAR)electroporationnanobodysolid tumor

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