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In Vitro Tumor Cell Rechallenge For Predictive Evaluation of Chimeric Antigen Receptor T Cell Antitumor Function
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[The Construction of ROR1 Targeting Chimeric Antigen Receptor Modified T Cells and Its Killing Effect for

Yue Chen1, Ze-Ming Mo1, Di-Yuan Qin1

  • 1Department of Cancer Center, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China.

Sichuan Da Xue Xue Bao. Yi Xue Ban = Journal of Sichuan University. Medical Science Edition
|May 21, 2019
PubMed
Abstract

Insights

Type I Receptor Tyrosine Kinase-Like Orphan Receptor (ROR1) chimeric antigen receptor T cells (CAR-T) successfully killed ROR1-expressing tumor cells in vitro. This research provides a basis for ROR1 CAR-T clinical applications.

Area of Science:

  • Immunotherapy
  • Oncology
  • Molecular Biology

Background:

  • Receptor tyrosine kinase-like orphan receptor 1 (ROR1) is a target in various cancers.
  • Chimeric antigen receptor T cell (CAR-T) therapy shows promise for cancer treatment.

Purpose of the Study:

  • To evaluate the efficacy of ROR1-specific CAR-T cells against ROR1-expressing tumors.
  • To establish the preclinical foundation for ROR1 CAR-T therapy.

Main Methods:

  • Designed and synthesized ROR1 CAR gene using lentiviral vector.
  • Confirmed CAR gene construction via double enzyme digestion.
  • Assessed ROR1 expression on tumor cell lines using flow cytometry (FCM).
  • Quantified CAR-T cell killing efficacy via FCM, LDH assay, and ELISA.
  • Measured interferon-gamma (IFN-γ) release as an indicator of T cell activation.

Main Results:

  • Successfully constructed the ROR1 CAR lentiviral vector plasmid.
  • CAR-T infection efficiency reached approximately 47.23%.
  • Multiple tumor cell lines exhibited varying degrees of ROR1 expression.
  • CAR-T cells demonstrated specific killing of ROR1-positive tumor cells.
  • Significantly higher IFN-γ release was observed from CAR-T cells compared to control T cells upon encountering ROR1-positive targets (P<0.05).

Conclusions:

  • ROR1 CAR-T cells were successfully constructed and validated.
  • CAR-T therapy specifically targets and eliminates ROR1-expressing tumor cells.
  • The observed release of IFN-γ supports the therapeutic potential of ROR1 CAR-T cells.
Keywords:
CAR-TImmunotherapyROR1

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