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Highly Efficient and Selective CAR-Gene Transfer Using CD4- and CD8-Targeted Lentiviral Vectors
Arezoo Jamali1,2, Laura Kapitza1,3, Thomas Schaser4
1Molecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, 63225 Langen, Germany.
Molecular Therapy. Methods & Clinical Development
|April 19, 2019
Summary
Vectofusin-1 enhances gene delivery for chimeric antigen receptor (CAR) T-cell therapy manufacturing. This improves receptor-targeted lentiviral vector efficiency without compromising T-cell function or selectivity.
Area of Science:
- Immunology
- Molecular Biology
- Biotechnology
Background:
- Chimeric antigen receptor (CAR) T-cell therapy shows promise for cancer treatment but faces manufacturing challenges.
- Receptor-targeted lentiviral vectors (LVs) offer selective gene delivery to T-cell subtypes, potentially improving CAR T-cell generation.
- Current receptor-targeted LVs exhibit lower gene delivery rates compared to conventional vesicular stomatitis virus (VSV)-LVs.
Purpose of the Study:
- To investigate the effect of the transduction enhancer Vectofusin-1 on gene delivery efficiency using CD4- and CD8-targeted LVs.
- To assess if Vectofusin-1 impacts the target cell selectivity and killing capability of generated CAR T-cells.
- To compare gene delivery rates mediated by receptor-targeted LVs with and without Vectofusin-1 against conventional VSV-LVs.
Main Methods:
- Utilized CD4- and CD8-targeted lentiviral vectors (LVs) encoding a CD19-CAR and a truncated low-affinity nerve growth factor receptor (ΔLNGFR) reporter.
- Administered Vectofusin-1 to enhance transduction of human T-cells.
- Evaluated gene delivery rates, target cell selectivity, and cytotoxic activity of CAR T-cells post-transduction.
- Observed early transduction signals and analyzed the cause of off-target signals.
Main Results:
- Vectofusin-1 significantly enhanced gene delivery for both CD4- and CD8-targeted LVs.
- Enhanced gene delivery was achieved without compromising T-cell selectivity or killing capability.
- Vectofusin-1 substantially reduced gene delivery rates mediated by VSV-LVs.
- An early transient off-target signal was observed, attributed to cell-bound LVs with surface-incorporated ΔLNGFR, not transgene expression in off-target cells.
Conclusions:
- Vectofusin-1 is an effective transduction enhancer for receptor-targeted LVs in CAR T-cell manufacturing.
- Receptor-targeted LVs, augmented by Vectofusin-1, can achieve gene transfer rates comparable to conventional VSV-LVs.
- This approach improves CAR T-cell generation efficiency while maintaining therapeutic efficacy and specificity.
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