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Published on: July 22, 2019
CRISPR-Cas9 engineering of CAR-T cells: Can non-viral nanoparticles unlock safer and scalable genome editing?
Bruna My1, Andrea Lia2,3,4, Ludovica Rizzo1
1Department of Mathematics and Physics "Ennio De Giorgi", University of Salento, c/o Campus Ecotekne, Lecce, Italy.
Abstract:
CAR-T cell therapy has revolutionized the treatment of hematologic malignancies. Still, durable activity in tumors remains limited by antigen heterogeneity and escape, immunosuppressive tumor microenvironment, and restricted persistence. Genome engineering with CRISPR-Cas systems offers a powerful route to reprogram CAR-T cells; however, translation increasingly depends on how editing payloads are delivered. Viral vectors remain a benchmark for efficient gene transfer, cargo constraints, insertional risk, immunogenicity, and manufacturing complexity motivate the development of safer, more scalable non-viral platforms. In this review, we provide an overview of CAR designs, clinical use, and current ex vivo manufacturing workflow; compare viral and non-viral delivery routes while distinguishing established ex vivo editing from emerging in vivo T cell programming; and outline genome-engineering strategies organized by therapeutic goals. We highlight feasibility trade-offs and discuss how nanoparticles could enable transient, non-viral delivery of genome editors, while noting that robust T cell targeting and standardized potency/safety assays remain key bottlenecks.
Insights
Genome engineering advances CAR-T cell therapy for blood cancers. Nanoparticles offer a promising non-viral method for delivering CRISPR-Cas editing tools, overcoming limitations of current viral vectors.
Area of Science:
- Immunology
- Molecular Biology
- Biotechnology
Background:
- Chimeric antigen receptor (CAR)-T cell therapy has transformed hematologic malignancy treatment.
- Challenges include antigen escape, tumor microenvironment suppression, and limited CAR-T cell persistence.
Purpose of the Study:
- To review CAR designs, clinical applications, and manufacturing.
- To compare viral and non-viral delivery systems for CAR-T cell genome engineering.
- To discuss strategies for improving CAR-T cell efficacy and persistence.
Main Methods:
- Review of current literature on CAR-T cell therapy, genome engineering, and delivery systems.
- Comparison of viral vectors versus non-viral platforms, including nanoparticle-based approaches.
- Analysis of genome engineering strategies for therapeutic goals.
Main Results:
- Viral vectors are effective but have limitations (safety, manufacturing).
- Non-viral platforms, particularly nanoparticles, show potential for transient, targeted delivery of genome editors.
- Key bottlenecks include T cell targeting and standardized potency/safety assays.
Conclusions:
- Genome engineering holds significant promise for enhancing CAR-T cell therapy.
- Non-viral delivery methods like nanoparticles are crucial for future development.
- Further research is needed to address targeting and assay standardization for clinical translation.
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