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Anti-Cancer Potential of Transiently Transfected HER2-Specific Human Mixed CAR-T and NK Cell Populations in
Irina O Chikileva1, Alexandra V Bruter2, Nadezhda A Persiyantseva3
1Research Institute of Experimental Therapy and Diagnostics of Tumor, NN Blokhin National Medical Center of Oncology, 115478 Moscow, Russia.
Abstract:
Human epidermal growth factor receptor 2 (HER2) is overexpressed in numerous cancer cell types. Therapeutic antibodies and chimeric antigen receptors (CARs) against HER2 were developed to treat human tumors. The major limitation of anti-HER2 CAR-T lymphocyte therapy is attributable to the low HER2 expression in a wide range of normal tissues. Thus, side effects are caused by CAR lymphocyte "on-target off-tumor" reactions. We aimed to develop safer HER2-targeting CAR-based therapy. CAR constructs against HER2 tumor-associated antigen (TAA) for transient expression were delivered into target T and natural killer (NK) cells by an effective and safe non-viral transfection method via nucleofection, excluding the risk of mutations associated with viral transduction. Different in vitro end-point and real-time assays of the CAR lymphocyte antitumor cytotoxicity and in vivo human HER2-positive tumor xenograft mice model proved potent cytotoxic activity of the generated CAR-T-NK cells. Our data suggest transient expression of anti-HER2 CARs in plasmid vectors by human lymphocytes as a safer treatment for HER2-positive human cancers. We also conducted preliminary investigations to elucidate if fucosylated chondroitin sulfate may be used as a possible agent to decrease excessive cytokine production without negative impact on the CAR lymphocyte antitumor effect.
Insights
Transiently expressed anti-HER2 CARs in T and NK cells offer a safer approach for HER2-positive cancers, minimizing on-target off-tumor effects. This method avoids viral transduction risks, enhancing therapeutic safety.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- Human epidermal growth factor receptor 2 (HER2) is a tumor-associated antigen overexpressed in various cancers.
- Current anti-HER2 CAR-T therapies face limitations due to on-target, off-tumor side effects from HER2 expression in normal tissues.
Purpose of the Study:
- To develop a safer HER2-targeting chimeric antigen receptor (CAR) therapy.
- To evaluate transient CAR expression in T and natural killer (NK) cells for improved safety and efficacy.
Main Methods:
- CAR constructs targeting HER2 were delivered into T and NK cells via non-viral nucleofection for transient expression.
- In vitro cytotoxicity assays and in vivo studies using HER2-positive tumor xenograft mouse models were performed.
- Preliminary investigation into fucosylated chondroitin sulfate's role in modulating cytokine production.
Main Results:
- Generated CAR-T-NK cells demonstrated potent cytotoxic activity against HER2-positive tumors in vitro and in vivo.
- Transient expression via nucleofection successfully delivered CAR constructs, excluding risks of viral transduction-associated mutations.
- Preliminary data suggests potential for fucosylated chondroitin sulfate to mitigate excessive cytokine release.
Conclusions:
- Transient expression of anti-HER2 CARs in human lymphocytes using plasmid vectors represents a safer therapeutic strategy for HER2-positive cancers.
- This approach mitigates "on-target off-tumor" adverse events associated with traditional CAR-T therapies.
- Further research into immunomodulatory agents like fucosylated chondroitin sulfate may enhance CAR therapy safety profiles.
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