Related Experiment Video
Updated: Jul 16, 2025
Advances in Human Induced Pluripotent Stem Cell-Derived Chimeric Antigen Receptor-Expressing Natural Killer Cells
Published on: February 14, 2025
CAR-NK Cells Generated with mRNA-LNPs Kill Tumor Target Cells In Vitro and In Vivo
Vita Golubovskaya1, John Sienkiewicz1, Jinying Sun1
1Promab Biotechnologies, 2600 Hilltop Drive, Richmond, CA 94806, USA.
Abstract:
Natural killer (NK) cells are cytotoxic lymphocytes that are critical for the innate immune system. Engineering NK cells with chimeric antigen receptors (CARs) allows CAR-NK cells to target tumor antigens more effectively. In this report, we present novel CAR mRNA-LNP (lipid nanoparticle) technology to effectively transfect NK cells expanded from primary PBMCs and to generate functional CAR-NK cells. CD19-CAR mRNA and BCMA-CAR mRNA were embedded into LNPs that resulted in 78% and 95% CAR expression in NK cells, respectively. BCMA-CAR-NK cells after transfection with CAR mRNA-LNPs killed multiple myeloma RPMI8226 and MM1S cells and secreted IFN-gamma and Granzyme B in a dose-dependent manner in vitro. In addition, CD19-CAR-NK cells generated with CAR mRNA-LNPs killed Daudi and Nalm-6 cells and secreted IFN-gamma and Granzyme B in a dose-dependent manner. Both BCMA-CAR-NK and CD19-CAR-NK cells showed significantly higher cytotoxicity, IFN-gamma, and Granzyme B secretion compared with normal NK cells. Moreover, CD19-CAR-NK cells significantly blocked Nalm-6 tumor growth in vivo. Thus, non-viral delivery of CAR mRNA-LNPs can be used to generate functional CAR-NK cells with high anti-tumor activity.
Insights
Novel lipid nanoparticle (LNP) technology effectively delivers chimeric antigen receptor (CAR) mRNA to natural killer (NK) cells. This generates functional CAR-NK cells with potent anti-tumor activity, offering a promising non-viral immunotherapy approach.
Area of Science:
- Immunology
- Cell Biology
- Biotechnology
Background:
- Natural killer (NK) cells are vital cytotoxic lymphocytes of the innate immune system.
- Engineering NK cells with chimeric antigen receptors (CARs) enhances their tumor-targeting capabilities.
Purpose of the Study:
- To develop and evaluate a novel CAR mRNA-lipid nanoparticle (LNP) delivery system for generating functional CAR-NK cells.
- To assess the in vitro and in vivo anti-tumor efficacy of CAR-NK cells produced via this non-viral method.
Main Methods:
- Primary peripheral blood mononuclear cells (PBMCs) were expanded and transfected with CD19-CAR or BCMA-CAR mRNA encapsulated in LNPs.
- CAR expression levels, in vitro cytotoxicity against cancer cell lines, and cytokine secretion (IFN-gamma, Granzyme B) were analyzed.
- In vivo anti-tumor efficacy of CD19-CAR-NK cells was evaluated in a tumor xenograft model.
Main Results:
- High CAR expression was achieved (78% for CD19-CAR, 95% for BCMA-CAR) using the CAR mRNA-LNP technology.
- Transfected BCMA-CAR-NK and CD19-CAR-NK cells demonstrated potent dose-dependent killing of multiple myeloma and leukemia cell lines, respectively.
- CAR-NK cells exhibited significantly enhanced cytotoxicity and cytokine secretion compared to untransfected NK cells, with CD19-CAR-NK cells showing significant in vivo tumor growth inhibition.
Conclusions:
- Non-viral delivery of CAR mRNA via LNPs is an effective method for generating functional CAR-NK cells.
- This approach yields CAR-NK cells with high anti-tumor activity, demonstrating potential for advanced cancer immunotherapies.
More Related Videos
Related Concept Videos
01:28Cells of the Innate Immune Response
Phagocytes
Phagocytes police the peripheral tissues by removing cellular debris and responding to the invasion of foreign substances or pathogens. Many phagocytes attack and remove microorganisms even before lymphocytes detect them. The human body has two general...
02:39lncRNA - Long Non-coding RNAs