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Cells of the Innate Immune Response01:28

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The innate immune response is an immediate and non-specific response against pathogens, acting swiftly to prevent the spread of infections. The primary cells involved in this response are phagocytes and natural killer (NK) cells.
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In humans, more than 80% of the genome gets transcribed. However, only around 2% of the genome codes for proteins. The remaining part produces non-coding RNAs which includes ribosomal RNAs, transfer RNAs, telomerase RNAs, and regulatory RNAs, among other types. A large number of regulatory non-coding RNAs have been classified into two groups depending upon their length – small non-coding RNAs, such as microRNA, which are less than 200 nucleotides in length, and long non-coding RNA...
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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.

International Journal of Molecular Sciences
|September 9, 2023
PubMed
Summary

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.

Keywords:
chimeric antigen receptorimmunotherapyleukemialipid nanoparticlesmRNAmultiple myelomanatural killer cellstumor

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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.