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Updated: Aug 24, 2025
Testing the In Vitro and In Vivo Efficiency of mRNA-Lipid Nanoparticles Formulated by Microfluidic Mixing
Published on: January 20, 2023
Non-viral inducible caspase 9 mRNA delivery using lipid nanoparticles against breast cancer: An in vitro study
Ikumi Nakashima1, Shoji Saito2, Eiichi Akahoshi3
1Department of Pediatrics, Shinshu University School of Medicine, Matsumoto, Japan.
Abstract:
Breast cancer is a complex heterogeneous disease with unique molecular subtypes, which limits the development of optimized treatment strategies for each subtype. Cancer gene therapy and potential therapeutics for advanced/refractory cancers can be promising for breast cancer. Combining tumor-tropic lipid nanoparticles (LNPs) and inducible caspase-9 (iC9) mRNA, we aimed to develop a novel treatment strategy for refractory breast cancer. LNP's anti-tumor effects were tested in vitro in three breast cancer cell lines: MDA-MB231, SKBR3, and MCF-7. Tumor cells were treated with LNPs encapsulated with eGFP or iC9 mRNA and chemical inducers of dimerization (CID). Apoptosis-related genes were evaluated by reverse transcriptase quantitative PCR. LNPs could efficiently deliver encapsulated GFP mRNA to all three cancer cell lines (>80% GFP expression. in target cells). Furthermore, LNPs encapsulated with iC9 mRNA (iC9-LNPs) and CID showed cytotoxic activity against all cancer cell lines in vitro. Interestingly, susceptibility to iC9 gene therapy was heterogeneous among cancer cell lines. iC9-LNPs with CID-induced potent cytotoxic effects against SKBR3 and MDA-MB231 cells, but only a mild cytotoxic effect on MCF7 cells. Quantification of apoptosis-related genes suggested that a high BAX/Bcl-2 ratio might be associated with iC9-LNP + CID susceptibility. Thus, cancer gene therapy using iC9-LNPs and CID could be a promising alternative for the treatment of breast cancers, especially for aggressive breast cancers.
Insights
Novel lipid nanoparticles (LNPs) delivering inducible caspase-9 (iC9) mRNA show promise for refractory breast cancer treatment. This gene therapy approach, combined with chemical inducers of dimerization (CID), demonstrated potent anti-cancer effects in vitro.
Area of Science:
- Oncology
- Gene Therapy
- Nanotechnology
Background:
- Breast cancer's heterogeneity complicates treatment strategies.
- Advanced or refractory breast cancers require novel therapeutic approaches.
- Cancer gene therapy offers a promising avenue for challenging cases.
Purpose of the Study:
- To develop and evaluate a novel gene therapy for refractory breast cancer.
- To investigate the efficacy of tumor-tropic lipid nanoparticles (LNPs) carrying inducible caspase-9 (iC9) mRNA.
- To assess the combined effect of iC9-loaded LNPs and chemical inducers of dimerization (CID).
Main Methods:
- Lipid nanoparticles (LNPs) were engineered to encapsulate eGFP or iC9 mRNA.
- In vitro studies were conducted on three breast cancer cell lines (MDA-MB231, SKBR3, MCF-7).
- Apoptosis-related gene expression was analyzed using reverse transcriptase quantitative PCR.
Main Results:
- LNPs efficiently delivered GFP mRNA to all tested cancer cell lines (>80% expression).
- iC9-loaded LNPs combined with CID exhibited significant cytotoxic activity against SKBR3 and MDA-MB231 cells.
- MCF-7 cells showed milder sensitivity, suggesting heterogeneous responses to iC9 gene therapy.
Conclusions:
- Cancer gene therapy using iC9-loaded LNPs and CID is a potential treatment for refractory breast cancers.
- A high BAX/Bcl-2 ratio may correlate with increased susceptibility to this gene therapy.
- This approach holds promise, particularly for aggressive breast cancer subtypes.
