Combination wt-p53 and MicroRNA-125b Transfection in a Genetically Engineered Lung Cancer Model Using Dual
Meghna Talekar1, Malav Trivedi1,2, Parin Shah1
1Department of Pharmaceutical Sciences, School of Pharmacy, Bouve College of Health Sciences, Northeastern University, Boston, Massachusetts, USA.
Abstract:
Mutations in KRAS and p53 signaling pathways contribute to loss of responsiveness to current therapies and a decreased survival in lung cancer. In this study, we have investigated the delivery and transfection of wild-type (wt-) p53 and microRNA-125b (miR-125b) expressing plasmid DNA, in SK-LU-1 human lung adenocarcinoma cells as well as in Kras(G12D)/p53(fl/fl) (KP) genetically engineered mouse model of lung cancer. Systemic plasmid DNA delivery with dual CD44/EGFR-targeted hyaluronic acid (HA)-based nanoparticles (NPs) resulted in a 2- to 20-fold increase in wt-p53 and miR-125b gene expression in SK-LU-1 cells. This resulted in enhanced apoptotic activity as seen with increased APAF-1 and caspase-3 gene expression. Similarly, in vivo evaluations in KP mouse model indicated successful CD44/EGFR-targeted delivery. Tumor growth inhibition and apoptotic induction were also observed with (wt-p53+miR125b) combination therapy in KP tumor model. Lastly, J774.A1 murine macrophages co-cultured with transfected SK-LU-1 cells showed a 14- to 35-fold increase in the iNOS-Arg-1 ratio, supportive of previous results demonstrating a role of miR-125b in macrophage repolarization. Overall, these results show tremendous promise of wt-p53 and miR-125b gene therapy using dual CD44/EGFR-targeting HA NP vector for effective treatment of lung cancer.
Insights
This study developed targeted nanoparticles for lung cancer gene therapy. The dual-targeted nanoparticles successfully delivered wild-type p53 and microRNA-125b, inhibiting tumor growth and promoting apoptosis.
Area of Science:
- Oncology
- Gene Therapy
- Nanomedicine
Background:
- KRAS and p53 pathway mutations drive lung cancer resistance and poor survival.
- Effective gene delivery systems are crucial for overcoming therapeutic resistance in lung cancer.
Purpose of the Study:
- To investigate the efficacy of dual CD44/EGFR-targeted hyaluronic acid nanoparticles for delivering wild-type p53 and microRNA-125b in lung cancer.
- To evaluate the therapeutic potential of this gene therapy in both in vitro and in vivo lung cancer models.
Main Methods:
- Systemic delivery of plasmid DNA encoding wild-type p53 and microRNA-125b using dual CD44/EGFR-targeted hyaluronic acid nanoparticles.
- Transfection of SK-LU-1 human lung adenocarcinoma cells and Kras(G12D)/p53(fl/fl) genetically engineered mouse models.
- Assessment of gene expression, apoptotic activity, tumor growth inhibition, and macrophage repolarization.
Main Results:
- Nanoparticle delivery significantly increased wild-type p53 and microRNA-125b expression in cancer cells.
- Enhanced apoptotic activity (increased APAF-1 and caspase-3 expression) was observed.
- In vivo studies demonstrated successful targeted delivery, tumor growth inhibition, and apoptosis induction in the mouse model.
- Therapy also showed potential for macrophage repolarization, indicated by an increased iNOS-Arg-1 ratio.
Conclusions:
- Dual CD44/EGFR-targeted hyaluronic acid nanoparticles are a promising vector for wild-type p53 and microRNA-125b gene therapy in lung cancer.
- This combination therapy effectively inhibits tumor growth and induces apoptosis, offering a potential new treatment strategy.
- The findings support the use of this nanomedicine approach for overcoming therapeutic resistance in lung cancer.
