In Vivo Delivery of siRNAs Targeting EGFR and BRD4 Expression by Peptide-Modified Redox Responsive PEG-PEI Nanoparticles for the Treatment of Triple-Negative Breast Cancer.

Abstract

The study aims to investigate the in vivo distribution, antitumor effect, and safety of cell membrane-penetrating peptide-modified disulfide bond copolymer nanoparticles loaded with small-interfering RNA (siRNA) targeting epidermal growth factor receptor (EGFR) and bromodomain-containing protein 4 (BRD4) in triple-negative breast cancer (TNBC). Polyethylene glycol disulfide bond-linked polyethylenimine (PEG-SS-PEI) was modified with peptides GALA and CREKA and used as vectors to prepare siRNA nanoparticles. The GALA- and CREKA-modified PEG-SS-PEI nanoparticles (GC-NPs) were prepared by mixing siEGFR and siBRD4 (1:1) with GALA-PEG-SS-PEI and CREKA-PEG-SS-PEI (1:1) in an aqueous solution at an N/P ratio of 30:1. Nanoparticles loaded with scrambled siRNA were prepared with the same method. The gene silencing effect on EGFR and BRD4 in vitro was evaluated by Western blotting analysis. TNBC xenograft models were established by subcutaneous injection of MDA-MB-231 cells into female nude mice. At 1, 3, 6, 12, and 24 h after administration of five formulations of Cy5-siRNA (133 μg/10 g) via the tail vein, the mice were observed and imaged for a biodistribution study using an in vivo imaging system. In the pharmacodynamics experiment, tumor-bearing mice were treated with respective siRNA preparations at a dose of 133 μg/10 g for 18 days, and the body weight and tumor size were recorded every other day. The protein expression levels of EGFR, p-EGFR, PI3K, p-PI3K, Akt, p-Akt, BRD4, and c-Myc were determined using Western blotting analysis. Hematological and serum biochemical parameters, organ indices, and HE staining results for the heart, liver, spleen, lung, and kidney were analyzed to evaluate the safety of the nanoparticles. GC-NPs loaded with siEGFR and siBRD4 significantly inhibited the expression of EGFR and BRD4 in vitro. The strongest fluorescence signals were observed in the GC-NP group, especially in tumors, indicating the excellent tumor-targeted delivery of GC-NPs we constructed. Tumor growth was significantly inhibited in the GC-NP-treated group, and the expression of EGFR, p-EGFR, PI3K, p-PI3K, Akt, p-Akt, BRD4, and c-Myc in the tumors decreased by 71%, 68%, 61%, 68%, 48%, 58%, 59%, and 74% compared to the control group, respectively. There was no significant change in hematological parameters, biochemical indices, or tissue morphology in GC-NP-treated mice. SiRNA cotargeting EGFR and BRD4 delivered by GALA- and CREKA-modified PEG-SS-PEI had favorable antitumor effects in vivo toward TNBC with tumor-targeting efficacy and good biocompatibility.

• Authors: Xu Wan, Runzhou Sun, Yun Bao, Chi Zhang, Ying Wu, Yanling Gong
• Journal: Molecular pharmaceutics (Mol Pharm) Vol. 18 Issue 11 Pg. 3990-3998 (11 01 2021) ISSN: 1543-8392 [Electronic] United States
• PMID: 34591491 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• BRD4 protein, human
• Cell Cycle Proteins
• Nanoparticle Drug Delivery System
• RNA, Small Interfering
• Transcription Factors
• poly(ethylene glycol)-co-poly(ethyleneimine)
• Polyethylene Glycols
• Polyethyleneimine
• EGFR protein, human
• ErbB Receptors

Topics

• Animals
• Cell Cycle Proteins (antagonists & inhibitors, genetics)
• ErbB Receptors (antagonists & inhibitors, genetics)
• Female
• Gene Silencing
• Genetic Therapy (methods)
• Humans
• Mice
• Nanoparticle Drug Delivery System (chemistry)
• Oxidation-Reduction
• Polyethylene Glycols (chemistry)
• Polyethyleneimine (analogs & derivatives, chemistry)
• RNA, Small Interfering (administration & dosage, pharmacokinetics)
• Tissue Distribution
• Transcription Factors (antagonists & inhibitors, genetics)
• Triple Negative Breast Neoplasms (genetics, therapy)
• Xenograft Model Antitumor Assays