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Stealth magnetic nanocarriers of siRNA as platform for breast cancer theranostics
J Bruniaux1, S Ben Djemaa1, K Hervé-Aubert1
1Université François-Rabelais, EA6295 « Nanomédicaments et Nanosondes », Tours, 37200, France.
Abstract:
The endogenous mechanism of RNA interference is more and more used in research to obtain specific down-regulation of gene expression in diseases such as breast cancer. Currently, despite the new fields of study open up by RNA interference, the rapid degradation of siRNA by nucleases and their negative charges prevent them from crossing cell membranes. To overcome these limitations, superparamagnetic iron oxide nanoparticles (SPIONs) represent a promising alternative for nucleic acid delivery. Previously, we reported the magnetic siRNA nanovectors (MSN) formulation using electrostatic assembly of (1) SPIONs, also able to act as contrast agents for magnetic resonance imaging (MRI), (2) siRNA and (3) chitosan aiming at their protection and enhancing their transfection efficacy. However, these nanoparticles displayed low stability in biological suspensions and inefficient transfection of active siRNA. This work aimed at upgrading MSN to Stealth MSN (S-MSN) by adding a polyethylene glycol coating to ensure colloidal stability and stealth properties. Furthermore, another polymer (poly-L-arginine) was added for efficient siRNA transfection and the quantitative composition of the formulation was adapted for biological purposes. Results showed that S-MSN provide high siRNA complexation and protection against enzymatic degradation. Green fluorescent protein (GFP) specific down-regulation on MDA-MB231/GFP cells was comparable to that of commercially available reagents, without observable cytotoxicity. According to our works, S-MSN appears as an effective formulation for in vitro siRNA specific delivery.
Insights
Stealth magnetic siRNA nanovectors (S-MSN) overcome limitations in gene silencing by enhancing stability and cellular uptake. This advanced formulation shows effective in vitro delivery for potential breast cancer therapies.
Area of Science:
- Biotechnology
- Nanomedicine
- Molecular Biology
Background:
- RNA interference (RNAi) is crucial for gene expression regulation in diseases like breast cancer.
- Short interfering RNA (siRNA) faces challenges like nuclease degradation and poor cell membrane penetration.
- Superparamagnetic iron oxide nanoparticles (SPIONs) offer potential for nucleic acid delivery and imaging.
Purpose of the Study:
- To develop improved magnetic siRNA nanovectors (S-MSN) with enhanced stability and transfection efficiency.
- To address limitations of previous magnetic siRNA nanovectors (MSN) including poor stability and transfection.
- To create a formulation suitable for in vitro siRNA delivery in cancer research.
Main Methods:
- Formulation of Stealth MSN (S-MSN) using SPIONs, siRNA, chitosan, polyethylene glycol (PEG), and poly-L-arginine.
- Evaluation of S-MSN colloidal stability and siRNA complexation.
- Assessment of siRNA protection against enzymatic degradation.
- In vitro transfection efficacy and cytotoxicity studies using MDA-MB231/GFP cells.
Main Results:
- S-MSN demonstrated high siRNA complexation and significant protection against enzymatic degradation.
- PEGylation improved colloidal stability and stealth properties of the nanovectors.
- Achieved efficient, specific gene down-regulation of Green fluorescent protein (GFP) in cancer cells.
- No observable cytotoxicity was detected with the S-MSN formulation.
Conclusions:
- S-MSN represent an effective and stable formulation for in vitro siRNA delivery.
- The developed nanovectors show promise for targeted gene silencing applications in diseases like breast cancer.
- S-MSN offer a viable alternative to existing siRNA delivery methods with improved performance.
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