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Polymer-Coated Extracellular Vesicles for Selective Codelivery of Chemotherapeutics and siRNA to Cancer Cells
Yong-Yu Jhan1, Guillermo Palou Zuniga1, Kanwar Abhay Singh1
1Department of Biomedical Engineering, Texas A&M University, College Station, Texas 77843-3424, United States.
Abstract:
Combination therapies involving small-interfering RNA (siRNA)-mediated gene silencing and small-molecule drugs are of high interest for cancer treatment. Among the current gene delivery carriers, cell-derived extracellular vesicles (EVs) are particularly promising candidates due to their high biocompatibility, low immunogenicity, in vivo stability, and inherent targeting ability. Here, we developed a multifunctional EV platform capable of selective codelivery of siRNA and doxorubicin (DOX) to cancer cells. siRNA was first loaded into engineered lipid-hybridized EVs (eEVs) to serve as a core. Subsequently, DOX was incorporated into a polyelectrolyte shell surrounding eEVs, which was deposited by layer-by-layer (LbL) assembly. This approach resulted in the production of a stable EV-polymer complex (LbL-eEV) with a diameter of 140.2 ± 9.0 nm and zeta potential of +22.1 ± 0.5 mV. Experiments were performed to assess cellular uptake, cytotoxicity, and gene silencing efficacy in lung adenocarcinoma cells (A549), with noncancerous fibroblast cells (CCL-210) used as a control. The results demonstrated that the LbL-eEV complex can traffic through cells and release siRNA in the cytoplasm, while delivered DOX enters nuclei to induce programmed cell death. Moreover, the inherent selectivity of the particles for cancer cells resulted in effective gene silencing and cancer killing efficiency with reduced cytotoxicity to normal cells. Synchronous delivery of siRNA and DOX was also verified by flow cytometry analysis of single cells. In summary, these data provide a proof of concept for engineering EVs to deliver multiple therapeutics and suggest that LbL-eEVs are a promising drug delivery platform for targeting cancer.
Insights
Engineered extracellular vesicles (EVs) effectively co-deliver gene silencing RNA and doxorubicin to cancer cells, enhancing treatment efficacy and reducing normal cell toxicity.
Area of Science:
- Biomedical Engineering
- Nanotechnology
- Cancer Therapeutics
Background:
- Combination therapies combining gene silencing and chemotherapy show promise for cancer treatment.
- Extracellular vesicles (EVs) offer biocompatible and stable platforms for drug delivery.
- Developing efficient carriers for simultaneous delivery of multiple therapeutic agents is crucial.
Purpose of the Study:
- To engineer a multifunctional extracellular vesicle (EV) platform for the selective co-delivery of small-interfering RNA (siRNA) and doxorubicin (DOX) to cancer cells.
- To evaluate the efficacy of this platform in vitro using lung adenocarcinoma cells.
- To demonstrate the potential of engineered EVs as a cancer drug delivery system.
Main Methods:
- Engineered EVs (eEVs) were loaded with siRNA and subsequently coated with a doxorubicin-loaded polyelectrolyte shell using layer-by-layer (LbL) assembly, forming LbL-eEV complexes.
- Characterization of LbL-eEV complex size and zeta potential.
- In vitro assessment of cellular uptake, doxorubicin release, siRNA delivery, gene silencing, and cytotoxicity in A549 lung adenocarcinoma cells and CCL-210 fibroblast cells.
Main Results:
- Stable LbL-eEV complexes (140.2 ± 9.0 nm) were successfully produced, enabling co-delivery of siRNA and DOX.
- The LbL-eEV complex demonstrated selective uptake by cancer cells, with siRNA released in the cytoplasm and DOX accumulating in the nucleus.
- Effective gene silencing and significant cancer cell death were observed, with reduced toxicity in normal cells.
Conclusions:
- Engineered EVs can be effectively utilized for the co-delivery of multiple therapeutic agents, including siRNA and small-molecule drugs.
- The LbL-eEV platform shows promise for targeted cancer therapy, offering enhanced efficacy and improved safety profiles.
- This study provides a proof of concept for advanced EV-based drug delivery systems in oncology.
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