GL67 lipid-based liposomal formulation for efficient siRNA delivery into human lung cancer cells
Somayah J Jarallah1, Ahmad M Aldossary2, Essam A Tawfik1
1Advanced Diagnostics and Therapeutics Institute, Health Sector, King Abdulaziz City for Science and Technology (KACST), Riyadh 11442, Saudi Arabia.
Abstract:
The efficient delivery of small interfering RNA (siRNA) to the targeted cells significantly affects the regulation of the overexpressed proteins involved in the progression of several genetic diseases. SiRNA molecules in naked form suffer from low internalization across the cell membrane, high susceptibility to degradation by nuclease enzyme and low stability, which hinder their efficacy. Therefore, there is an urge to develop a delivery system that can protect siRNA from degradation and facilitate their uptake across the cell membrane. In this study, the cationic lipid (GL67) was exploited, in addition to DC-Chol and DOPE lipids, to design an efficient liposomal nanocarrier for siRNA delivery. The physiochemical characterizations demonstrated that the molar ratio of 3:1 has proper particle size measurements from 144 nm to 332 nm and zeta potential of -9 mV to 47 mV that depends on the ratio of the GL67 in the liposomal formulation. Gel retardation assay exhibited that increasing the percentage of GL67 in the formulations has a good impact on the encapsulation efficiency compared to DC-Chol. The optimal formulations of the 3:1 M ratio also showed high metabolic activity against A549 cells following a 24 h cell exposure. Flow cytometry findings showed that the highest GL67 lipid ratio (100 % GL67 and 0 % DC-Chol) had the highest percentage of cellular uptake. The lipoplex nanocarriers based on GL67 lipid could potentially influence treating genetic diseases owing to the high internalization efficiency and safety profile.
Insights
This study developed a novel liposomal nanocarrier using GL67 cationic lipid for enhanced small interfering RNA (siRNA) delivery. The GL67-based system demonstrated superior cellular uptake and safety, offering potential for genetic disease treatment.
Area of Science:
- Biotechnology
- Nanomedicine
- Molecular Biology
Background:
- Small interfering RNA (siRNA) is crucial for regulating overexpressed proteins in genetic diseases.
- Naked siRNA faces challenges like poor cell membrane internalization, degradation, and instability, limiting therapeutic efficacy.
- Effective delivery systems are needed to protect siRNA and enhance cellular uptake.
Purpose of the Study:
- To design an efficient liposomal nanocarrier for siRNA delivery using GL67, DC-Chol, and DOPE lipids.
- To characterize the physiochemical properties and evaluate the efficacy of the developed nanocarrier system.
Main Methods:
- Liposomal nanocarriers were formulated using GL67, DC-Chol, and DOPE lipids at a 3:1 molar ratio.
- Physiochemical characterization included particle size, zeta potential, and gel retardation assays.
- Cellular uptake and metabolic activity were assessed using flow cytometry and cell exposure assays on A549 cells.
Main Results:
- Liposomal formulations exhibited particle sizes from 144-332 nm and zeta potentials from -9 mV to 47 mV.
- Increasing GL67 content improved encapsulation efficiency and demonstrated high metabolic activity in A549 cells.
- The highest GL67 lipid ratio (100% GL67) showed the greatest cellular uptake.
Conclusions:
- GL67-based lipoplex nanocarriers are effective for siRNA delivery, showing high internalization efficiency.
- These nanocarriers possess a favorable safety profile, indicating potential for treating genetic diseases.
- The developed system addresses the limitations of naked siRNA, paving the way for advanced genetic therapies.
