A Protocol To Characterize Peptide-Based Drug Delivery Systems for miRNAs

Area of Science:

• Biochemistry
• Nanotechnology
• Pharmacology

Background:

• Micro RNAs (miRNAs) are key regulators of metabolic processes, but naked miRNAs have poor stability and cellular uptake.
• Altered miRNA expression is linked to metabolic and genetic diseases, necessitating effective drug delivery systems (DDS).

Purpose of the Study:

• To develop and characterize a novel peptide-based drug delivery system (DDS) for miRNA-27a.
• To evaluate the physicochemical properties of the DDS for potential miRNA replacement therapy in metabolic disorders.

Main Methods:

• Dynamic light scattering (DLS) and electrophoretic light scattering were used to assess DDS size and charge.
• Atomic force microscopy (AFM) was employed to investigate DDS shape, stability, and substrate adhesion.
• Physicochemical properties were analyzed to understand their impact on cellular uptake.

Main Results:

• The DDS, composed of miRNA-27a and N-TER peptide, exhibited agglomeration despite expected surface charge stabilization.
• AFM confirmed DDS agglomeration and revealed adhesion to both hydrophilic and hydrophobic surfaces due to the peptide's amphipathic nature.
• Physicochemical properties were identified as critical for cell internalization and successful DDS development.

Conclusions:

• The N-TER peptide-based DDS demonstrates unique adhesion properties relevant for miRNA delivery.
• Understanding DDS physicochemical characteristics is crucial for developing effective peptide-based miRNA replacement therapies for metabolic diseases like obesity.