CD44 targeted delivery of siRNA by using HA-decorated nanotechnologies for KRAS silencing in cancer treatment

Area of Science:

• Biotechnology and Nanomedicine
• Molecular Biology and Genetics
• Oncology

Background:

• KRAS mutations drive tumor growth in various cancers, including colorectal cancer.
• Targeting KRAS remains challenging due to the difficulty in developing effective small molecule inhibitors.
• Gene silencing using small interfering RNA (siRNA) offers an alternative therapeutic strategy for KRAS-driven cancers.

Purpose of the Study:

• To develop targeted ternary nanocomplexes for delivering anti-KRAS siRNA to colorectal cancer cells.
• To leverage the hyaluronic acid (HA)-CD44 interaction for selective targeting of CD44-positive cancer cells.
• To evaluate the efficacy of siRNA delivery and KRAS gene silencing in a relevant cancer model.

Main Methods:

• Ternary nanocomplexes were formulated using siRNA, polycations (poly(hexamethylene biguanide) or chitosan), and hyaluronic acid (HA) coating.
• Nanoparticle characterization included assessment of size, surface charge, and siRNA protection.
• In vitro studies involved flow cytometry for nanoparticle uptake, confocal microscopy for siRNA release, and RT-qPCR for KRAS gene silencing in HCT-116 cells.

Main Results:

• Characterization confirmed the physico-chemical properties and stability of the HA-coated nanocomplexes.
• Demonstrated efficient nanoparticle internalization and siRNA release into the cytosol of CD44+/KRAS+ colorectal cancer cells.
• Achieved significant KRAS gene silencing in HCT-116 cells, confirming the therapeutic potential.

Conclusions:

• HA-decorated nanocomplexes provide a targeted delivery system for anti-KRAS siRNA in colorectal cancer.
• This approach effectively silences KRAS in cancer cells and shows enhanced uptake when co-cultured with fibroblasts.
• The study highlights the potential of targeted gene silencing via nanocomplexes as a therapeutic strategy for KRAS-mutated cancers.