Upholding hyaluronic acid's multi-functionality for nucleic acid drug delivery to target EMT in breast cancer

Synthetic nanoparticles can stably encapsulate nucleic acids as active pharmaceutical payloads. Recently, mRNA- and siRNA-based medicines have been successfully approved for preventing or treating infectious or orphan diseases. RNA interference is particularly relevant for cancer therapy, as tumors often involve up- or dys-regulated proteins that drive malignancy. This study aimed to develop a nanoparticulate delivery vehicle that targets EMT-phenotypic breast cancer cells, which lack effective treatment options. These "undruggable" tumors may be addressed by nanoparticles that target EMT-specific cell surface receptors. CD44, a transmembrane protein linked to cancer malignancy and EMT, was identified as a promising candidate. This work investigated the use of hyaluronic acid (HA) in HA-modified polyelectrolyte complexes (polyplexes, Px) for its dual roles as a targeting ligand and a stabilizing stealth-molecule. Various strategies for non-covalently immobilizing HA on the particle surfaces were tested. HAPx nanoparticles demonstrated HA : PEI-ratio dependent stability against competing anionic biomolecules, improved colloidal stability in protein-rich environments mimicking in vivo conditions, and enhanced selectivity and efficacy in targeting E/M-hybrids and EMT-positive cells via CD44-HA mediated endocytosis. Finally, our results indicate different internalization kinetics and efficiencies between CD44v and CD44s isotypes, highlighting the need to consider CD44 heterogeneities in the clinical development of HA-based drug delivery systems.