EpCAM aptamer-engineered, azadiradione-loaded, chemo-photo-responsive nano-erythroliposomes for the treatment of triple-negative breast cancer

Abstract

This research aimed to engineer azadiradione (AZ) and IR780-enabled, epithelial cellular adhesion molecule (EpCAM) aptamer (Apt) grafted, targeted nano-erythroliposomes (Apt/Eryt/AZ/IR780) for chemo-photo combined treatment of triple-negative breast cancer (TNBC). Physicochemical characterization revealed average particle size and polydispersity index (PDI) of 106.8±3.76 nm and 0.176 ±0.03, respectively, and zeta potential of 28.75±0.65 mV. Compared to formulations that were not NIR triggered, ∼85 % of the AZ was released in 8 h from NIR-exposed formulation. EpCAM expression analysis unequivocally demonstrated that EpCAM levels were significantly elevated in 4T1 cells compared to MDA-MB-231 cells. Cellular uptake analysis revealed strong fluorescence intensity with Apt/Eryt/AZ/IR780 with NIR exposure compared to non-aptamer formulations. Dose-dependent MTT assay (100 mM, 150 mM, and 200 mM concentration of AZ) revealed that <20 % of cells survived after treatment with Apt/Eryt/AZ/IR780 and NIR exposure. The cell viability results were further validated by employing reactive oxygen species (ROS) generation determination, mitochondrial membrane potential (MMP) inhibition, and Caspases-8 assay. In particular, 4T1 cells were found to be sensitive to AZ by decreasing MMP activity (∼17-fold reduction compared to control) and increasing Caspase-3 activity (∼544-fold enhancement compared to control). A hemocompatibility assay revealed a non-significant interaction (<0.9 %) of Apt/Eryt/AZ/IR780 with blood components. In addition, stability investigation for 30 days at 30±2 °C and 5±3 °C of Apt/Eryt/AZ/IR780. The findings revealed no significant change in average size (<8.5 %), zeta potential (<25 %), PDI (<20 %), and % of IR780 (<14 %) and AZ (<13 %) at 5±3 °C in 90 days.