Tracking Selective Internalization and Intracellular Dynamics of Modified Chitosan Polymeric Micelles of Interest in Primary Hyperoxaluria Diseases.

Primary hyperoxalurias (PHs) represent rare diseases associated with disruptions in glyoxylate metabolism within hepatocytes. Impaired glyoxylate detoxification in PH patients results in its accumulation and subsequent conversion into oxalate, a process catalyzed by the hepatic lactate dehydrogenase A enzyme (LDHA). Targeting this enzyme selectively in the liver using small organic molecules emerges as a potential therapeutic strategy for PH. However, achieving selective hepatic inhibition of LDHA poses challenges, requiring precise delivery of potential inhibitors into hepatocytes to mitigate adverse effects in other tissues. Our recent efforts focused on the design of polymeric micelle nanocarriers tailored for the selective transport and release of LDHA inhibitors into liver tissues. In this study, we synthesized and assessed the internalization and disaggregation dynamics of chitosan-based polymeric micelles in both hepatic and nonhepatic cell models using live-cell imaging. Our findings indicate that lactonolactone residues confer internalization capacity to the micelles upon exposure to cells. Moreover, we demonstrated the intracellular disaggregation capacity of these nanocarriers facilitated by the cystamine redox-sensitive linker attached to the polymer. Importantly, no cytotoxic effects were observed throughout the experimental time frame. Finally, our results underscore the higher selectivity of these nanocarriers for hepatic HepG2 cells compared to other nonhepatic cell models.