Abstract
Vitamin C is an essential micronutrient with biological functions that extend beyond its antioxidant properties, notably serving as a cofactor for iron- and 2-oxoglutarate-dependent dioxygenases, including the ten-eleven translocation (TET) family of DNA demethylases. Loss-of-function mutations in TET2 are frequent drivers of hematological malignancies, and recent studies have shown that high-dose vitamin C administered orally or parenterally can restore TET2 enzymatic activity in circulating white blood cells and suppress leukemogenesis. However, the intracellular pharmacokinetics and hematopoietic tissue distribution of vitamin C, which are critical to its therapeutic efficacy, remain poorly characterized. To address this, we synthesized a bioorthogonal, “clickable” vitamin C analog that enables precise visualization through strain-promoted azide-alkyne cycloaddition (SPAAC) using flow cytometry and fluorescence microscopy. This innovative tool allows high-resolution tracking of vitamin C uptake and subcellular localization in vitro and in vivo. Using this approach, we quantified vitamin C distribution across diverse hematopoietic lineages and in murine models of myeloid leukemia under varied dosing and delivery conditions. Our findings reveal cell-type–specific uptake patterns that underscore vitamin C’s potential as a targeted epigenetic therapeutic in hematologic malignancy. This study establishes a novel framework for investigating cellular-level vitamin C bioavailability and advances the rational development of vitamin C-based epigenetic therapies for leukemia.