ROS Sensitivity-Programmed DNA Origami Sensors for Point-of-Care Urinalysis

Excessive generation of reactive oxygen species (ROS) in the liver is a critical molecule event in the early stages of acute liver injury (ALI), preceding hepatocyte necrosis and the leakage of alanine aminotransferase (ALT). Urinalysis of liver ROS holds significant potential for point-of-care diagnostics of early-stage ALI, yet remains highly challenging as the liver ROS is difficult to metabolize into urine reliably. To address this challenge, we herein engineered DNA origami nanostructures (DONs) with varying nick numbers, and established a positive correlation between nick number and the sensitivity to ROS-induced degradation of DONs. Building on this premise, we developed ROS sensitivity-programmed DNA origami sensors (DOSs) by orthogonally assembling targeting and signal modules with DONs. DOSs can specifically accumulate in the liver and undergo ROS-triggered intact architecture-to-degradation debris pharmacokinetic conversion, thus transforming the liver ROS into measurable synthetic urinary markers. Notably, this transformation efficiency positively depended on nick number of DOSs, enabling point-of-care urinalysis of the onset of different levels of ALI at least 4 h earlier than the ALT-based clinical blood test, with a maximum area under the curve of 0.94. We anticipate that this strategy may be extended for early diagnosis of various ROS-related diseases via a simple urine test.