Circulating Red Blood Cell Based Nanosensors for Continuous, Real-Time Drug Monitoring

ABSTRACTNanosensor technology for continuous monitoring of proteins in vivo would enable researchers to track thedynamics of biomolecule expression as it pertains to disease pathogenesis or predicting therapeutic efficacy,with results available in real-time. An example where this diagnostic ability would be groundbreaking is in thecontext of understanding cytokine release syndrome (CRS). CRS is a systemic inflammatory response thatarises when the immune system is overstimulated, leading to extreme toxic events such as multiple organdysfunction1. There is now increasing evidence to suggest that the development of severe cases of COVID-19can be attributed to onset of CRS. It has been revealed that serum levels of cytokines like IFNγ, IL-6, sIL-2Rα,and IL-10 can be significantly elevated in patients with severe CRS, before the apparent onset of severesymptoms. However, the use of cytokines as biomarkers of CRS would require a rapid, minimally invasivediagnostic assay, which is currently unavailable, slowing animal studies of COVID-19/CRS. Recently publishedresearch from the Clark laboratory has demonstrated a proof-of-concept DNA-based sensor for minimallyinvasive detection of IFNγ, one of the cytokines that has been proposed as a biomarker for predicting thepotential for onset of severe CRS. This design was inspired by advances in DNA nanotechnology, which enableresearchers to create functional nanostructures with site-specific modifications based on the complementarybase-pairing rules of DNA. The open or closed state of the sensor could be determined through differentialsignals as detected with optical imaging. Drawing from recent advances in DNA origami design and stabilizationtechnology, we hypothesize that we can improve on this work and produce a robust platform for opticalmonitoring of IFNγ in real-time by (1) enhancing the rigidity of our DNA platform and (2) deploying protectionstrategies to ionically stabilize the construct in biological solutions. This project aims to advance current analyticalstrategies for immunological diagnostics by providing researchers with a powerful tool to probe biomoleculedynamics toward in vivo use with existing optical imaging platforms. The one-year project will result in a robusttool developed for animal research. The goal will be to commercialize and distribute the sensor for COVID-19studies, as well as other immune system research.