Automated and Multiplexed Platform for Serum TBI Circulating miRNAs

Traumatic brain injury (TBI) is a complex neurological injury and a major public health problem sustained in both military and civilian populations in the United Sates and worldwide. Current neurological assessment such as the Glasgow Coma Scale and neuroimaging techniques, namely, PET, CT, MRI provide insufficient understanding at a molecular level of the biochemical changes that are ultimately accountable for the observed effects of a TBI. Since genes are sensitive to stress and trauma, epigenetic modifications that involve non-coding RNAs such as circulating microRNAs, have received attention as the possible mechanism for the development of a number of complex indications such as cardiovascular diseases, cancers, and neurological diseases, in particular, TBI. Blood-based miRNA biomarkers are on the verge of a breakthrough for diagnosis, treatment and prognosis of diseases. However, existing miRNA assays such northern blot-based platforms, reverse transcription PCR (RT-PCR), microarray, and next-generation sequencing require extensive preparation involving extraction, reverse transcription of miRNA into cDNA, amplification followed by data analytics. Thus, automated and multiplexed devices that can quantify the levels of circulating miRNAs in biological samples are highly desired. Luna Labs USA, LLCproposes to develop a droplet assay, an automated detection platform designed to assess circulating miRNAs in TBI serum samples. This straightforward, sensitive detection protocol does not require nucleic acid purification, washing steps, or amplification, and is capable to differentiate between exosome-derived and cell-free miRNAs. In Phase I feasibility, Luna LabsĀwill fabricate microfluidic device and integrate onto the assayÆs fluorescence read-out, interrogate multiple miRNAs in both cell-free and exosomal forms, determine assayÆs performance, establish use with readily available clinical serum samples, and establish integration of assay components. In Phase II transition effort, work will initiate extensive multiplexed detection studies and protocol validations, evaluate operation and effectiveness of utilizing an integrated system, and perform complete demonstration from biomatrix to detection of miRNA quantification aiming to meet point-of-care bedside diagnostic tool requirements and FDA compliance.