Parallel and high throughput isolation of cancer cell derived extracellular vesicles

Blood contains a wealth of biomarkers that can be used to diagnose cancer at early stages and to successfully manage its treatmentBiomarkers include circulating tumor cellsCTCsand cell free DNAcfDNAhowever biomarker isolation and molecular characterization has proven to be challenging due to low abundance and rapid degradationIn contrastextracellular vesiclesEVswhich also originate from a developing tumor are abundant in blood and other biological fluidsand EV contentsDNAsRNAsmetabolitesproteinsare stably protected from degradation from the EV lipid bilayerThe development of new technologies that afford the efficient and cost effective isolation of purified EVs populations holds the key to advancing precision medicine approaches for individualizedpatient specific care and treatmentBioFluidicaInchas demonstrated a platform with superior performance capabilities for the isolation of CTCs using microfluidic technologyBuilding on this technologythe objective of this Phase I proposal is to develop a novel microfluidic affinity purificationMAPdevice and operational peripherals for the automated isolation of disease associated EVs for downstream molecular profilingLeveraging our previous successwe will demonstrate improved affinity capture of EVs through advanced microfluidic biochip designAimThe microfluidic module will be made from polycarbonate that can be moldedallowing for high scale production at low costThe chip will be designed to accommodate pipette tips associated with an FDA compliant liquid handling station to automate operation of the MAP deviceAimWe will demonstrate the utility of the microfluidic device to distinguish between cancerous and non cancerous biological samples obtained from the biorepository at the Kansas University Medical CenterAimThe final product will enrich disease associated EVs with high recovery and reproducibilityprovide high specificity and throughputand be an automatedcost effectiveand thus commercially viable technology Blood based liquid biopsies are emerging as the preferred approach to manage a variety of cancer related diseasesall from a simple blood drawIn this proposalour objective is to develop a cost effective and highly efficient approach for the isolation of extracellular vesicles using an automatic microfluidic approachThis technology will advance a patient centricprecision medicine approach to diagnosing and improving treatment success for patients with cancer