Blood contains a wealth of biomarkers that can be used to diagnose cancer at early stages and to successfully manage its treatment. Biomarkers include circulating tumor cells (CTCs) and cell-free DNA (cfDNA), however biomarker isolation and molecular characterization has proven to be challenging due to low abundance and rapid degradation. In contrast, extracellular vesicles (EVs), which also originate from a developing tumor are abundant in blood and other biological fluids, and EV contents (DNAs, RNAs, metabolites, proteins) are stably protected from degradation from the EV lipid bilayer. The 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 individualized, patient-specific care and treatment. BioFluidica, Inc. has demonstrated a platform with superior performance capabilities for the isolation of CTCs using microfluidic technology. Building on this technology, the objective of this Phase I proposal is to develop a novel microfluidic affinity purification (MAP) device and operational peripherals for the automated isolation of disease-associated EVs for downstream molecular profiling. Leveraging our previous success, we will demonstrate improved affinity capture of EVs through advanced microfluidic biochip design (Aim 1). The microfluidic module will be made from polycarbonate that can be molded, allowing for high-scale production at low cost. The chip will be designed to accommodate pipette tips associated with an FDA-compliant liquid handling station to automate operation of the MAP device (Aim 2). We 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 Center (Aim 3). The final product will enrich disease-associated EVs with high recovery and reproducibility, provide high specificity and throughput, and be an automated, cost-effective, and thus commercially viable technology.
Project Terms: Academic Medical Centers; Affinity; Affinity Chromatography; base; Beds; biobank; biochip; Biological; Biological Assay; Biological Markers; Blood; Body Fluids; Breast Cancer Patient; Caliber; cancer cell; cancer diagnosis; cancer type; Cancerous; Carcinoma; Caring; Cell Communication; cell free DNA; Cell Line; Cell physiology; Cells; Cleaved cell; Clinical; Clinical Research; commercially viable technology; cost; cost effective; Culture Media; design; Detection; Development; Devices; Diagnosis; Diagnostic; Diamond; Disease; disease diagnosis; Distant; exosome; experimental study; extracellular vesicles; fibroblast activation protein alpha; genetic analysis; Growth; improved; Injections; Kansas; Lipid Bilayers; liquid biopsy; Liquid substance; malignant breast neoplasm; Malignant Neoplasms; Manuals; Mediator of activation protein; Mesenchymal; Messenger RNA; Methods; Microfluidic Microchips; microfluidic technology; Microfluidics; minimally invasive; Modality; Molds; Molecular; Molecular Analysis; Molecular Profiling; Monitor; Monoclonal Antibodies; Monte Carlo Method; nanoparticle; Neoplasm Circulating Cells; Neoplasm Metastasis; neoplastic cell; new technology; novel; Nucleic Acids; Oligonucleotides; operation; Patients; Performance; Peripheral; personalized approach; Phase; phase 2 study; polycarbonate; Population; precision medicine; Primary Neoplasm; Production; prognostic; protein metabolite; Proteins; prototype; Pump; Recording of previous events; Recovery; Reproducibility; Robotics; Sampling; Secure; simulation; SKBR3; Source; Specificity; success; Syringes; System; TACSTD1 gene; Technology; Testing; Therapeutic; Time; tool development; Tube; tumor; tumor progression; Uracil; Vesicle;
