Blood contains a wealth of biomarkers that can be potentially used to diagnose cancer at early stages and to successfully manage its treatment. Biomarkers include circulating tumor cells (CTCs), cell-free DNA (cfDNA) and extracellular vesicles (EVs), which comprise the âliquid biopsyâ portfolio, originate from the developing primary tumor and the metastatic sites. However, the disease-associated blood-based biomarkers are rare, and the isolation of each from a âliquid biopsyâ requires different approaches. Thus, the development of new technologies that afford efficient and cost-effective isolation of each biomarker holds the key to advancing precision medicine approaches for individualized, patient-specific care, and treatment. BioFluidica has demonstrated a platform with superior performance capabilities of the isolation of CTCs using microfluidic technology. Building on this technology, the objective of this Phase I proposal is to develop a novel microfluidic platform and operational peripherals for the automated, solid phase extraction (SPE) of cfDNA. Leveraging our previous success, we will develop a SPE microfluidic chip for the efficient isolation of cell free DNA fragments from plasma (Aim 1). The microfluidic chip 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 chip (Aim 2). We will demonstrate the utility of our microfluidic chip by enriching cfDNA isolated from plasma spiked with cfDNA containing mutant alleles of several oncogenes ranging from 0.1-5%. Enriched cfDNA will be analyzed by a highly sensitive ligase detection reaction. The final SPE product will enrich cfDNA with high recovery and reproducibility, provide high specificity and high-throughput, and be an automated, cost-effective, and commercially viable technology.
Public Health Relevance Statement: Blood-based liquid biopsies are emerging as the preferred approach to manage a variety of cancer-related diseases, all from a simple blood draw. In this proposal, we aim to develop a cost-effective and highly efficient approach for the isolation of cell free DNA (cfDNA) using an automatic microfluidic approach. This technology will advance a patient-centric, precision medicine approach to diagnose and improve treatment success for patients with a variety of cancer-related diseases.
Project Terms: Alleles; Area; base; bed capacity; Biological Assay; Biological Markers; Biopsy; Blood; Blood donor; Blood specimen; blood-based biomarker; Buffers; cancer diagnosis; Cancer Patient; Caring; cell free DNA; Cell Separation; Cells; Clinical; Clinical Research; commercially viable technology; Complement; Copy Number Polymorphism; cost; cost effective; design; Detection; Development; Devices; Diagnosis; Diagnostic; Dideoxy Chain Termination DNA Sequencing; Discrimination; Disease; Disease Progression; DNA; Ensure; extracellular vesicles; feeding; fluid flow; Frequencies; Genetic Polymorphism; Genomic DNA; Immobilization; improved; individualized medicine; industry partner; Injections; Length; Leukocytes; Ligase; liquid biopsy; Liquid substance; Malignant Neoplasms; Methods; Methylation; Microfluidic Microchips; microfluidic technology; Microfluidics; Molds; Molecular Analysis; Molecular Profiling; Monitor; multidisciplinary; mutant; Mutation; Mutation Detection; Neoplasm Circulating Cells; neoplastic cell; new technology; Normal Cell; novel; Nucleotides; Oncogenes; operation; Operative Surgical Procedures; Pathology; Patient Care; Patients; Performance; Peripheral; personalized approach; personalized medicine; Phase; Plant Roots; Plasma; polycarbonate; Population Sizes; precision medicine; Primary Neoplasm; Production; Protocols documentation; Pump; Reaction; Recording of previous events; Recovery; Reference Standards; Reporting; Reproducibility; Robotics; Secure; Series; Site; Solid; Specificity; success; Syringes; Systems Analysis; Technology; Time; Treatment Protocols; tumor; Tumor Markers; Tumor-Derive
