GrantR33CA
Molecular Beacon Based Extracellular mRNA and Protein Detection for Early Cancer Diagnosis
- Activity
- R33
- Institute
- CA
- Fiscal years
- 2017–2017
- Amount counted
- $1,588,359
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Abstract public domain (NIH RePORTER)
Abstract Many cancer types including pancreatic cancer have very high mortality rates because it is difficult to detect their presence in the early stage. Current biomarkers for pancreatic cancer such as serum CA-19-9 protein face high false positive and negative rates. The proposed studies seek to develop a robust and user-friendly novel molecular beacon and lipoplex nanoparticle biochip technology and validate its clinic potential by detecting Glypican 1 mRNA and protein in extracellular vesicles from pancreatic cancer patient serum samples for imminent use in the field of early cancer diagnosis. The specific aims of this project are: Aim 1: Development of Robust Biochip Technology. 1-1. Improved MBs for high stability and mis-match specificity; 1-2. Standard vesicles (SVs) for chip and analyzer calibration and analytical quantification; 1-3. Non-TIRF fluorescence microscope as a low-cost analyzer and automated software module for fast and user-friendly image acquisition and analysis. Specific Aim 2: Multi-site Technology Validation for Pancreatic Cancer Diagnosis. 2-1. Comparison with qRT-PCR, NGS and ELISA for performance, cost, assay time, and user friendliness. 2-2. Multi-site blind validation: Lab-to-lab repeatability and blind validation comparison at OSU and MSKCC using patient and normal donor serum samples.
Public health relevance public domain (NIH RePORTER)
Project Narrative Current biomarkers for pancreatic cancer such as serum CA-19-9 protein face high false positive and negative rates. We present a novel technology that enables the direct analysis of biomarkers within individual extracellular vesicles. We accomplish this without the need for mixing together the contents of all extracellular vesicles or amplification, which results in a faster assay with more sensitive results. Our novel technique utilizes a tethered lipoplex nanoparticle biochip containing molecular beacons to directly capture individual extracellular vesicles and detect biomarkers within them. This amplification-free analysis technique is faster, less expensive, and less prone to error than the current q-PCR technology, and could be a valuable tool in the emerging area of liquid biopsy.