NHMRC Early Career Fellowship (CJ Martin - Overseas Biomedical Fellowship): Early detection of disease using fluorescence resonance energy transfer (FRET) diagnostics (2014–2018)
There has recently been a large research focus on pre-clinical, non-invasive technologies that can effectively detect and diagnose diseases at very early stages. This project aims to develop an understanding of the use of switchable polymeric nanoparticles for early detection of these diseases.
This proposal will initially focus on developing a platform technology that will correlate the physico-chemical properties of a degradable polymer with Fluorescence Resonance Energy Transfer (FRET) imaging. Variation in synthetic parameters along with a well-known biological test bed will enable us to establish a methodology for investigating key mechanisms of polymer nanoparticle internalization into cells. Incorporation of slowly degrading monomers into the polymer structure will enable larger, harder to excrete particles to be removed from the body once internalized.
The second part of this proposal will focus on incorporating specific targeting ligands and sequences for efficient cell uptake. Folic acid and peptide aptamers will be used to target melanoma cells, and variation in the presentation of these targeting modalities to receptor proteins will enable us to observe how the spatial configuration of polymer nanoparticles affects efficacy of receptor-mediated uptake.
Finally, this proposal will focus on obtaining fundamental information of cell uptake in vitro that can be translated to in vivo for these devices. Investigation into cellular uptake of these particles will be conducted using a test-bed of known melanoma cell lines using FRET and high resolution confocal microscopy, followed by in vivo studies to verify the targeting efficacy of the molecules and monitor internalization using fluorescence imaging. This proposal will contribute significantly to the progress of this field through advances in both understanding cellular delivery mechanisms and stimuli specific imaging.