The maintain the transparency of the eye lens, it depends on the native tertiary structures and solubility of the lens crystallin proteins over a lifetime. Therefore, understanding the process of in vitro/in vivo stability of chaperones small molecules or gene core-loaded with carriers and their biodistribution in the cellular lens uptake is critical for developing effective delivery system of cataract treatment. Under our previously observed that polymeric micelles (PM) can encapsulate a Forster resonance energy transfer (FRET)-pairs or RhB-DNA and retain their integrity complexes within the cornea region two hour after administration. In addition, these PM could distribute among the cornea, lens capsule area as well as through lens cortex and nucleus. Therefore, our major objective of this project is to design a lens nano-sized particle of labeled FITC-PM with chaperones (PCs), lanosterol related compound or crystallins protein genes or autophagy gene, to achieve a timed and lens tissue distribution for observation of chaperone or gene with PM transfer. The optimization of physicochemical kinetic formation of PM (size/shape), in vitro and in vivo stability/retention, distribution of tissue/cells pharmacokinetics of lanosterol and function expression properties of crystalline protein genes in lens region will be systematically investigated. In year 1, we will synthesize fluorochrome-labeled polymer (PM), FITC-PM and core loaded rhodamine-lanosterol or gene for in vitro FRET observation. A time-resolved fluorescence measurement of a donor-acceptor pair for lanosterol or gene core loaded with PM will be evaluated as well as release/ stability of gene from PM. In year 2, we will focus on the assessment of the penetration and distribution of lanosterol or crystalline plasmids delivered with FITC-PM. The permeation orientation/distance of rhodamine-genes complexes with FITC-PM will be measured by a donor-acceptor pair of FRET. Pre-treated with different permeation inhibitors (caveolae-, clathrin-dependent endocytosis, energy dependence etc.) will be evaluated the influence of penetration distance/distribution pathway between both of labeled-genes and PM. In year 3, we will apply an eye drop rhodamine-lanosterol or lens crystalliln or autophagy plasmids/FITC-PM delivery at cataract lens region in vivo. The pharmacokinetic biodistribution lanosterol or crystalline genes with FITC-PM in induction of cataract by sodium selenite in lens animal model will be examined by multi-modality image, real time confocal microscopy, electron microscopy, western blotting, and immunohistochemistry observations. The transport/permeability rate will be also calculated to evaluate the modulation of barrier rates of plasmid with PM's permeability.
|Effective start/end date||8/1/18 → 7/1/19|
- eye drop
- polymeric micelles