Enhanced cytoplasmic delivery of therapeutics by NIR and graphene oxide-mediated thermally-induced endosome/lysosome leakage

ABSTRACT: Numerous clever and creative and approaches to develop non-viral delivery vehicles have been described and every year brings additional progress. However, a bottleneck to nearly every strategy is endosomal/lysosomal vesicle escape. Transfection frequencies under controlled conditions with in vitro models rarely exceed 50-70% efficiency in the best model systems and are often lower, depending on cell type and cargo. In this proposal we will test a new way to improve vesicle escape that can be applied to almost any existing delivery technology. An important parameter in lipid bilayer membrane integrity is temperature. As the temperature rises, the kinetic energy imparted to the membrane lipids increases the possibility for local disorganization and rupture of the bilayer. We propose to test the hypothesis that controlled heating of endosomal/lysosomal vesicles containing a variety of delivery vehicles and cargo will enhance membrane breakage and improve delivery efficiency. Heating will be accomplished by exploiting the ability of nanoparticles to absorb near infrared (NIR) light and convert it to heat. The use of NIR has the advantage that tissue is relatively transparent in this region of the spectrum and that vesicle disruption can be targeted to tissue and organs where the laser is aimed. BENEFIT: The combination of a delivery vehicle that can efficiently cross the cell membrane and has high near infrared absorption enables the timed and on-demand transport and release of therapeutic agents. Such benefit will allow for quick response to the chemical and biological threats to which those on the battlefield may be exposed. Therapeutic agents that are difficult to administer or require high doses to achieve high efficacy may be more efficiently used when coupled to NIR-active nanoparticles which can transcend the cell membrane and enter intracellular vesicles and, subsequently release their payload upon NIR activation. Finally, novel drug combinations may be administered to patients that have acquired recurrent or resistance disease where existing therapeutic regimens have failed.