Intrinsic Anti-Angiogenic Activity of siRNA - Supplemental - Equipment

The parent proposal was to test the hypothesis that both targeted and non-targeted siRNAs activate TLR3 by direct interaction and trigger a cellular program of vascular growth suppression. The specific aims of the proposal were to further define the interaction between s1RNA and TLR3 and to determine the precise anti-angiogenic mechanisms of siRNA-induced CNV suppression in mice. In this supplement proposal for infrastructure support, we seek to acquire a state-of-the-art analytic research tool that will significantly enhance our scientific approach to the proposed experimental aims in R01-EY018836. The requested equipment is a specialized machine capable of performing in situ mass spectrometry (MS) on almost any sample format including tissue sections. The technology is abbreviated MALDI-TOF, and its application to the biomedical sciences is just beginning to demonstrate an unparalleled power for the direct quantification and localization of any biologic molecule of interest, drug, metabolite, and/or toxic byproduct. We plan to incorporate MALDI-TOF imaging analyses in order to achieve the highest spatial and spectral resolution possible for monitoring and profiling cytokine kinetic investigations detailed in R01-EY018836. There are over 100 experimental designs detailed in the parent proposal that will benefit from this updated infrastructure. While the laboratory has made solid progress utilizing conventional molecular biological techniques towards the Specific Aims set forth in the parent proposal, the acquisition of this ultra-modern tool will accelerate our research capabilities and lead to further discovery within the time constraints of the funding period. Furthermore, my laboratory's expertise in age-related macular degeneration combined with the collaborative commitment of Bert Lynn PhD, Director of Mass Spectrometry at University of Kentucky, will create one of the few, and perhaps only, research groups in the world applying this next-generation instrument to investigations of the molecular mechanisms that cause this devastating disease.