PROJECT SUMMARY Nucleic acid nanoparticles (NANPs) have the ability to simultaneously deliver a cocktail of multiple therapeutic nucleic acids (TNAs) that can treat diseases by targeting several biological pathways. However, in order to be functional, NANPs require a delivery vehicle to enter cells while being protected from naturally occurring nucleases. There have been studies showing that amine-terminated polyamidoamine (-NH2) PAMAM dendrimers have successfully delivered TNAs such as small interfering RNAs, and micro RNAs into cells while efficiently protecting them, but these studies deep dive into only one generation of dendrimers. This proposal aims to investigate and compare five different generations of (-NH2) PAMAM dendrimers to interpret how each will behave as a carrier for several representative NANPs. A second delivery platform that will be investigated are exosomes that are harvested from cancerous and non-cancerous human cell lines. Exosomes naturally pass biological information from cell to cell and we would be utilizing this natural communication machinery to deliver NANPs. Each exosome carries different biomarkers corresponding to the cell line of origin, and these biomarkers can be used to identify the exosome type and tune their biodistribution, immunorecognition and cellular uptake. Along with exosomal identification, the type of the NANP embedded into the exosome complex may additionally contribute to their biological behavior, which is what we would like to investigate in this current proposal. Once competed, this investigation will supply information regarding the role that synthetic and naturally occurring vehicles have on immune response and efficacy, as well as how NANPs’ architectural parameters (e.g., 3D vs 2D vs 1D) influence their delivery and immune response. The broad and long-term objectives for this work would be to categorize a database focused on the in vitro behavior of NANPs complexed to their vehicles to include the immune response, protection, and efficacy of the NANP-carrier platform. In collecting this data, and compiling it, future nucleic acid based therapeutics could be rationally designed with specific parameters and tailored immune recognition and function based on what a patient requires. All proposed experiments will be completed by the applicant who will collaborate with the necessary personal to learn any supplemental techniques needed. In addition to completing the proposed work, the applicant will be serving as a leader in more than one graduate organization where the applicant will be gaining professional, communicative, and leadership skills needed to network when presenting this work at conferences. The University of North Carolina at Charlotte is a space that is experiencing exponential growth that is focused on student success and provides valuable resources for students and researchers to utilize.