pRNA Nanoparticles as Tools to Improve Human Pluripotent Stem Cell-based Transplantation Strategies for Retinal Degenerative Disease

This project combines pRNA nanotechnology with human stem cell biology for the purpose of improving retinal cell transplantation strategies for atrophic AMD. We propose to use three-way junction pRNA (3WJpRNA), a nanoparticle with great therapeutic potential, as an innovative tool to temporarily label, sort, and improve viability of neuroretinal cells derived from human pluripotent stem cells (hPSCs). 3WJpRNA can be manufactured to carry three functional moieties, including fluorophores, small molecule inhibitors, and receptor ligands for cell-specific targeting. In collaboration with fellow BIMR member, Dr. Peixuan Guo (on as consultant), Dr. Shu and her TBN graduate student will test currently available fluorophore-conjugated 3WJpRNAs that target cells expressing folate or EGFR receptors, hypothesizing that these 3WJpRNAs will transiently label subpopulations of neuroretinal cell types within our hPSC optic vesicle cultures. In addition, Dr. Shu and her student will create a modified version of each of these 3WJpRNAs that includes an inhibitor of p160Rho-associated kinase (Y27632). The tracking and pro-survival properties of Y27632-containing 3WJpRNAs will be evaluated in hPSC-derived neuroretinal cells following FAC-sorting and transplantation into the P23H rat model. Overall, this project is designed to address a significant and unsolved problem facing stem cell transplantation: the need to enrich for and improve survival of selected neuroretinal progeny using a nontoxic, nonimmunogenic strategy that does not require permanent genetic modification. Although this application will be initially tested in a retinitis pigmentosa model, our findings will be directly relevant for all retinal degenerative diseases, including atrophic macular degeneration.