The groundbreaking discovery of induced pluripotent stem cells (iPSCs) has opened a new page for developing personalized cell-based therapies against various devastating human diseases without ethical concerns of using human embryonic stem cells. However, the future clinical applications of iPSCs remain in doubt due to potential risks of undesired genomic alteration and tumor formation. Generation of iPSCs by transient expression of pluripotent factors has decreased the safety concerns yet the reprogramming efficiency remains low. In this application, we propose to develop an improved, cell-permeable protein-based method to efficiently generate iPSCs. We have developed a rational approach to systematically engineer, produce, and validate robust, cell-penetrating reprogramming peptides that will overcome the key barriers to achieve high-efficiency and safe cellular reprogramming. The technology will enable us to efficiently produce patient-compatible iPSCs with the best safety profiles so as to derive other cell types safely for transplantation or for other types of clinical applications. Two Aims are proposed: Specific Aim 1 (Months 1-6): To construct a library of cell permeable human iPSC reprogramming proteins with modifications that would increase their reprogramming efficiency at Vivoscript Inc. Specific Aim 2 (Months 7-12): To evaluate the efficiency of these proteins in iPSC generation in Chen's lab at Sanford-Burnham Medical Research Institute.
Public Health Relevance Statement: Public Health Relevance: We are developing robust, cell-penetrating peptides to overcome the limitations of current technologies for an efficient generation of human induced pluripotent stem cells (iPSCs). Our novel technology will enable us to efficiently produce patient-compatible iPSCs with the best safety profiles so as to benefit the public by deriving other cell types from these iPSCs for future clinical therapy or drug screens.
Project Terms: Adult; Adverse effects; Animal Model; Applications Grants; Back; base; Cell Nucleus; Cell Therapy; cell type; Cells; Clinical; clinical application; Culture Media; Cytoplasm; cytotoxicity; design; DNA Sequence Alteration; Elements; Endosomes; Engineering; Environment; Ethics; Fibroblasts; Future; Gene Expression; Generations; Genetic; Genome; Genomics; Green Fluorescent Proteins; Human; human disease; human embryonic stem cell; Image; Immune response; improved; in vitro Model; induced pluripotent stem cell; interest; Libraries; Life; Lipids; Medical Research; Messenger RNA; Methods; Modification; Monitor; MyoD Protein; new technology; Nuclear; Nuclear Export; Nuclear Import; Nuclear Translocation; Outcome; Patients; Peptides; Preclinical Drug Evaluation; Proteins; public health relevance; Pump; Reproducibility; Research Institute; Risk; Safety; Skeletal muscle structure; Somatic Cell; stem; Stem cells; System; Techniques; technological innovation; Technology; Testing; Therapeutic; Therapeutic Studies; Time; transcription factor; Transplantation; tumor; Uncertainty; Validation; Viral Vector
