Developing efficient and safe cell-permeable reprogramming peptides for generation of iPS cells.

描述（由申请人提供）：诱导多能干细胞（iPSC）的突破性发现为开发针对各种毁灭性人类疾病的个性化细胞疗法打开了新的一页，而无需考虑使用人类胚胎干细胞的伦理问题。然而，由于不期望的基因组改变和肿瘤形成的潜在风险，iPSCs的未来临床应用仍然存在疑问。通过多能因子的瞬时表达产生iPSC减少了安全性问题，但重编程效率仍然很低。在本申请中，我们提出开发一种改进的、基于细胞可渗透蛋白质的方法来有效地产生iPSC。我们已经开发了一种合理的方法来系统地设计，生产和验证强大的细胞穿透重编程肽，这将克服实现高效和安全的细胞重编程的关键障碍。该技术将使我们能够有效地生产具有最佳安全性的患者相容性iPSC，以便安全地衍生其他细胞类型用于移植或其他类型的临床应用。提出了两个目标：具体目标1（第1-6个月）：在Vivoscrip Inc.构建具有将增加其重编程效率的修饰的细胞可渗透的人iPSC重编程蛋白的文库。具体目标2（第7-12个月）：在Sanford-Burnham医学研究所的Chen实验室中评估这些蛋白质在iPSC生成中的效率。

DESCRIPTION (provided by applicant): 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.