The limited supply of red blood cells (RBC) and platelets is a serious medical issue that can have life-threatening consequences for transfusion-dependent patients, particularly those who develop platelet allo-immunity. The low prevalence of O Rh-negative 'universal donor'blood type in the general population (<8% in Western countries and <0.3% in Asia) further intensifies the consequences of blood shortages for emergency situations where blood supply and/or typing is limited. Based on the pioneering work of Yamanaka and Thomson, human induced pluripotent stem cells (hiPSCs) offer the possibility to generate patient-specific stem cells without destruction of embryos. However, current iPSCs suffer from major drawbacks including multiple viral integrations and remaining transgenes at various chromosomal locations, any of which may cause unpredictable genetic dysfunction and/or tumor formation (Yamanaka, 2009). In this proposal, based on our recent """"""""proof-of-concept"""""""" results, we hypothesize that hiPS cells suitable for clinical translation can be generated by direct delivery of reprogramming proteins attached to a cell penetrating peptide. In particular, we will generate fully reprogrammed hiPS cell lines from both healthy and O(-) subjects by direct protein delivery and test whether these cells can be propagated and expanded in vitro indefinitely, thus providing a potentially inexhaustible and donor-less source of blood lineage cells. This proposal will fully optimize the protein reprogramming methods to establish a highly efficient and safe way of reprogramming human tissues without genetic manipulation, and will address whether these hiPS cells can be used as a personalized platelet source and/or universal RBC source.
Although still in its infancy, """"""""induced pluripotent stem cell (iPSC)"""""""" technology has the potential to revolutionize biomedical research, disease mechanism studies, and customized cell-based therapies. To explore the potential of iPSC's as a source of universal red blood cells and personalized platelets, we propose to establish and characterize clinically viable iPSC lines from healthy O Rh-negative [O(-)] subjects by direct delivery of reprogramming proteins without the use of viruses or foreign-DNA vectors. Using these protein-induced iPS cells, we will address whether they can be propagated and expanded in vitro indefinitely into blood cell progenitor's lineages that can be used as a donor-less source of universal red blood cells and/or personalized platelets.
|Feng, Qiang; Shabrani, Namrata; Thon, Jonathan N et al. (2014) Scalable generation of universal platelets from human induced pluripotent stem cells. Stem Cell Reports 3:817-31|
|Park, Hansoo; Kim, Dohoon; Kim, Chun-Hyung et al. (2014) Increased genomic integrity of an improved protein-based mouse induced pluripotent stem cell method compared with current viral-induced strategies. Stem Cells Transl Med 3:599-609|
|Park, Kyung-Soon; Cha, Young; Kim, Chun-Hyung et al. (2013) Transcription elongation factor Tcea3 regulates the pluripotent differentiation potential of mouse embryonic stem cells via the Lefty1-Nodal-Smad2 pathway. Stem Cells 31:282-92|