Hematopoietic stem and progenitor cells derived from hemogenic endothelial cells by episomal plasmid gene transfer

A method of cellular replacement therapy in a subject in need thereof, the method comprising implanting or transplanting to the subject human hematopoietic stem cells (HSCs) or hematopoietic stem and progenitor cells (HSPCs) engineered to exogenously express transcription factors ETS-related gene (ERG), Homeobox protein A9 (HOXA9), Homeobox protein A5 (HOXA5), Ligand Dependent Nuclear Receptor Corepressor (LCOR) and Runt Related Transcription Factor (RUNX1),

• wherein the engineered human HSCs or HSPCs are produced by transfecting human hemogenic endothelial cells (HE) in vitro with a non-integrative vector encoding the transcription factors ERG, HOXA9, HOXA5, LCOR and RUNX1,
• wherein the transcription factors are expressed in the HE cells to produce a population of multilineage HSCs and HSPCs capable of engrafting in the subject after implantation or transplantation,
• wherein the engineered HSCs or HSPCs are autologous or allogeneic to the subject;
• and
• wherein the subject is a patient who has undergone chemotherapy or irradiation, or both, and comprises deficiencies in immune function or lymphocyte reconstitution, or both deficiencies in immune function and lymphocyte reconstitution.