Cel Generating Hematopoietic Stem Cells (HSCs) by Transcription Factor (TF) overexpression is of great interest in the stem cell field and promises considerable therapeutic potential. Among the systems described so far only one study, using murine B cells, has led to the generation of transplantable induced HSCs (iHSCs), although at exceedingly low efficiencies and using combinations between 6 and 8 TFs. We propose here a new approach to generate iHSCs based on the use of newly identified highly plastic “elite” type cells for reprogramming by the Yamanaka factors into induced pluripotent stem cells, generated by the transient exposure of B cells to C/EBPα. Hypothesizing that microRNAs could potentiate the effect of TFs in HSC reprogramming we will also analyze the effects of microRNA overexpression. For this we will first analyze HSCs and different hematopoietic progenitors to identify differentially expressed microRNAs. Different combinations of selected microRNAs will be cloned into inducible retroviral vectors together with hematopoietic TFs and used to infect “elite” CEBPα-primed B cells to screen for the induction, in vitro and in vivo, of iHSCs. The use of colony assays, cell surface marker and transcriptomic analyses of the resulting cells, as well as transplantation into irradiated mice, will enable us to identify microRNAs and TFs combinations that induce HSC reprogramming. The heterogeneity of iHSCs will then be tested by single cell expression analysis using RNA-seq. This project will provide new insights into the reprogramming of HSCs, the clinically most important type of adult stem cells, with potential medical applications. The proposed action will benefit the applicant’s career through acquisition of scientific maturity and independence. Dziedzina nauki medical and health sciencesmedical biotechnologycells technologiesstem cellsmedical and health sciencesclinical medicineoncologymedical and health sciencesclinical medicinetransplantationnatural sciencesbiological sciencesgeneticschromosomesnatural sciencesbiological sciencesgeneticsgenomes Program(-y) H2020-EU.1.3. - EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions Main Programme H2020-EU.1.3.2. - Nurturing excellence by means of cross-border and cross-sector mobility Temat(-y) MSCA-IF-2016 - Individual Fellowships Zaproszenie do składania wniosków H2020-MSCA-IF-2016 Zobacz inne projekty w ramach tego zaproszenia System finansowania MSCA-IF-EF-ST - Standard EF Koordynator FUNDACIO CENTRE DE REGULACIO GENOMICA Wkład UE netto € 158 121,60 Adres CARRER DOCTOR AIGUADER 88 08003 Barcelona Hiszpania Zobacz na mapie Region Este Cataluña Barcelona Rodzaj działalności Research Organisations Linki Kontakt z organizacją Opens in new window Strona internetowa Opens in new window Uczestnictwo w unijnych programach w zakresie badań i innowacji Opens in new window sieć współpracy HORIZON Opens in new window Koszt całkowity € 158 121,60
