title: Immunogenic Dendritic Cell Generation from Pluripotent Stem Cells by Ectopic Expression of Runx3.
abstract: Application of dendritic cells (DCs) to prime responses to tumor Ags provides a promising approach to immunotherapy. However, only a limited number of DCs can be manufactured from adult precursors. In contrast, pluripotent embryonic stem (ES) cells represent an inexhaustible source for DC production, although it remains a major challenge to steer directional differentiation because ES cell-derived cells are typically immature with impaired functional capacity. Consistent with this notion, we found that mouse ES cell-derived DCs (ES-DCs) represented less mature cells compared with bone marrow-derived DCs. This finding prompted us to compare the gene expression profile of the ES cell- and adult progenitor-derived, GM-CSF-instructed, nonconventional DC subsets. We quantified the mRNA level of 17 DC-specific transcription factors and observed that 3 transcriptional regulators (Irf4, Spi-B, and Runx3) showed lower expression in ES-DCs than in bone marrow-derived DCs. In light of this altered gene expression, we probed the effects of these transcription factors in developing mouse ES-DCs with an isogenic expression screen. Our analysis revealed that forced expression of Irf4 repressed ES-DC development, whereas, in contrast, Runx3 improved the ES-DC maturation capacity. Moreover, LPS-treated and Runx3-activated ES-DCs exhibited enhanced T cell activation and migratory potential. In summary, we found that ex vivo-generated ES-DCs had a compromised maturation ability and immunogenicity. However, ectopic expression of Runx3 enhances cytokine-driven ES-DC development and acts as an instructive tool for the generation of mature DCs with enhanced immunogenicity from pluripotent stem cells.
authors.affiliation: Stem Cell Differentiation Laboratory, Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, H-4010 Debrecen, Hungary.
authors.forename: Erika
authors.initials: E
authors.lastname: Takacs
authors.affiliation: Stem Cell Differentiation Laboratory, Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, H-4010 Debrecen, Hungary.
authors.forename: Pal
authors.identifier: 0000-0002-8780-5199
authors.initials: P
authors.lastname: Boto
authors.affiliation: Stem Cell Differentiation Laboratory, Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, H-4010 Debrecen, Hungary.
authors.forename: Emilia
authors.initials: E
authors.lastname: Simo
authors.affiliation: Stem Cell Differentiation Laboratory, Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, H-4010 Debrecen, Hungary.
authors.forename: Tamas I
authors.initials: TI
authors.lastname: Csuth
authors.affiliation: Stem Cell Differentiation Laboratory, Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, H-4010 Debrecen, Hungary.
authors.forename: Bianka M
authors.initials: BM
authors.lastname: Toth
authors.affiliation: Biotalentum Ltd., H-2100 Gödöllő, Hungary.
authors.forename: Hadas
authors.identifier: 0000-0003-3300-8597
authors.initials: H
authors.lastname: Raveh-Amit
authors.affiliation: Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, H-4012 Debrecen, Hungary.
authors.forename: Attila
authors.initials: A
authors.lastname: Pap
authors.affiliation: Department of Physiology, Faculty of Medicine, University of Debrecen, H-4010 Debrecen, Hungary.
authors.forename: Elek G
authors.initials: EG
authors.lastname: Kovács
authors.affiliation: Biotalentum Ltd., H-2100 Gödöllő, Hungary.
authors.forename: Julianna
authors.initials: J
authors.lastname: Kobolak
authors.affiliation: Department of Physiology, Faculty of Medicine, University of Debrecen, H-4010 Debrecen, Hungary.
authors.forename: Szilvia
authors.identifier: 0000-0001-7356-6345
authors.initials: S
authors.lastname: Benkö
authors.affiliation: Biotalentum Ltd., H-2100 Gödöllő, Hungary.|Molecular Animal Biotechnology Laboratory, Szent István University, H-2101 Gödöllő, Hungary; and.
authors.forename: Andras
authors.identifier: 0000-0003-3791-2583
authors.initials: A
authors.lastname: Dinnyes
authors.affiliation: Stem Cell Differentiation Laboratory, Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, H-4010 Debrecen, Hungary; szatmari@med.unideb.hu.|Faculty of Pharmacy, University of Debrecen, H-4032 Debrecen, Hungary.
authors.forename: Istvan
authors.initials: I
authors.lastname: Szatmari
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002454:Cell Differentiation / Q000502:physiology*; D002469:Cell Separation; D002478:Cells, Cultured; D050677:Core Binding Factor Alpha 3 Subunit / Q000096:biosynthesis* / Q000276:immunology; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D000066630:Ectopic Gene Expression / Q000502:physiology*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D008213:Lymphocyte Activation / Q000276:immunology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D059467:Transcriptome
chemical_list: D050677:Core Binding Factor Alpha 3 Subunit; C493730:Runx3 protein, mouse