title: SOX2 and SOX2-MYC Reprogramming Process of Fibroblasts to the Neural Stem Cells Compromised by Senescence.
abstract: Tumorigenic potential of induced pluripotent stem cells (iPSCs) infiltrating population of induced neural stem cells (iNSCs) generated from iPSCs may limit their medical applications. To overcome such a difficulty, direct reprogramming of adult somatic cells into iNSCs was proposed. The aim of this study was the systematic comparison of induced neural cells (iNc) obtained with different methods-direct reprogramming of human adult fibroblasts with either SOX2 (SiNSc-like) or SOX2 and c-MYC (SMiNSc-like) and induced pluripotent stem cells differentiation to ebiNSc-in terms of gene expression profile, differentiation potential as well as proliferation properties. Immunocytochemistry and real-time PCR analyses were used to evaluate gene expression profile and differentiation potential of various iNc types. Bromodeoxyuridine (BrdU) incorporation and senescence-associated beta-galactosidase (SA-β-gal) assays were used to estimate proliferation potential. All three types of iNc were capable of neuronal differentiation; however, astrocytic differentiation was possible only in case of ebiNSc. Contrary to ebiNSc generation, the direct reprogramming was rarely a propitious process, despite 100% transduction efficiency. The potency of direct iNSCs-like cells generation was lower as compared to iNSCs obtained by iPSCs differentiation, and only slightly improved when c-MYC was added. Directly reprogrammed iNSCs-like cells were lacking the ability to differentiate into astrocytic cells and characterized by poor efficiency of neuronal cells formation. Such features indicated that these cells could not be fully reprogrammed, as confirmed mainly with senescence detection. Importantly, SiNSc-like and SMiNSc-like cells were unable to achieve the long-term survival and became senescent, which limits their possible therapeutic applicability. Our results suggest that iNSCs-like cells, generated in the direct reprogramming attempts, were either not fully reprogrammed or reprogrammed only into neuronal progenitors, mainly because of the inaccuracies of currently available protocols.
authors.affiliation: Department of Research and Development, Celther Polska, Lodz, Poland.|Department of Tumor Biology, Medical University of Lodz, Lodz, Poland.
authors.forename: Marta
authors.initials: M
authors.lastname: Winiecka-Klimek
authors.affiliation: Department of Research and Development, Celther Polska, Lodz, Poland.|Department of Tumor Biology, Medical University of Lodz, Lodz, Poland.
authors.forename: Maciej
authors.initials: M
authors.lastname: Smolarz
authors.affiliation: Department of Research and Development, Celther Polska, Lodz, Poland.
authors.forename: Maciej P
authors.initials: MP
authors.lastname: Walczak
authors.affiliation: Department of Research and Development, Celther Polska, Lodz, Poland.|Department of Tumor Biology, Medical University of Lodz, Lodz, Poland.
authors.forename: Jolanta
authors.initials: J
authors.lastname: Zieba
authors.affiliation: Department of Research and Development, Celther Polska, Lodz, Poland.
authors.forename: Krystyna
authors.initials: K
authors.lastname: Hulas-Bigoszewska
authors.affiliation: Department of Medical Law, Chair of Human Sciences, Medical University of Lodz, Lodz, Poland.
authors.forename: Blazej
authors.initials: B
authors.lastname: Kmieciak
authors.affiliation: Department of Research and Development, Celther Polska, Lodz, Poland.|Department of Tumor Biology, Medical University of Lodz, Lodz, Poland.
authors.forename: Sylwester
authors.initials: S
authors.lastname: Piaskowski
authors.affiliation: Department of Research and Development, Celther Polska, Lodz, Poland.|Department of Tumor Biology, Medical University of Lodz, Lodz, Poland.
authors.forename: Piotr
authors.initials: P
authors.lastname: Rieske
authors.affiliation: Department of Research and Development, Celther Polska, Lodz, Poland.
authors.forename: Dawid P
authors.initials: DP
authors.lastname: Grzela
authors.affiliation: Department of Research and Development, Celther Polska, Lodz, Poland.|Department of Tumor Biology, Medical University of Lodz, Lodz, Poland.
authors.forename: Ewelina
authors.initials: E
authors.lastname: Stoczynska-Fidelus
mesh_terms: D065150:Cellular Reprogramming; D000067470:Cellular Reprogramming Techniques / Q000379:methods; D016922:Cellular Senescence; D005347:Fibroblasts / Q000378:metabolism*; D006801:Humans; D058953:Neural Stem Cells / Q000378:metabolism*; D016271:Proto-Oncogene Proteins c-myc / Q000096:biosynthesis* / Q000235:genetics; D055748:SOXB1 Transcription Factors / Q000096:biosynthesis* / Q000235:genetics; D014161:Transduction, Genetic / Q000379:methods
chemical_list: C489427:MYC protein, human; D016271:Proto-Oncogene Proteins c-myc; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors