Direct reprogramming of mouse fibroblasts to neural progenitors. Proc Natl Acad Sci USA

Department of Chemistry, The Scripps Research Institute, La Jolla, CA 92037, USA.
Proceedings of the National Academy of Sciences (Impact Factor: 9.67). 05/2011; 108(19):7838-43. DOI: 10.1073/pnas.1103113108
Source: PubMed


The simple yet powerful technique of induced pluripotency may eventually supply a wide range of differentiated cells for cell therapy and drug development. However, making the appropriate cells via induced pluripotent stem cells (iPSCs) requires reprogramming of somatic cells and subsequent redifferentiation. Given how arduous and lengthy this process can be, we sought to determine whether it might be possible to convert somatic cells into lineage-specific stem/progenitor cells of another germ layer in one step, bypassing the intermediate pluripotent stage. Here we show that transient induction of the four reprogramming factors (Oct4, Sox2, Klf4, and c-Myc) can efficiently transdifferentiate fibroblasts into functional neural stem/progenitor cells (NPCs) with appropriate signaling inputs. Compared with induced neurons (or iN cells, which are directly converted from fibroblasts), transdifferentiated NPCs have the distinct advantage of being expandable in vitro and retaining the ability to give rise to multiple neuronal subtypes and glial cells. Our results provide a unique paradigm for iPSC-factor-based reprogramming by demonstrating that it can be readily modified to serve as a general platform for transdifferentiation.

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Available from: Janghwan Kim,
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    • "More recently, somatic cells were redirected into another differentiated cell lineage, without passing an intermediate pluripotent stage, by transducing a set of transcription factors that play crucial regulatory roles in the differentiation of the destination cell; direct conversion , or direct reprogramming, of murine fibroblasts into cardiomyocytes (Ieda et al., 2010; Inagawa et al., 2012; Inagawa and Ieda, 2013), neurons (Kim et al., 2011, 2012; Caiazzo et al., 2011; Han et al., 2012), chondrocytes (Hiramatsu et al., 2011), and hepatocytes (Sekiya and Suzuki, 2011; Huang et al., 2011), as well as of human fibroblasts into cardiomyocytes (Wada et al., 2013; Nam et al., 2013), neurons (Pang et al., 2011; Caiazzo et al., 2011; Kim et al., 2012), and hematopoietic cells (Szabo et al., 2010) have been reported. Although the efficiencies of direct conversion were generally low (0.005%–30% of fibroblasts were successfully converted into the cells of interest; Thier et al., 2012; Sekiya and Suzuki, 2011; Pang et al., 2011; Nam et al., 2013; Kim et al., 2011, 2012; Inagawa et al., 2012; Inagawa and Ieda, 2013; Ieda et al., 2010; Huang et al., 2011; Han et al., 2012; Caiazzo et al., 2011), these technologies may be quite valuable for generation of the desired cells that could be used in basic research as well as in regenerative therapy for various human disorders. We have recently succeeded in directly reprogramming human fibroblasts into osteoblasts that massively produced bone matrix and contributed to bone regeneration with an efficiency as high as 80% (Yamamoto et al., 2015). "
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    ABSTRACT: Brown adipocytes (BAs) play important roles in body temperature regulation, energy balance, and carbohydrate and lipid metabolism. Activities of BAs are remarkably diminished in obese and diabetic patients, providing possibilities of transplanting functional BAs resulting in therapeutic benefit. Here, we show generation of functional BAs by cellular reprogramming procedures. Transduction of the PRDM16 gene into iPSC-derived embryoid bodies induced BA phenotypes (iBAs). Moreover, normal human fibroblasts were directly converted into BAs (dBAs) by C/EBP-β and C-MYC gene transduction. Approximately 90% of the fibroblasts were successfully converted within 12 days. The dBAs were highly active in mitochondrial biogenesis and oxidative metabolism. Mouse dBAs were induced by Prdm16, C/ebp-β, and L-myc genes, and after transplantation, they significantly reduced diet-induced obesity and insulin resistance in an UCP1-dependent manner. Thus, highly functional BAs can be generated by cellular reprogramming, suggesting a promising tailor-made cell therapy against metabolic disorders including type 2 diabetes mellitus.
    Stem Cell Reports 09/2015; 5(4). DOI:10.1016/j.stemcr.2015.08.007 · 5.37 Impact Factor
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    • "plasticity induction can be achieved without additional TFs (Efe et al., 2011; J. Kim et al., 2011; J. Li et al., 2013) or small molecules (H. Wang et al., 2014; Zhu et al., 2014). "
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    ABSTRACT: The combination of OCT4 expression and short-term exposure to reprogramming media induces a state of transcriptional plasticity in human fibroblasts, capable of responding to changes in the extracellular environment that facilitate direct cell fate conversion toward lineage specific progenitors. Here we reveal that continued exposure of OCT4-induced plastic human fibroblasts to reprogramming media (RM) is sufficient to induce pluripotency. OCT4-derived induced pluripotent stem cell (iPSC(OCT4)) colonies emerged after prolonged culture in RM, and formed independently of lineage specific progenitors. Human iPSC(OCT4) are morphologically indistinguishable from conventionally derived iPSCs and express core proteins involved in maintenance of pluripotency. iPSC(OCT4) display in vivo functional pluripotency as measured by teratoma formation consisting of the three germ layers, and are capable of targeted in vitro differentiation. Our study indicates that acquisition of pluripotency is one of multiple cell fate choices that can be facilitated through environmental stimulation of OCT4-induced plasticity, and suggests the role of other reprogramming factors to induce pluripotency can be substituted by prolonged culture of plastic fibroblasts. Copyright © 2015. Published by Elsevier B.V.
    Stem Cell Research 06/2015; 8(1). DOI:10.1016/j.scr.2015.06.006 · 3.69 Impact Factor
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    • "Because such therapies need large numbers of iNCs, induced mature neurons are terminally differentiated neurons that are unable to expand. Alternatively, neural progenitors or immature neural cells prepared in vitro might generate different neural lineages in vivo including mature neurons (Kim et al., 2011). Our results indicate that p53 deficiency induces three neural lineages and coculture of iNCs with primary neurons could enhance neuron maturation, suggesting that targeting the p53 pathway to generate functional neural tissues in vivo might be especially important. "
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    ABSTRACT: Differentiated somatic cells have been reprogrammed to a pluripotent state by forced expression of a set of transcription factors (Takahashi et al., 2007), indicating that terminally differentiated cells can be induced to undergo cell fate change. Recent studies further demonstrated that direct conversion from fibroblast to neuron, a potential cell replacement therapy for neurological disorders, can be induced by a set of transcription factors without passing through a pluripotent state (Caiazzo et al., 2011, Vierbuchen et al., 2010, Pfisterer et al., 2011, Pang et al., 2011, Yoo et al., 2011 and Ambasudhan et al., 2011). However, the mechanism underlying this conversion process remains largely unclear. As a result, a variety of combinations of transcription factors have been tried but generally with low percentages and very slow time course of conversion.
    Cell Reports 12/2014; 9(6). DOI:10.1016/j.celrep.2014.11.040 · 8.36 Impact Factor
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