Reprogramming of mouse somatic cells into pluripotent stem-like cells using a combination of small molecules

Kang, Phil Jun; Moon, Jung Hwan; Yoon, Byung Sun; Hyeon, Solji; Jun, Eun Kyoung; Park, Gyuman; Yun, Wonjin; Park, Jiyong; Park, Minji; Kim, Aeree; Whang, Kwang Youn; Koh, Gou Young; Oh, Sejong; You, Seungkwon

doi:10.1016/j.biomaterials.2014.05.015

Cited by 33 publications

(24 citation statements)

References 39 publications

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“…Despite the improved expansion of iKCs and successful construction of a human skin equivalent in vitro, clinicians have continually questioned their value versus the risks of retrovirus-mediated gene transfer, which can potential contribute to undesired genotoxicity, insertional mutagenesis, and tumorigenesis [59]. Various integration-free approaches have been developed to convert cells into iPSCs and adult stem/progenitor cells, including viral (adenoviruses, Sendai viruses, and Creexcisable viruses) [60][61][62][63] and non-viral (DNA expression vectors, minicircle vectors, and episomal vectors) [64][65][66][67] vectors and non-DNA-based systems (proteins, mRNAs, and chemicals) [68][69][70][71][72][73]. Thus, further studies of genetic delivery that avoids potential issues associated with viral integration are required to increase the feasibility of clinical trials.…”

Section: Discussionmentioning

confidence: 99%

Long-term expansion of directly reprogrammed keratinocyte-like cells and in vitro reconstitution of human skin

et al. 2020

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Background: Human keratinocytes and derived products are crucial for skin repair and regeneration. Despite substantial advances in engineered skin equivalents, their poor availability and immunorejection remain major challenges in skin grafting. Methods: Induced keratinocyte-like cells (iKCs) were directly reprogrammed from human urine cells by retroviral transduction of two lineage-specific transcription factors BMI1 and △NP63α (BN). Expression of keratinocyte stem cell or their differentiation markers were assessed by PCR, immunofluorescence and RNA-Sequencing. Regeneration capacity of iKCs were assessed by reconstitution of a human skin equivalent under air-interface condition. Results: BN-driven iKCs were similar to primary keratinocytes (pKCs) in terms of their morphology, protein expression, differentiation potential, and global gene expression. Moreover, BN-iKCs self-assembled to form stratified skin equivalents in vitro. Conclusions: This study demonstrated an approach to generate human iKCs that could be directly reprogrammed from human somatic cells and extensively expanded in serum-and feeder cell-free systems, which will facilitate their broad applicability in an efficient and patient-specific manner.

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Section: Discussionmentioning

confidence: 99%

Long-term expansion of directly reprogrammed keratinocyte-like cells and in vitro reconstitution of human skin

et al. 2020

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“…Although the use of direct protein mediated method could completely avoid this problem, the reprogramming efficiency was very low (Kim et al, ), so there was not much practical value. Currently, induced pluripotent stem cells (CiPSC) could be successfully reprogrammed by small‐molecule compounds from mouse somatic cells which had higher reprogramming efficiency and no exogenous genes integration (Hou et al, ; Kang et al, ; Yang et al, ; Ye et al, ). It could be the most ideal method for iPSCs generation.…”

Section: Introductionmentioning

confidence: 99%

The modification of mitochondrial energy metabolism and histone of goat somatic cells under small molecules compounds induction

Zhang

Ren

et al. 2019

Reprod Domestic Animals

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In recent years, induced pluripotent stem cells (iPSCs) technique is able to allow us to generate pluripotency from somatic cells in vitro through the over expression of several transcription factors. Normally, viral vectors and transcription factors are commonly used on iPSC technique, which could cause many barriers on further application. In this study, we attempt to process a new method to obtain pluripotency from goat somatic cells in vitro under fully chemically defined condition. The results showed that chemically induced pluripotent stem cells-like cells (CiPSC-like cells) colonies were generated from goat ear fibroblasts by fully small-molecule compounds. Those three dimensions colonies were similar with mouse iPSCs in morphology and had strong positive alkaline phosphatase (AP) activity and expressed pluripotency related genes OCT4, SOX2, NANOG, CDH1, TDGF, GDF3, DAX1, REX1, which determined by RT-PCR. Those colonies could also differentiate into different cell types derived from three germ layers proved by RT-PCR and immunofluorescence assays. The expression of glycolysis related genes about PGAM1, KPYM2 and HXK2 in CiPSC-like colonies formation groups were significantly higher than their parental fibroblasts, but not in the non-CiPSC-like colonies formation group. The expression of histone acetylation and methylation related genes, HAT1 and SMYD3, was not significantly up-regulated within different groups compared to their parental fibroblasts, respectively. However, the expression of histone methylation related gene, KDM5B, was significantly up-regulated on the cells from non-colonies formation group compared to parental fibroblasts, but the expression of KDM5B of the cells from CiPSC-like cell colonies was not significantly difference compared to that of parental fibroblasts. In conclusion, this is the first report that CiPSC-like cells could be generated in vitro from goat rather than just mouse under fully chemically defined condition. The generation of CiPSC-like colonies may be depended on the correct modification of energy metabolism and histone epigenetic during the reprogramming, rather than just the over-expression of those pluripotency related genes. This study will strongly support us to further establish the stable goat CiPSC lines without any integration of exogenous genes. This article is protected by copyright. All rights reserved.

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“…Since the tissue-dependent expression of such transporter molecules is highly heterogeneous (see e.g., almost any dataset in the human protein atlas http://proteinatlas.org/ Uhlén et al, 2015, including those for SLC28 http://www.proteinatlas.org/search/slc28 and SLC29 http://www.proteinatlas.org/search/slc29), it must be subject to regulation (e.g., Pennycooke et al, 2001; Del Santo et al, 2001; Fernández-Veledo et al, 2004, 2007; Plant, 2016). Thus, just as with the small-molecule-driven induction of pluripotent stem cells (Okita et al, 2007; Feng et al, 2009; Desponts and Ding, 2010; Li and Ding, 2010; Zhang, 2010; Grskovic et al, 2011; Li et al, 2012, 2014; Li X. et al, 2015; Jung et al, 2014; Kang et al, 2014), that regulation can similarly be affected by pharmacological intervention with small molecule effectors. Thus, our aim was to seek small molecules that were themselves without cytotoxic effects but that could increase the response of different target cells to anti-cancer drugs that are otherwise present at only a barely cytotoxic level, in particular by modulating the level of activities of specific uptake transporters.…”

Section: Introductionmentioning

confidence: 99%

Enhancing Drug Efficacy and Therapeutic Index through Cheminformatics-Based Selection of Small Molecule Binary Weapons That Improve Transporter-Mediated Targeting: A Cytotoxicity System Based on Gemcitabine

et al. 2017

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The transport of drug molecules is mainly determined by the distribution of influx and efflux transporters for which they are substrates. To enable tissue targeting, we sought to develop the idea that we might affect the transporter-mediated disposition of small-molecule drugs via the addition of a second small molecule that of itself had no inhibitory pharmacological effect but that influenced the expression of transporters for the primary drug. We refer to this as a “binary weapon” strategy. The experimental system tested the ability of a molecule that on its own had no cytotoxic effect to increase the toxicity of the nucleoside analog gemcitabine to Panc1 pancreatic cancer cells. An initial phenotypic screen of a 500-member polar drug (fragment) library yielded three “hits.” The structures of 20 of the other 2,000 members of this library suite had a Tanimoto similarity greater than 0.7 to those of the initial hits, and each was itself a hit (the cheminformatics thus providing for a massive enrichment). We chose the top six representatives for further study. They fell into three clusters whose members bore reasonable structural similarities to each other (two were in fact isomers), lending strength to the self-consistency of both our conceptual and experimental strategies. Existing literature had suggested that indole-3-carbinol might play a similar role to that of our fragments, but in our hands it was without effect; nor was it structurally similar to any of our hits. As there was no evidence that the fragments could affect toxicity directly, we looked for effects on transporter transcript levels. In our hands, only the ENT1-3 uptake and ABCC2,3,4,5, and 10 efflux transporters displayed measurable transcripts in Panc1 cultures, along with a ribonucleoside reductase RRM1 known to affect gemcitabine toxicity. Very strikingly, the addition of gemcitabine alone increased the expression of the transcript for ABCC2 (MRP2) by more than 12-fold, and that of RRM1 by more than fourfold, and each of the fragment “hits” served to reverse this. However, an inhibitor of ABCC2 was without significant effect, implying that RRM1 was possibly the more significant player. These effects were somewhat selective for Panc cells. It seems, therefore, that while the effects we measured were here mediated more by efflux than influx transporters, and potentially by other means, the binary weapon idea is hereby fully confirmed: it is indeed possible to find molecules that manipulate the expression of transporters that are involved in the bioactivity of a pharmaceutical drug. This opens up an entirely new area, that of chemical genomics-based drug targeting.

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Reprogramming of mouse somatic cells into pluripotent stem-like cells using a combination of small molecules

Cited by 33 publications

References 39 publications

Long-term expansion of directly reprogrammed keratinocyte-like cells and in vitro reconstitution of human skin

Long-term expansion of directly reprogrammed keratinocyte-like cells and in vitro reconstitution of human skin

The modification of mitochondrial energy metabolism and histone of goat somatic cells under small molecules compounds induction

Enhancing Drug Efficacy and Therapeutic Index through Cheminformatics-Based Selection of Small Molecule Binary Weapons That Improve Transporter-Mediated Targeting: A Cytotoxicity System Based on Gemcitabine

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