Therapeutic Potential of Induced Neural Stem Cells for Spinal Cord Injury

Hong, Jongbae; Lee, Sung Ho; Lee, Seung Chan; Kim, Jongwan; Kim, Kee-Pyo; Kim, Sung Min; Tàpia, Natalia; Lim, Kyung Tae; Kim, Jong Hun; Ahn, Hong Sun; Ko, Kinarm; Shin, Chan Young; Lee, Hoon Taek; Schöler, Hans R.; Hyun, Jung Keun; Han, Dong Wook

doi:10.1074/jbc.m114.588871

Cited by 65 publications

(77 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…6C). As in our previous transplantation studies (13,20), here we found no obvious evidence for tumor formation in any of the transplanted rats (data not shown). To further investigate the fate of the engrafted e-iNSCs, we checked for the presence of Nestin, a progenitor marker, but found no GFP-positive cells expressing Nestin, indicating that the engrafted e-iNSCs had lost their stem cell identity (Fig.…”

Section: Single Transfection Of Episomal Vectors Is Sufficient For Gesupporting

confidence: 89%

“…Finally, to understand this gradual process on the epigenetic level, we next checked the DNA methylation status of both Nestin, a NSC marker (19), and Col1a1, a fibroblast marker (13). Notably, the promoter region of Col1a1 was dramatically de novo methylated even in the early passage e-iNSCs to the level similar to control NSCs (Fig.…”

Section: Single Transfection Of Episomal Vectors Is Sufficient For Gementioning

confidence: 99%

“…They could also differentiate into neurons, astrocytes, and oligodendrocytes both in vitro and in vivo. Furthermore, in a recent preclinical study, iNSCs were shown to have therapeutic potential following transplantation into the spinal cord injury disease model (13).…”

mentioning

confidence: 99%

See 2 more Smart Citations

Generation of Integration-free Induced Neural Stem Cells from Mouse Fibroblasts

Kim

Kwak³

et al. 2016

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

The viral vector-mediated overexpression of the defined transcription factors, Brn4/Pou3f4, Sox2, Klf4, and c-Myc (BSKM), could induce the direct conversion of somatic fibroblasts into induced neural stem cells (iNSCs). However, viral vectors may be randomly integrated into the host genome thereby increasing the risk for undesired genotoxicity, mutagenesis, and tumor formation. Here we describe the generation of integration-free iNSCs from mouse fibroblasts by non-viral episomal vectors containing BSKM. The episomal vector-derived iNSCs (eiNSCs) closely resemble control NSCs, and iNSCs generated by retrovirus (r-iNSCs) in morphology, gene expression profile, epigenetic status, and self-renewal capacity. The e-iNSCs are functionally mature, as they could differentiate into all the neuronal cell types both in vitro and in vivo. Our study provides a novel concept for generating functional iNSCs using a non-viral, non-integrating, plasmid-based system that could facilitate their biomedical applicability. Neural stem cells (NSCs),3 somatic stem cells of the brain and spinal cord, could differentiate into three major cell types (neurons, astrocytes, and oligodendrocytes) in the central nervous system (CNS) in response to relevant signaling pathways (1). A number of in vitro transplantation studies with disease animal models have demonstrated that the engrafted NSCs can functionally rescue disease-related phenotypes, proving their therapeutic potential (2-5). Thus, NSCs have long been considered as a potential source for replacing damaged cells and tissues often found in various neurodegenerative diseases (2-5). However, major concerns associated with these cells may preclude expediting advances into their therapeutic application. First, the accessibility of NSCs is largely restricted in vitro, due to their limited origins. Second, the autologous cell transplantation is not feasible using NSCs from their in vivo origins, and furthermore, allogeneic transplantation of NSCs may raise the potential for immune rejection. Finally, with current culture conditions, it is technically challenging to homogeneously maintain human NSCs in vitro (6, 7). Thus, NSC-like cells generated from easily accessible patient somatic cell types such as fibroblasts and blood cells could serve as an autologous source for therapeutic applications, thereby overcoming current obstacles to NSC-mediated translation research.We and others have demonstrated the direct conversion of somatic fibroblasts into self-renewing and multipotent induced neural stem cells (iNSCs) or induced neural progenitor cells (iNPCs) by the forced expression of different sets of transcription factors (8 -12). Recently, Thier et al. (12) have shown that the restricted expression of Oct4 by a tetracycline-dependent lentiviral vector, together with retrovirus-mediated overexpression of Sox2, Klf4, and c-Myc, could elicit the conversion of fibroblasts into iNSCs. Simultaneously, we have generated multipotent and self-renewing iNSCs from mouse fibroblasts by retrovirus-mediated ...

show abstract

Section: Single Transfection Of Episomal Vectors Is Sufficient For Gesupporting

confidence: 89%

Section: Single Transfection Of Episomal Vectors Is Sufficient For Gementioning

confidence: 99%

See 1 more Smart Citation

Generation of Integration-free Induced Neural Stem Cells from Mouse Fibroblasts

Kim

Kwak³

et al. 2016

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…The bisulfite‐converted DNA was amplified by PCR with previously described primers 70, 71. The PCR products were cloned into the pCRII TOPO vector (Invitrogen), and plasmids extracted from individual clones were sequenced by GATC‐biotech (http://www.gatc-biotech.com/en/index.html).…”

Section: Methodsmentioning

confidence: 99%

Changing POU dimerization preferences converts Oct6 into a pluripotency inducer

Jerabek

et al. 2016

EMBO Reports

123

View full text Add to dashboard Cite

The transcription factor Oct4 is a core component of molecular cocktails inducing pluripotent stem cells (iPSCs), while other members of the POU family cannot replace Oct4 with comparable efficiency. Rather, group III POU factors such as Oct6 induce neural lineages. Here, we sought to identify molecular features determining the differential DNA‐binding and reprogramming activity of Oct4 and Oct6. In enhancers of pluripotency genes, Oct4 cooperates with Sox2 on heterodimeric SoxOct elements. By re‐analyzing ChIP‐Seq data and performing dimerization assays, we found that Oct6 homodimerizes on palindromic OctOct more cooperatively and more stably than Oct4. Using structural and biochemical analyses, we identified a single amino acid directing binding to the respective DNA elements. A change in this amino acid decreases the ability of Oct4 to generate iPSCs, while the reverse mutation in Oct6 does not augment its reprogramming activity. Yet, with two additional amino acid exchanges, Oct6 acquires the ability to generate iPSCs and maintain pluripotency. Together, we demonstrate that cell type‐specific POU factor function is determined by select residues that affect DNA‐dependent dimerization.

show abstract

“…Studies have recently addressed this gap within the literature, reporting the restorative effects of induced neural stem cells in models of spinal cord injury and Parkinson's disease. In one study, mouse embryonic fibroblast-derived induced neural stem cells were transplanted into the contused thoracic spinal cord of rats [151] . Following transplantation, induced neural stem cells lost their stem cell identity and differentiated into neurons, astrocytes, and oligodendrocytes, with synaptic formation observed between host and transplanted neurons.…”

Section: Clinical Applications Of Induced Neural Cell Typesmentioning

confidence: 99%