Single Transcription Factor Conversion of Human Blood Fate to NPCs with CNS and PNS Developmental Capacity

Lee, Jonghee; Mitchell, Ryan R.; McNicol, Jamie D.; Shapovalova, Zoya; Laronde, Sarah; Tanasijevic, Borko; Milsom, Chlöe; Casado, Fanny L.; Fiebig‐Comyn, Aline; Collins, Tony; Singh, Karun K.; Bhatia, Mickie

doi:10.1016/j.celrep.2015.04.056

Cited by 69 publications

(87 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…In 2012, Kumar et al [27] generated human fibroblastderived iNSCs through retroviral overexpression of Brn2 with Klf4, Sox2, and Zic3 in a medium supplemented with growth factor FGF2. Two studies reported that lentivirus-mediated expression of OCT4 is sufficient to obtain iNSCs from adult human fibroblasts, neonatal cord, and peripheral blood cells [29,30]. Two studies reported that lentivirus-mediated expression of OCT4 is sufficient to obtain iNSCs from adult human fibroblasts, neonatal cord, and peripheral blood cells [29,30].…”

Section: Transcription Factors Employed In Insc Generationmentioning

confidence: 99%

Take the shortcut – direct conversion of somatic cells into induced neural stem cells and their biomedical applications

et al. 2019

View full text Add to dashboard Cite

Second-generation reprogramming of somatic cells directly into the cell type of interest avoids induction of pluripotency and subsequent cumbersome differentiation procedures. Several recent studies have reported direct conversion of human somatic cells into stably proliferating induced neural stem cells (iNSCs). Importantly, iNSCs are easier, faster, and more cost-efficient to generate than induced pluripotent stem cells (iPSCs), and also have a higher level of clinical safety. Stably, self-renewing iNSCs can be derived from different cellular sources, such as skin fibroblasts and peripheral blood mononuclear cells, and readily differentiate into neuronal and glial lineages that are indistinguishable from their iPSC-derived counterparts or from NSCs isolated from primary tissues. This review focuses on the derivation and characterization of iNSCs and their biomedical applications. We first outline different approaches to generate iNSCs and then discuss the underlying molecular mechanisms. Finally, we summarize the preclinical validation of iNSCs to highlight that these cells are promising targets for disease modeling, autologous cell therapy, and precision medicine. Take the shortcut: from iPSC to iNSCForced expression of lineage-specific transcription factors (TFs) has been shown to reprogram the developmental potential of various somatic cell types e.g. by inducing pluripotency [1]. The seminal breakthrough of fibroblast reprogramming into iPSCs offers a novel experimental tool to generate patient-specific cells for developmental studies and diverse biomedical applications, such as disease modeling and cell therapy. Since then, efficiency and robustness of reprogramming protocols have been substantially improved, but the derivation of patient-specific iPSCs remains a lengthy and cumbersome procedure. Moreover, clinical applications require subsequent redifferentiation of iPSCs into the cell type of interest, which is inefficient and risky because transplanted undifferentiated iPSCs are tumorigenic. In the shadow of iPSC-type reprogramming, second-generation reprogramming paradigms have been developed to bypass the pluripotency state Abbreviations

show abstract

Section: Transcription Factors Employed In Insc Generationmentioning

confidence: 99%

Take the shortcut – direct conversion of somatic cells into induced neural stem cells and their biomedical applications

et al. 2019

View full text Add to dashboard Cite

show abstract

“…Recently, induced neural progenitor cells (iNPCs) from human neonatal and adult peripheral blood were generated using a single transcription factor OCT4 combined with SMAD+GSK-3 inhibition. Blood-derived iNPCs differentiated in vivo and responded to guided differentiation in vitro into glia (astrocytes and oligodendrocytes) and multiple neuronal subtypes, including dopaminergic and nociceptive neurons (Lee et al, 2015). Furthermore, nociceptive neurons obtained from differentiated iNPCs phenocopy chemotherapy-induced neurotoxicity in a system suitable for high-throughput drug screening and therefore harbor great utility for the study of clinically relevant neurological diseases directly from patient samples (Lee et al, 2015).…”

Section: Direct Reprogramming As a Tool To Derive Functional Neurons mentioning

confidence: 99%

“…Blood-derived iNPCs differentiated in vivo and responded to guided differentiation in vitro into glia (astrocytes and oligodendrocytes) and multiple neuronal subtypes, including dopaminergic and nociceptive neurons (Lee et al, 2015). Furthermore, nociceptive neurons obtained from differentiated iNPCs phenocopy chemotherapy-induced neurotoxicity in a system suitable for high-throughput drug screening and therefore harbor great utility for the study of clinically relevant neurological diseases directly from patient samples (Lee et al, 2015). Tian et al recently demonstrated the direct conversion of mouse fibroblasts into lineage restricted induced dopaminergic precursors by ectopic expression of Brn2, Sox2 and Foxa2 (Tian et al, 2015).…”

Section: Direct Reprogramming As a Tool To Derive Functional Neurons mentioning

confidence: 99%

New approaches for direct conversion of patient fibroblasts into neural cells

2017

View full text Add to dashboard Cite

Recent landmark studies have demonstrated the production of disease-relevant human cell types by two different methods; differentiation of stem cells using external morphogens or lineage conversion using genetic factors. Directed differentiation changes embryonic stem cells (ESCs) or induced pluripotent stem cells (iPSCs) into a desired cell type by providing developmental cues in an in vitro environment. Direct reprogramming is achieved by the introduction of exogenous lineage specific transcription factors to convert any somatic cell type into another, thereby bypassing an intermediate pluripotent stage. A variety of somatic cell types such as blood, keratinocytes and fibroblasts can be used to derive iPSC cells. However, the process is time consuming, laborious, expensive and gives rise to cells with reported epigenetic heterogeneity even amongst different iPSC lines from same patient which could propagate phenotypic variability. A major concern with the use of pluripotent cells as starting material for cell replacement therapy is their incomplete differentiation and their propensity to form tumors following transplantation. In comparison, transcription factor mediated reprogramming offers a direct route to target cell types. This could allow for rapid comparison of large cohorts of patient and control samples at a given time for disease modeling. Additionally, transcription factors that drive maturation may yield more functionally mature cells than directed differentiation. Several studies have demonstrated the feasibility of generating of cell types such as cardiomyocytes, hepatocytes, and neurons from fibroblasts. Here, we will discuss recent advances and key challenges regarding direct reprogramming of somatic cell types into diverse neural cells.

show abstract

“…Taken together, these findings clearly suggest that e-iNSCs might be a safer cell source than r-iNSCs as well as NSCs derived from pluripotent stem cells. A recent study demonstrated the derivation of iNSCs from blood cells (44). As blood cells are one of the easily accessible autologous cell sources, further studies will aim to assess whether our episomal vector system could affect the direct conversion of human blood cells, and other human somatic cells, into iNSCs.…”

Section: Discussionmentioning

confidence: 99%

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

Kim

Kwak³

et al. 2016

Journal of Biological Chemistry

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

Single Transcription Factor Conversion of Human Blood Fate to NPCs with CNS and PNS Developmental Capacity

Cited by 69 publications

References 35 publications

Take the shortcut – direct conversion of somatic cells into induced neural stem cells and their biomedical applications

Take the shortcut – direct conversion of somatic cells into induced neural stem cells and their biomedical applications

New approaches for direct conversion of patient fibroblasts into neural cells

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

Contact Info

Product

Resources

About