Direct Neural Conversion from Human Fibroblasts Using Self-Regulating and Nonintegrating Viral Vectors

Lau, Shong; Ottosson, Daniella Rylander; Jakobsson, Johan; Parmar, Malin

doi:10.1016/j.celrep.2014.11.017

Cited by 37 publications

(28 citation statements)

References 35 publications

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“…Many reports have demonstrated successfully that an inducible single factor mediated reprogramming generates highly efficient conversion (Chanda et al, 2014; Kim et al, 2014; Lujan et al, 2012). Other studies have also utilized the inducible transgene expression when using multiple factors (Cassady et al, 2014; Mazzoni et al, 2013), generating neuronal cells from human fibroblasts with inducible microRNAs (Lau et al, 2014) and induced dopaminergic neurons from mouse fibroblasts (Kim et al, 2011b). …”

Section: Challenges and New Directions In Deriving Direct Reprogrammementioning

confidence: 99%

New approaches for direct conversion of patient fibroblasts into neural cells

2017

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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.

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Section: Challenges and New Directions In Deriving Direct Reprogrammementioning

confidence: 99%

New approaches for direct conversion of patient fibroblasts into neural cells

2017

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“…To explore suitable approaches for neuronal generation, we and other groups have developed a non-integrating transgene system for achieving induced neuronal cells from fibroblasts (11,12). In such systems the desired cells can be directly obtained by the introduction of transcription factors, such as Ascl1, Brn2, and Ngn2.…”

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confidence: 99%

Conversion of Fibroblasts to Parvalbumin Neurons by One Transcription Factor, Ascl1, and the Chemical Compound Forskolin

Shi

Zhang

Chen

et al. 2016

Journal of Biological Chemistry

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Abnormalities in parvalbumin (PV)-expressing interneuronscause neurodevelopmental disorders such as epilepsy, autism, and schizophrenia. Unlike other types of neurons that can be efficiently differentiated from pluripotent stem cells, PV neurons were minimally generated using a conventional differentiation strategy. In this study we developed an adenovirus-based transdifferentiation strategy that incorporates an additional chemical compound for the efficient generation of induced PV (iPV) neurons. The chemical compound forskolin combined with Ascl1 induced ϳ80% of mouse fibroblasts to iPV neurons. The iPV neurons generated by this procedure matured 5-7 days post infection and were characterized by electrophysiological properties and known neuronal markers, such as PV and GABA. Our studies, therefore, identified an efficient approach for generating PV neurons.Parvalbumin interneurons are a small population of neurons that constitute the inhibitory modules of the cerebral cortex and hippocampus (1, 2). Although small in number and size, parvalbumin interneurons play important roles in a wide spectrum of brain functions, such as orchestrating the timing of various critical periods, synchronizing the electrical activities of certain brain cells, and rewiring the circuitry. The malfunction of these pivotal cells has been proven to be critical in autism, schizophrenia, and other neurodevelopmental disorders (1). PV 2 neurons originate primarily from the medial ganglionic eminence of the subcortical telencephalon. They mature slowly and spread throughout the brain over time. Until now the procurement and generation of specific type of neurons has depended on direct differentiation from pluripotent stem cells, such as embryonic stem cells or induced pluripotent stem cells. This common approach has been used for the generation of PV neurons (3). The generation of PV neurons for research and therapeutic purposes by other approaches needs to be investigated.Direct generation from one cell type to another provides a promising avenue for transplantable cells and holds great hope for cell-based therapy. Skin fibroblasts (mesodermal lineage) are an important source of starting cells that can be directly converted into a wide variety of cell types, including neurons (ectodermal lineage) (4, 5). Specific types of neurons, such as dopaminergic neurons (6 -8), spinal motor neurons (9), and cholinergic neurons (10) have also been efficiently converted from mouse or human fibroblasts. Thus, for late-born and slowly maturing neurons such as parvalbumin interneurons, it might be possible to surpass the laborious process of differentiation by obtaining functional cells through the direct conversion from other types of cells.Because of the side effect of integration associated with lentior retrovial delivery of exogenous genes, previous procedures used for transdifferentiation led to uncertainty in the genetic and epigenetic stability of the target cells. To explore suitable approaches for neuronal generation, we and other groups have d...

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“…Thus, iN conversion follows a hierarchical mechanism in which a sequential series of epigenetic events occurs 96 . Notably, several laboratories have shown that transcription factor expression can be temporal and is not continuously required for iN conversion, thus overcoming the potential risk that permanent overexpression of neurodevelopmental transcription factors might interfere with mature neuronal phenotypes, functionality or other relevant phenotypes of the generated iNs 97–100 (FIG. 3b,c).…”

Section: Direct Conversion To Neural Cellsmentioning

confidence: 99%

Evaluating cell reprogramming, differentiation and conversion technologies in neuroscience

et al. 2016

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The scarcity of live human brain cells for experimental access has for a long time limited our ability to study complex human neurological disorders and elucidate basic neuroscientific mechanisms. A decade ago, the development of methods to reprogramme somatic human cells into induced pluripotent stem cells enabled the in vitro generation of a wide range of neural cells from virtually any human individual. The growth of methods to generate more robust and defined neural cell types through reprogramming and direct conversion into induced neurons has led to the establishment of various human reprogramming-based neural disease models.

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Direct Neural Conversion from Human Fibroblasts Using Self-Regulating and Nonintegrating Viral Vectors

Cited by 37 publications

References 35 publications

New approaches for direct conversion of patient fibroblasts into neural cells

New approaches for direct conversion of patient fibroblasts into neural cells

Conversion of Fibroblasts to Parvalbumin Neurons by One Transcription Factor, Ascl1, and the Chemical Compound Forskolin

Evaluating cell reprogramming, differentiation and conversion technologies in neuroscience

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