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CRISPR/Cas9 Directed Reprogramming of iPSC for Accelerated Motor Neuron Differentiation Leads to Dysregulation of Neuronal Fate Patterning and Function

Neurodegeneration causes a significant disease burden and there are few therapeutic interventions available for reversing or slowing the disease progression. Induced pluripotent stem cells (iPSCs) hold significant potential since they are sourced from adult tissue and have the capacity to be differe...

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Autores principales: Davis-Anderson, Katie, Micheva-Viteva, Sofiya, Solomon, Emilia, Hovde, Blake, Cirigliano, Elisa, Harris, Jennifer, Twary, Scott, Iyer, Rashi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10671572/
https://www.ncbi.nlm.nih.gov/pubmed/38003351
http://dx.doi.org/10.3390/ijms242216161
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author Davis-Anderson, Katie
Micheva-Viteva, Sofiya
Solomon, Emilia
Hovde, Blake
Cirigliano, Elisa
Harris, Jennifer
Twary, Scott
Iyer, Rashi
author_facet Davis-Anderson, Katie
Micheva-Viteva, Sofiya
Solomon, Emilia
Hovde, Blake
Cirigliano, Elisa
Harris, Jennifer
Twary, Scott
Iyer, Rashi
author_sort Davis-Anderson, Katie
collection PubMed
description Neurodegeneration causes a significant disease burden and there are few therapeutic interventions available for reversing or slowing the disease progression. Induced pluripotent stem cells (iPSCs) hold significant potential since they are sourced from adult tissue and have the capacity to be differentiated into numerous cell lineages, including motor neurons. This differentiation process traditionally relies on cell lineage patterning factors to be supplied in the differentiation media. Genetic engineering of iPSC with the introduction of recombinant master regulators of motor neuron (MN) differentiation has the potential to shorten and streamline cell developmental programs. We have established stable iPSC cell lines with transient induction of exogenous LHX3 and ISL1 from the Tet-activator regulatory region and have demonstrated that induction of the transgenes is not sufficient for the development of mature MNs in the absence of neuron patterning factors. Comparative global transcriptome analysis of MN development from native and Lhx-ISL1 modified iPSC cultures demonstrated that the genetic manipulation helped to streamline the neuronal patterning process. However, leaky gene expression of the exogenous MN master regulators in iPSC resulted in the premature activation of genetic pathways characteristic of the mature MN function. Dysregulation of metabolic and regulatory pathways within the developmental process affected the MN electrophysiological responses.
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spelling pubmed-106715722023-11-10 CRISPR/Cas9 Directed Reprogramming of iPSC for Accelerated Motor Neuron Differentiation Leads to Dysregulation of Neuronal Fate Patterning and Function Davis-Anderson, Katie Micheva-Viteva, Sofiya Solomon, Emilia Hovde, Blake Cirigliano, Elisa Harris, Jennifer Twary, Scott Iyer, Rashi Int J Mol Sci Article Neurodegeneration causes a significant disease burden and there are few therapeutic interventions available for reversing or slowing the disease progression. Induced pluripotent stem cells (iPSCs) hold significant potential since they are sourced from adult tissue and have the capacity to be differentiated into numerous cell lineages, including motor neurons. This differentiation process traditionally relies on cell lineage patterning factors to be supplied in the differentiation media. Genetic engineering of iPSC with the introduction of recombinant master regulators of motor neuron (MN) differentiation has the potential to shorten and streamline cell developmental programs. We have established stable iPSC cell lines with transient induction of exogenous LHX3 and ISL1 from the Tet-activator regulatory region and have demonstrated that induction of the transgenes is not sufficient for the development of mature MNs in the absence of neuron patterning factors. Comparative global transcriptome analysis of MN development from native and Lhx-ISL1 modified iPSC cultures demonstrated that the genetic manipulation helped to streamline the neuronal patterning process. However, leaky gene expression of the exogenous MN master regulators in iPSC resulted in the premature activation of genetic pathways characteristic of the mature MN function. Dysregulation of metabolic and regulatory pathways within the developmental process affected the MN electrophysiological responses. MDPI 2023-11-10 /pmc/articles/PMC10671572/ /pubmed/38003351 http://dx.doi.org/10.3390/ijms242216161 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Davis-Anderson, Katie
Micheva-Viteva, Sofiya
Solomon, Emilia
Hovde, Blake
Cirigliano, Elisa
Harris, Jennifer
Twary, Scott
Iyer, Rashi
CRISPR/Cas9 Directed Reprogramming of iPSC for Accelerated Motor Neuron Differentiation Leads to Dysregulation of Neuronal Fate Patterning and Function
title CRISPR/Cas9 Directed Reprogramming of iPSC for Accelerated Motor Neuron Differentiation Leads to Dysregulation of Neuronal Fate Patterning and Function
title_full CRISPR/Cas9 Directed Reprogramming of iPSC for Accelerated Motor Neuron Differentiation Leads to Dysregulation of Neuronal Fate Patterning and Function
title_fullStr CRISPR/Cas9 Directed Reprogramming of iPSC for Accelerated Motor Neuron Differentiation Leads to Dysregulation of Neuronal Fate Patterning and Function
title_full_unstemmed CRISPR/Cas9 Directed Reprogramming of iPSC for Accelerated Motor Neuron Differentiation Leads to Dysregulation of Neuronal Fate Patterning and Function
title_short CRISPR/Cas9 Directed Reprogramming of iPSC for Accelerated Motor Neuron Differentiation Leads to Dysregulation of Neuronal Fate Patterning and Function
title_sort crispr/cas9 directed reprogramming of ipsc for accelerated motor neuron differentiation leads to dysregulation of neuronal fate patterning and function
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10671572/
https://www.ncbi.nlm.nih.gov/pubmed/38003351
http://dx.doi.org/10.3390/ijms242216161
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