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CRISPR/Cas9-Correctable mutation-related molecular and physiological phenotypes in iPSC-derived Alzheimer’s PSEN2(N141I) neurons

Basal forebrain cholinergic neurons (BFCNs) are believed to be one of the first cell types to be affected in all forms of AD, and their dysfunction is clinically correlated with impaired short-term memory formation and retrieval. We present an optimized in vitro protocol to generate human BFCNs from...

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Autores principales: Ortiz-Virumbrales, Maitane, Moreno, Cesar L., Kruglikov, Ilya, Marazuela, Paula, Sproul, Andrew, Jacob, Samson, Zimmer, Matthew, Paull, Daniel, Zhang, Bin, Schadt, Eric E., Ehrlich, Michelle E., Tanzi, Rudolph E., Arancio, Ottavio, Noggle, Scott, Gandy, Sam
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5660456/
https://www.ncbi.nlm.nih.gov/pubmed/29078805
http://dx.doi.org/10.1186/s40478-017-0475-z
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author Ortiz-Virumbrales, Maitane
Moreno, Cesar L.
Kruglikov, Ilya
Marazuela, Paula
Sproul, Andrew
Jacob, Samson
Zimmer, Matthew
Paull, Daniel
Zhang, Bin
Schadt, Eric E.
Ehrlich, Michelle E.
Tanzi, Rudolph E.
Arancio, Ottavio
Noggle, Scott
Gandy, Sam
author_facet Ortiz-Virumbrales, Maitane
Moreno, Cesar L.
Kruglikov, Ilya
Marazuela, Paula
Sproul, Andrew
Jacob, Samson
Zimmer, Matthew
Paull, Daniel
Zhang, Bin
Schadt, Eric E.
Ehrlich, Michelle E.
Tanzi, Rudolph E.
Arancio, Ottavio
Noggle, Scott
Gandy, Sam
author_sort Ortiz-Virumbrales, Maitane
collection PubMed
description Basal forebrain cholinergic neurons (BFCNs) are believed to be one of the first cell types to be affected in all forms of AD, and their dysfunction is clinically correlated with impaired short-term memory formation and retrieval. We present an optimized in vitro protocol to generate human BFCNs from iPSCs, using cell lines from presenilin 2 (PSEN2) mutation carriers and controls. As expected, cell lines harboring the PSEN2 (N141I) mutation displayed an increase in the Aβ42/40 in iPSC-derived BFCNs. Neurons derived from PSEN2 (N141I) lines generated fewer maximum number of spikes in response to a square depolarizing current injection. The height of the first action potential at rheobase current injection was also significantly decreased in PSEN2 (N141I) BFCNs. CRISPR/Cas9 correction of the PSEN2 point mutation abolished the electrophysiological deficit, restoring both the maximal number of spikes and spike height to the levels recorded in controls. Increased Aβ42/40 was also normalized following CRISPR/Cas-mediated correction of the PSEN2 (N141I) mutation. The genome editing data confirms the robust consistency of mutation-related changes in Aβ42/40 ratio while also showing a PSEN2-mutation-related alteration in electrophysiology. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article doi: (10.1186/s40478-017-0475-z) contains supplementary material, which is available to authorized users.
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spelling pubmed-56604562017-10-31 CRISPR/Cas9-Correctable mutation-related molecular and physiological phenotypes in iPSC-derived Alzheimer’s PSEN2(N141I) neurons Ortiz-Virumbrales, Maitane Moreno, Cesar L. Kruglikov, Ilya Marazuela, Paula Sproul, Andrew Jacob, Samson Zimmer, Matthew Paull, Daniel Zhang, Bin Schadt, Eric E. Ehrlich, Michelle E. Tanzi, Rudolph E. Arancio, Ottavio Noggle, Scott Gandy, Sam Acta Neuropathol Commun Research Basal forebrain cholinergic neurons (BFCNs) are believed to be one of the first cell types to be affected in all forms of AD, and their dysfunction is clinically correlated with impaired short-term memory formation and retrieval. We present an optimized in vitro protocol to generate human BFCNs from iPSCs, using cell lines from presenilin 2 (PSEN2) mutation carriers and controls. As expected, cell lines harboring the PSEN2 (N141I) mutation displayed an increase in the Aβ42/40 in iPSC-derived BFCNs. Neurons derived from PSEN2 (N141I) lines generated fewer maximum number of spikes in response to a square depolarizing current injection. The height of the first action potential at rheobase current injection was also significantly decreased in PSEN2 (N141I) BFCNs. CRISPR/Cas9 correction of the PSEN2 point mutation abolished the electrophysiological deficit, restoring both the maximal number of spikes and spike height to the levels recorded in controls. Increased Aβ42/40 was also normalized following CRISPR/Cas-mediated correction of the PSEN2 (N141I) mutation. The genome editing data confirms the robust consistency of mutation-related changes in Aβ42/40 ratio while also showing a PSEN2-mutation-related alteration in electrophysiology. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article doi: (10.1186/s40478-017-0475-z) contains supplementary material, which is available to authorized users. BioMed Central 2017-10-27 /pmc/articles/PMC5660456/ /pubmed/29078805 http://dx.doi.org/10.1186/s40478-017-0475-z Text en © The Author(s). 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Ortiz-Virumbrales, Maitane
Moreno, Cesar L.
Kruglikov, Ilya
Marazuela, Paula
Sproul, Andrew
Jacob, Samson
Zimmer, Matthew
Paull, Daniel
Zhang, Bin
Schadt, Eric E.
Ehrlich, Michelle E.
Tanzi, Rudolph E.
Arancio, Ottavio
Noggle, Scott
Gandy, Sam
CRISPR/Cas9-Correctable mutation-related molecular and physiological phenotypes in iPSC-derived Alzheimer’s PSEN2(N141I) neurons
title CRISPR/Cas9-Correctable mutation-related molecular and physiological phenotypes in iPSC-derived Alzheimer’s PSEN2(N141I) neurons
title_full CRISPR/Cas9-Correctable mutation-related molecular and physiological phenotypes in iPSC-derived Alzheimer’s PSEN2(N141I) neurons
title_fullStr CRISPR/Cas9-Correctable mutation-related molecular and physiological phenotypes in iPSC-derived Alzheimer’s PSEN2(N141I) neurons
title_full_unstemmed CRISPR/Cas9-Correctable mutation-related molecular and physiological phenotypes in iPSC-derived Alzheimer’s PSEN2(N141I) neurons
title_short CRISPR/Cas9-Correctable mutation-related molecular and physiological phenotypes in iPSC-derived Alzheimer’s PSEN2(N141I) neurons
title_sort crispr/cas9-correctable mutation-related molecular and physiological phenotypes in ipsc-derived alzheimer’s psen2(n141i) neurons
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5660456/
https://www.ncbi.nlm.nih.gov/pubmed/29078805
http://dx.doi.org/10.1186/s40478-017-0475-z
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