Engineering synucleinopathy-resistant human dopaminergic neurons by CRISPR-mediated deletion of the SNCA gene.

Tytuł:: Engineering synucleinopathy-resistant human dopaminergic neurons by CRISPR-mediated deletion of the SNCA gene.
Autorzy:: Chen Y; MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, School of Biological Sciences, The University of Edinburgh, Edinburgh, UK.; UK Centre for Mammalian Synthetic Biology, The University of Edinburgh, Edinburgh, UK.
Dolt KS; MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, School of Biological Sciences, The University of Edinburgh, Edinburgh, UK.
Kriek M; UCB Pharma Ltd., Slough, UK.
Baker T; UCB Pharma Ltd., Slough, UK.
Downey P; UCB BioPharma Sprl., Braine-l'Alleud, Belgium.
Drummond NJ; MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, School of Biological Sciences, The University of Edinburgh, Edinburgh, UK.
Canham MA; MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, School of Biological Sciences, The University of Edinburgh, Edinburgh, UK.
Natalwala A; MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, School of Biological Sciences, The University of Edinburgh, Edinburgh, UK.
Rosser S; UK Centre for Mammalian Synthetic Biology, The University of Edinburgh, Edinburgh, UK.
Kunath T; MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, School of Biological Sciences, The University of Edinburgh, Edinburgh, UK.; UK Centre for Mammalian Synthetic Biology, The University of Edinburgh, Edinburgh, UK.
Źródło:: The European journal of neuroscience [Eur J Neurosci] 2019 Feb; Vol. 49 (4), pp. 510-524. Date of Electronic Publication: 2018 Dec 21.
Typ publikacji:: Journal Article; Research Support, Non-U.S. Gov't
Język:: English
Imprint Name(s):: Publication: : Oxford : Wiley-Blackwell
Original Publication: Oxford, UK : Published on behalf of the European Neuroscience Association by Oxford University Press, c1989-
MeSH Terms:: CRISPR-Associated Protein 9*
Cell Differentiation*
Dopaminergic Neurons*
Embryonic Stem Cells*
Gene Editing*
Synucleinopathies*
alpha-Synuclein*
Parkinson Disease/*therapy
Cell Line ; Humans ; Mesencephalon/cytology
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Grant Information:: BB/M018040/1 United Kingdom BB_ Biotechnology and Biological Sciences Research Council; F-0902 United Kingdom PUK_ Parkinson's UK; MR/K017276/1 United Kingdom MRC_ Medical Research Council; United Kingdom WT_ Wellcome Trust; MR/J012831/1 United Kingdom MRC_ Medical Research Council; 203646/Z/16/Z United Kingdom WT_ Wellcome Trust
Contributed Indexing:: Keywords: CRISPR/Cas9n; disease-resistant; dopaminergic neurons; hESCs; synucleinopathy
Substance Nomenclature:: 0 (SNCA protein, human)
0 (alpha-Synuclein)
EC 3.1.- (CRISPR-Associated Protein 9)
Entry Date(s):: Date Created: 20181126 Date Completed: 20200622 Latest Revision: 20240214
Update Code:: 20240214
PubMed Central ID:: PMC6492083
DOI:: 10.1111/ejn.14286
PMID:: 30472757
: Czasopismo naukowe

Pełny tekst

An emerging treatment for Parkinson's disease (PD) is cell replacement therapy. Authentic midbrain dopaminergic (mDA) neuronal precursors can be differentiated from human embryonic stem cells (hESCs) and human induced pluripotent stem cells (iPSCs). These laboratory-generated mDA cells have been demonstrated to mature into functional dopaminergic neurons upon transplantation into preclinical models of PD. However, clinical trials with human fetal mesenchephalic cells have shown that cell replacement grafts in PD are susceptible to Lewy body formation suggesting host-to-graft transfer of α-synuclein pathology. Here, we have used CRISPR/Cas9n technology to delete the endogenous SNCA gene, encoding for α-synuclein, in a clinical-grade hESC line to generate SNCA +/- and SNCA -/- cell lines. These hESC lines were first differentiated into mDA neurons, and then challenged with recombinant α-synuclein preformed fibrils (PFFs) to seed the formation for Lewy-like pathology as measured by phosphorylation of serine-129 of α-synuclein (pS129-αSyn). Wild-type neurons were fully susceptible to the formation of protein aggregates positive for pS129-αSyn, while SNCA +/- and SNCA -/- neurons exhibited significant resistance to the formation of this pathological mark. This work demonstrates that reducing or completely removing SNCA alleles by CRISPR/Cas9n-mediated gene editing confers a measure of resistance to Lewy pathology.
(© 2018 The Authors. European Journal of Neuroscience published by Federation of European Neuroscience Societies and John Wiley & Sons Ltd.)

Comment in: Eur J Neurosci. 2019 Feb;49(3):316-319. (PMID: 30614081)

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