Reversal of Phenotypic Abnormalities by CRISPR/Cas9-Mediated Gene Correction in Huntington Disease Patient-Derived Induced Pluripotent Stem Cells

• Published in: Stem cell reports Vol. 8; no. 3; pp. 619 - 633
• Main Authors: Xu, Xiaohong, Tay, Yilin, Sim, Bernice, Yoon, Su-In, Huang, Yihui, Ooi, Jolene, Utami, Kagistia Hana, Ziaei, Amin, Ng, Bryan, Radulescu, Carola, Low, Donovan, Ng, Alvin Yu Jin, Loh, Marie, Venkatesh, Byrappa, Ginhoux, Florent, Augustine, George J., Pouladi, Mahmoud A.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 14-03-2017; Elsevier
• Subjects: Cell Differentiation - genetics; Cell Line; Cell Self Renewal - genetics; CHCHD2; CRISPR-Cas Systems; disease modeling; disease phenotypes; Electrophysiological Phenomena - genetics; Gene Editing; Gene Expression Regulation, Developmental; Gene Targeting; genetic editing; human induced pluripotent stem cell (hiPSC); Humans; Huntington disease; Huntington Disease - genetics; Huntington Disease - metabolism; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - metabolism; Mitochondria - genetics; Mitochondria - metabolism; mitochondrial dysfunction; Mitochondrial Proteins - genetics; neurodegenerative disorders; Neurons - cytology; Neurons - metabolism; Phenotype; Transcription Factors - genetics; transcriptional dysrgulation; genetic editing; CHCHD2; human induced pluripotent stem cell (hiPSC); disease phenotypes; Huntington disease; disease modeling; transcriptional dysrgulation; mitochondrial dysfunction; neurodegenerative disorders
• ISSN: 2213-6711; 2213-6711
• DOI: 10.1016/j.stemcr.2017.01.022

Huntington disease (HD) is a dominant neurodegenerative disorder caused by a CAG repeat expansion in HTT. Here we report correction of HD human induced pluripotent stem cells (hiPSCs) using a CRISPR-Cas9 and piggyBac transposon-based approach. We show that both HD and corrected isogenic hiPSCs can be differentiated into excitable, synaptically active forebrain neurons. We further demonstrate that phenotypic abnormalities in HD hiPSC-derived neural cells, including impaired neural rosette formation, increased susceptibility to growth factor withdrawal, and deficits in mitochondrial respiration, are rescued in isogenic controls. Importantly, using genome-wide expression analysis, we show that a number of apparent gene expression differences detected between HD and non-related healthy control lines are absent between HD and corrected lines, suggesting that these differences are likely related to genetic background rather than HD-specific effects. Our study demonstrates correction of HD hiPSCs and associated phenotypic abnormalities, and the importance of isogenic controls for disease modeling using hiPSCs.
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•A CRISPR-Cas9 and PiggyBac-based approach allows efficient correction of HD hiPSCs•The corrected hiPSCs can be differentiated into synaptically active neurons•Correction of HD gene mutation reverses a number of phenotypic abnormalities•Isogenic hiPSCs help distinguish mutation from genetic background-related effects
Pouladi and colleagues show that genetic correction of HD human induced pluripotent stem cells using a CRISPR-Cas9 and piggyBac transposon-based approach rescues a number of phenotypic abnormalities in derived neural cells, including increased susceptibility to growth factor withdrawal and deficits in mitochondrial respiration, and helps distinguish HD-specific from genetic background-related molecular and cellular phenotypes.