title: Reversal of Phenotypic Abnormalities by CRISPR/Cas9-Mediated Gene Correction in Huntington Disease Patient-Derived Induced Pluripotent Stem Cells.
abstract: 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.
authors.affiliation: Translational Laboratory in Genetic Medicine (TLGM), Agency for Science, Technology and Research (A(∗)STAR), 8A Biomedical Grove, Immunos, Level 5, Singapore 138648, Singapore.
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Xu
authors.affiliation: Translational Laboratory in Genetic Medicine (TLGM), Agency for Science, Technology and Research (A(∗)STAR), 8A Biomedical Grove, Immunos, Level 5, Singapore 138648, Singapore.
authors.forename: Yilin
authors.initials: Y
authors.lastname: Tay
authors.affiliation: Translational Laboratory in Genetic Medicine (TLGM), Agency for Science, Technology and Research (A(∗)STAR), 8A Biomedical Grove, Immunos, Level 5, Singapore 138648, Singapore.
authors.forename: Bernice
authors.initials: B
authors.lastname: Sim
authors.affiliation: Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore 637553, Singapore.
authors.forename: Su-In
authors.initials: SI
authors.lastname: Yoon
authors.affiliation: Translational Laboratory in Genetic Medicine (TLGM), Agency for Science, Technology and Research (A(∗)STAR), 8A Biomedical Grove, Immunos, Level 5, Singapore 138648, Singapore.
authors.forename: Yihui
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Translational Laboratory in Genetic Medicine (TLGM), Agency for Science, Technology and Research (A(∗)STAR), 8A Biomedical Grove, Immunos, Level 5, Singapore 138648, Singapore.
authors.forename: Jolene
authors.initials: J
authors.lastname: Ooi
authors.affiliation: Translational Laboratory in Genetic Medicine (TLGM), Agency for Science, Technology and Research (A(∗)STAR), 8A Biomedical Grove, Immunos, Level 5, Singapore 138648, Singapore.
authors.forename: Kagistia Hana
authors.initials: KH
authors.lastname: Utami
authors.affiliation: Translational Laboratory in Genetic Medicine (TLGM), Agency for Science, Technology and Research (A(∗)STAR), 8A Biomedical Grove, Immunos, Level 5, Singapore 138648, Singapore.
authors.forename: Amin
authors.initials: A
authors.lastname: Ziaei
authors.affiliation: Translational Laboratory in Genetic Medicine (TLGM), Agency for Science, Technology and Research (A(∗)STAR), 8A Biomedical Grove, Immunos, Level 5, Singapore 138648, Singapore.
authors.forename: Bryan
authors.initials: B
authors.lastname: Ng
authors.affiliation: Translational Laboratory in Genetic Medicine (TLGM), Agency for Science, Technology and Research (A(∗)STAR), 8A Biomedical Grove, Immunos, Level 5, Singapore 138648, Singapore.
authors.forename: Carola
authors.initials: C
authors.lastname: Radulescu
authors.affiliation: Singapore Immunology Network (SIgN), A(∗)STAR, Singapore 138648, Singapore.
authors.forename: Donovan
authors.initials: D
authors.lastname: Low
authors.affiliation: Comparative Genomics Laboratory, Institute of Molecular and Cell Biology, A(∗)STAR, Biopolis, Singapore 138673, Singapore.
authors.forename: Alvin Yu Jin
authors.initials: AYJ
authors.lastname: Ng
authors.affiliation: Translational Laboratory in Genetic Medicine (TLGM), Agency for Science, Technology and Research (A(∗)STAR), 8A Biomedical Grove, Immunos, Level 5, Singapore 138648, Singapore.
authors.forename: Marie
authors.initials: M
authors.lastname: Loh
authors.affiliation: Comparative Genomics Laboratory, Institute of Molecular and Cell Biology, A(∗)STAR, Biopolis, Singapore 138673, Singapore; Department of Paediatrics, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 119228, Singapore.
authors.forename: Byrappa
authors.initials: B
authors.lastname: Venkatesh
authors.affiliation: Singapore Immunology Network (SIgN), A(∗)STAR, Singapore 138648, Singapore.
authors.forename: Florent
authors.initials: F
authors.lastname: Ginhoux
authors.affiliation: Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore 637553, Singapore; Institute of Molecular and Cell Biology (IMCB), Singapore 138673, Singapore.
authors.forename: George J
authors.initials: GJ
authors.lastname: Augustine
authors.affiliation: Translational Laboratory in Genetic Medicine (TLGM), Agency for Science, Technology and Research (A(∗)STAR), 8A Biomedical Grove, Immunos, Level 5, Singapore 138648, Singapore; Department of Medicine, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597, Singapore. Electronic address: map@pouladilab.org.
authors.forename: Mahmoud A
authors.initials: MA
authors.lastname: Pouladi
mesh_terms: D064113:CRISPR-Cas Systems; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D000066673:Cell Self Renewal / Q000235:genetics; D004268:DNA-Binding Proteins; D055724:Electrophysiological Phenomena / Q000235:genetics; D000072669:Gene Editing; D018507:Gene Expression Regulation, Developmental; D018390:Gene Targeting; D006801:Humans; D006816:Huntington Disease / Q000235:genetics* / Q000378:metabolism*; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D008928:Mitochondria / Q000235:genetics / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000235:genetics; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D010641:Phenotype; D014157:Transcription Factors / Q000235:genetics
chemical_list: C580880:CHCHD2 protein, human; D004268:DNA-Binding Proteins; D024101:Mitochondrial Proteins; D014157:Transcription Factors
keywords: CHCHD2; Huntington disease; disease modeling; disease phenotypes; genetic editing; human induced pluripotent stem cell (hiPSC); mitochondrial dysfunction; neurodegenerative disorders; transcriptional dysrgulation