title: LRRK2 mutations cause mitochondrial DNA damage in iPSC-derived neural cells from Parkinson's disease patients: reversal by gene correction.
abstract: Parkinson's disease associated mutations in leucine rich repeat kinase 2 (LRRK2) impair mitochondrial function and increase the vulnerability of induced pluripotent stem cell (iPSC)-derived neural cells from patients to oxidative stress. Since mitochondrial DNA (mtDNA) damage can compromise mitochondrial function, we examined whether LRRK2 mutations can induce damage to the mitochondrial genome. We found greater levels of mtDNA damage in iPSC-derived neural cells from patients carrying homozygous or heterozygous LRRK2 G2019S mutations, or at-risk individuals carrying the heterozygous LRRK2 R1441C mutation, than in cells from unrelated healthy subjects who do not carry LRRK2 mutations. After zinc finger nuclease-mediated repair of the LRRK2 G2019S mutation in iPSCs, mtDNA damage was no longer detected in differentiated neuroprogenitor and neural cells. Our results unambiguously link LRRK2 mutations to mtDNA damage and validate a new cellular phenotype that can be used for examining pathogenic mechanisms and screening therapeutic strategies.
authors.affiliation: Pittsburgh Institute for Neurodegenerative Diseases, Department of Neurology, University of Pittsburgh, Pittsburgh, PA 15260, USA.
authors.forename: Laurie H
authors.initials: LH
authors.lastname: Sanders
authors.affiliation: Sangamo BioSciences, Inc., Point Richmond Tech Center, 501 Canal Boulevard, Suite A100, Richmond, CA 94804, USA.
authors.forename: Josée
authors.initials: J
authors.lastname: Laganière
authors.affiliation: Neuroregeneration Institute, McLean Hospital/Harvard Medical School, Belmont, MA 02478, USA.
authors.forename: Oliver
authors.initials: O
authors.lastname: Cooper
authors.affiliation: The Parkinson's Institute, 675 Almanor Avenue, Sunnyvale, CA 94025, USA.
authors.forename: Sally K
authors.initials: SK
authors.lastname: Mak
authors.affiliation: Sangamo BioSciences, Inc., Point Richmond Tech Center, 501 Canal Boulevard, Suite A100, Richmond, CA 94804, USA.
authors.forename: B Joseph
authors.initials: BJ
authors.lastname: Vu
authors.affiliation: The Parkinson's Institute, 675 Almanor Avenue, Sunnyvale, CA 94025, USA.
authors.forename: Y Anne
authors.initials: YA
authors.lastname: Huang
authors.affiliation: Sangamo BioSciences, Inc., Point Richmond Tech Center, 501 Canal Boulevard, Suite A100, Richmond, CA 94804, USA.
authors.forename: David E
authors.initials: DE
authors.lastname: Paschon
authors.affiliation: The Parkinson's Institute, 675 Almanor Avenue, Sunnyvale, CA 94025, USA.
authors.forename: Malini
authors.initials: M
authors.lastname: Vangipuram
authors.affiliation: The Parkinson's Institute, 675 Almanor Avenue, Sunnyvale, CA 94025, USA.
authors.forename: Ramya
authors.initials: R
authors.lastname: Sundararajan
authors.affiliation: Sangamo BioSciences, Inc., Point Richmond Tech Center, 501 Canal Boulevard, Suite A100, Richmond, CA 94804, USA.
authors.forename: Fyodor D
authors.initials: FD
authors.lastname: Urnov
authors.affiliation: The Parkinson's Institute, 675 Almanor Avenue, Sunnyvale, CA 94025, USA.
authors.forename: J William
authors.initials: JW
authors.lastname: Langston
authors.affiliation: Sangamo BioSciences, Inc., Point Richmond Tech Center, 501 Canal Boulevard, Suite A100, Richmond, CA 94804, USA.
authors.forename: Philip D
authors.initials: PD
authors.lastname: Gregory
authors.affiliation: Sangamo BioSciences, Inc., Point Richmond Tech Center, 501 Canal Boulevard, Suite A100, Richmond, CA 94804, USA.
authors.forename: H Steve
authors.initials: HS
authors.lastname: Zhang
authors.affiliation: Pittsburgh Institute for Neurodegenerative Diseases, Department of Neurology, University of Pittsburgh, Pittsburgh, PA 15260, USA. Electronic address: jgreena@pitt.edu.
authors.forename: J Timothy
authors.initials: JT
authors.lastname: Greenamyre
authors.affiliation: Neuroregeneration Institute, McLean Hospital/Harvard Medical School, Belmont, MA 02478, USA. Electronic address: isacson@hms.harvard.edu.
authors.forename: Ole
authors.initials: O
authors.lastname: Isacson
authors.affiliation: The Parkinson's Institute, 675 Almanor Avenue, Sunnyvale, CA 94025, USA. Electronic address: bschuele@thepi.org.
authors.forename: Birgitt
authors.initials: B
authors.lastname: Schüle
mesh_terms: D000328:Adult; D000368:Aged; D004249:DNA Damage; D004260:DNA Repair; D004272:DNA, Mitochondrial / Q000235:genetics / Q000378:metabolism*; D005260:Female; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D008297:Male; D008875:Middle Aged; D009154:Mutation; D058953:Neural Stem Cells / Q000378:metabolism*; D010300:Parkinson Disease / Q000235:genetics* / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics*; D052416:Targeted Gene Repair; D016335:Zinc Fingers
chemical_list: D004272:DNA, Mitochondrial; C495280:LRRK2 protein, human; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D017346:Protein Serine-Threonine Kinases
keywords: LRRK2; Mitochondrial DNA damage; Parkinson's disease; Stem cells