title: Derivation, Characterization, and Neural Differentiation of Integration-Free Induced Pluripotent Stem Cell Lines from Parkinson's Disease Patients Carrying SNCA, LRRK2, PARK2, and GBA Mutations.
abstract: We report generation of induced pluripotent stem cell (iPSC) lines from ten Parkinson's disease (PD) patients carrying SNCA, PARK2, LRRK2, and GBA mutations, and one age-matched control. After validation of pluripotency, long-term genome stability, and integration-free reprogramming, eight of these lines (one of each SNCA, LRRK2 and GBA, four PARK2 lines, and the control) were differentiated into neural stem cells (NSC) and subsequently to dopaminergic cultures. We did not observe significant differences in the timeline of neural induction and NSC derivation between the patient and control line, nor amongst the patient lines, although we report considerable variability in the efficiency of dopaminergic differentiation among patient lines. We performed whole genome expression analyses of the lines at each stage of differentiation (fibroblast, iPSC, NSC, and dopaminergic culture) in an attempt to identify alterations by large-scale evaluation. While gene expression profiling clearly distinguished cells at different stages of differentiation, no mutation-specific clustering or difference was observed, though consistent changes in patient lines were detected in genes associated mitochondrial biology. We further examined gene expression in a stress model (MPTP-induced dopaminergic neuronal death) using two clones from the SNCA triplication line, and detected changes in genes associated with mitophagy. Our data suggested that even a well-characterized line of a monogenic disease may not be sufficient to determine the cause or mechanism of the disease, and highlights the need to use more focused strategies for large-scale data analysis.
authors.affiliation: Buck Institute for Research on Aging, Novato, CA, United States of America.
authors.forename: Olga
authors.initials: O
authors.lastname: Momcilovic
authors.affiliation: Buck Institute for Research on Aging, Novato, CA, United States of America.|University of Southern Denmark, Odense, Denmark.
authors.forename: Renuka
authors.initials: R
authors.lastname: Sivapatham
authors.affiliation: Buck Institute for Research on Aging, Novato, CA, United States of America.
authors.forename: Tal Ronnen
authors.initials: TR
authors.lastname: Oron
authors.affiliation: University of Southern Denmark, Odense, Denmark.
authors.forename: Morten
authors.initials: M
authors.lastname: Meyer
authors.affiliation: Buck Institute for Research on Aging, Novato, CA, United States of America.
authors.forename: Sean
authors.initials: S
authors.lastname: Mooney
authors.affiliation: NxCell Science, Novato, CA, United States of America.
authors.forename: Mahendra S
authors.initials: MS
authors.lastname: Rao
authors.affiliation: Buck Institute for Research on Aging, Novato, CA, United States of America.|XCell Science, Novato, CA, United States of America.
authors.forename: Xianmin
authors.initials: X
authors.lastname: Zeng
mesh_terms: D002478:Cells, Cultured; D059290:Dopaminergic Neurons / Q000166:cytology* / Q000378:metabolism; D015894:Genome, Human; D005962:Glucosylceramidase / Q000235:genetics; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 / Q000235:genetics; D008928:Mitochondria / Q000378:metabolism; D063306:Mitophagy; D009154:Mutation; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism; D055495:Neurogenesis; D010300:Parkinson Disease / Q000235:genetics* / Q000473:pathology; D044767:Ubiquitin-Protein Ligases / Q000235:genetics; D051844:alpha-Synuclein / Q000235:genetics
chemical_list: C497604:SNCA protein, human; D051844:alpha-Synuclein; D044767:Ubiquitin-Protein Ligases; C111567:parkin protein; C495280:LRRK2 protein, human; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D005962:Glucosylceramidase