title: Genomic Instability Associated with p53 Knockdown in the Generation of Huntington's Disease Human Induced Pluripotent Stem Cells.
abstract: Alterations in DNA damage response and repair have been observed in Huntington's disease (HD). We generated induced pluripotent stem cells (iPSC) from primary dermal fibroblasts of 5 patients with HD and 5 control subjects. A significant fraction of the HD iPSC lines had genomic abnormalities as assessed by karyotype analysis, while none of our control lines had detectable genomic abnormalities. We demonstrate a statistically significant increase in genomic instability in HD cells during reprogramming. We also report a significant association with repeat length and severity of this instability. Our karyotypically normal HD iPSCs also have elevated ATM-p53 signaling as shown by elevated levels of phosphorylated p53 and H2AX, indicating either elevated DNA damage or hypersensitive DNA damage signaling in HD iPSCs. Thus, increased DNA damage responses in the HD genotype is coincidental with the observed chromosomal aberrations. We conclude that the disease causing mutation in HD increases the propensity of chromosomal instability relative to control fibroblasts specifically during reprogramming to a pluripotent state by a commonly used episomal-based method that includes p53 knockdown.
authors.affiliation: Department of Neurology, Vanderbilt University Medical Center, Nashville, TN, 37240, United States of America.|Vanderbilt Brain Institute, Vanderbilt University, Nashville, TN, 37232, United States of America.
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Tidball
authors.affiliation: Department of Neurology, Vanderbilt University Medical Center, Nashville, TN, 37240, United States of America.|Vanderbilt Brain Institute, Vanderbilt University, Nashville, TN, 37232, United States of America.
authors.forename: M Diana
authors.initials: MD
authors.lastname: Neely
authors.affiliation: Genetics Associates Inc., Nashville, TN, 37203, United States of America.
authors.forename: Reed
authors.initials: R
authors.lastname: Chamberlin
authors.affiliation: Department of Neurology, Vanderbilt University Medical Center, Nashville, TN, 37240, United States of America.
authors.forename: Asad A
authors.initials: AA
authors.lastname: Aboud
authors.affiliation: Department of Neurology, Vanderbilt University Medical Center, Nashville, TN, 37240, United States of America.|Vanderbilt Brain Institute, Vanderbilt University, Nashville, TN, 37232, United States of America.
authors.forename: Kevin K
authors.initials: KK
authors.lastname: Kumar
authors.affiliation: Department of Neurology, Vanderbilt University Medical Center, Nashville, TN, 37240, United States of America.
authors.forename: Bingying
authors.initials: B
authors.lastname: Han
authors.affiliation: Department of Neurology, Vanderbilt University Medical Center, Nashville, TN, 37240, United States of America.|Vanderbilt Brain Institute, Vanderbilt University, Nashville, TN, 37232, United States of America.
authors.forename: Miles R
authors.initials: MR
authors.lastname: Bryan
authors.affiliation: Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, NY, 10461, United States of America.
authors.forename: Michael
authors.initials: M
authors.lastname: Aschner
authors.affiliation: Vanderbilt Brain Institute, Vanderbilt University, Nashville, TN, 37232, United States of America.|Department of Pediatrics, Vanderbilt University Medical Center, Nashville, TN, 37240, United States of America.
authors.forename: Kevin C
authors.initials: KC
authors.lastname: Ess
authors.affiliation: Department of Neurology, Vanderbilt University Medical Center, Nashville, TN, 37240, United States of America.|Vanderbilt Brain Institute, Vanderbilt University, Nashville, TN, 37232, United States of America.|Center in Molecular Toxicology, Vanderbilt University, Nashville, TN, 37232, United States of America.
authors.forename: Aaron B
authors.initials: AB
authors.lastname: Bowman
mesh_terms: D000328:Adult; D000368:Aged; D002478:Cells, Cultured; D004249:DNA Damage; D055785:Gene Knockdown Techniques; D042822:Genomic Instability; D006801:Humans; D006816:Huntington Disease / Q000235:genetics / Q000473:pathology*; D057026:Induced Pluripotent Stem Cells / Q000473:pathology*; D007621:Karyotyping; D008875:Middle Aged; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects; D016159:Tumor Suppressor Protein p53 / Q000235:genetics*; D055815:Young Adult; D009353:Zinostatin / Q000494:pharmacology
chemical_list: D019384:Nucleic Acid Synthesis Inhibitors; D016159:Tumor Suppressor Protein p53; D009353:Zinostatin