title: Necrosis is increased in lymphoblastoid cell lines from children with autism compared with their non-autistic siblings under conditions of oxidative and nitrosative stress.
abstract: Autism spectrum disorders are a heterogeneous group of neurodevelopmental conditions characterised by impairments in reciprocal social interaction, communication and stereotyped behaviours. As increased DNA damage events have been observed in a range of other neurological disorders, it was hypothesised that they would be elevated in lymphoblastoid cell lines (LCLs) obtained from children with autism compared with their non-autistic siblings. Six case-sibling pairs of LCLs from children with autistic disorder and their non-autistic siblings were obtained from the Autism Genetic Resource Exchange (AGRE) and cultured in standard RPMI-1640 tissue culture medium. Cells were exposed to medium containing either 0, 25, 50, 100 and 200 µM hydrogen peroxide (an oxidative stressor) or 0, 5, 10, 20 and 40 µM s-nitroprusside (a nitric oxide producer) for 1h. Following exposure, the cells were microscopically scored for DNA damage, cytostasis and cytotoxicity biomarkers as measured using the cytokinesis-block micronucleus cytome assay. Necrosis was significantly increased in cases relative to controls when exposed to oxidative and nitrosative stress (P = 0.001 and 0.01, respectively). Nuclear division index was significantly lower in LCLs from children with autistic disorder than their non-autistic siblings when exposed to hydrogen peroxide (P = 0.016), but there was no difference in apoptosis, micronucleus frequency, nucleoplasmic bridges or nuclear buds. Exposure to s-nitroprusside significantly increased the number of micronuclei in non-autistic siblings compared with cases (P = 0.003); however, other DNA damage biomarkers, apoptosis and nuclear division did not differ significantly between groups. The findings of this study show (i) that LCLs from children with autism are more sensitive to necrosis under conditions of oxidative and nitrosative stress than their non-autistic siblings and (ii) refutes the hypothesis that children with autistic disorder are abnormally susceptible to DNA damage.
authors.affiliation: Sansom Institute of Health Research, University of South Australia, North Terrace, Adelaide, Australia.
authors.forename: Penelope A E
authors.initials: PA
authors.lastname: Main
authors.forename: Philip
authors.initials: P
authors.lastname: Thomas
authors.forename: Adrian
authors.initials: A
authors.lastname: Esterman
authors.forename: Michael F
authors.initials: MF
authors.lastname: Fenech
mesh_terms: D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D001321:Autistic Disorder / Q000235:genetics* / Q000378:metabolism*; D002460:Cell Line; D002648:Child; D002659:Child Development Disorders, Pervasive / Q000235:genetics / Q000378:metabolism; D002675:Child, Preschool; D004249:DNA Damage / Q000187:drug effects / Q000235:genetics; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008214:Lymphocytes / Q000378:metabolism; D008297:Male; D009336:Necrosis / Q000235:genetics* / Q000378:metabolism*; D009599:Nitroprusside / Q000494:pharmacology; D015538:Nitrosation; D010084:Oxidation-Reduction; D035781:Siblings; D013312:Stress, Physiological
chemical_list: D009599:Nitroprusside; D006861:Hydrogen Peroxide