JoussefHayek and Giuseppe Valacchi contributed equally to this work.Abbreviations: 4-HNE, 4-hydroxynonenal; ASD, autism spectrum disorder; ATP5A, ATP synthase subunit alpha, mitochondrial; COX4, cytochrome c oxidase subunit 4 isoform 1, mitochondrial; DMEM, Dulbecco's Modification of Eagle's Medium; DRP1, dynamin-1-like protein; ECAR, extracellular acidification rate; ETC, electron transport chain; FBS, fetal bovine serum; FCCP, carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone; FIS1, mitochondrial fission 1 protein; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; GSH, glutathione; H 2 O 2 , hydrogen peroxide; HBSS, Hanks' balanced salt solution; HDCA1, histone deacetylase 1; HDL, high-density lipoproteins; HO-1, heme oxygenase 1; MFN1, mitofusin-1; MFN2, mitofusin-2; NADPH, nicotinamide adenine dinucleotide phosphate; NDUFB8, NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 8, mitochondrial; NFκB, nuclear factor NF-kappa-B; NOX, NADPH (Nicotinamide adenine dinucleotide phosphate) oxidase; NPBI, nonprotein-bound iron; NQO1, NAD(P)H quinone dehydrogenase 1; Nrf2, nuclear factor erythroid 2-related factor 2; OCR, oxygen consuming ratio; OPA1, dynamin-like 120 kDa protein, mitochondrial; Parkin, E3 ubiquitin-protein ligase parkin; PBMC, peripheral blood mononuclear cells; PBS, phosphate-buffered saline; PINK1, serine/threonine-protein kinase PINK1, mitochondrial; PMA, phorbol myristate acetate; PON-1, paraoxonase-1; PVDF, polyvinylidene difluoride; RFU, relative fluorescence units; RIPA, radio-immunoprecipitation assay; RLU, relative luminescence units; ROS, reactive oxygen species; SDHA, succinate dehydrogenase complex flavoprotein subunit A; SIRT3, sirtuin 3; SOD, superoxide dismutase; TBS-T, tris-buffered saline, 0.1% Tween 20; TEM, transmission electron microscopy; TOMM20, translocase of outer mitochondrial membrane 20; TRX, thioredoxin; TXNIP, thioredoxin-interacting protein; UCP2, mitochondrial uncoupling protein 2; UQCRC2, cytochrome b-c1 complex subunit 2, mitochondrial.
AbstractAutism spectrum disorder (ASD) has been hypothesized to be a result of the interplay between genetic predisposition and increased vulnerability to early environmental insults. Mitochondrial dysfunctions appear also involved in ASD pathophysiology, but the mechanisms by which such alterations develop are not completely understood.Here, we analyzed ASD primary fibroblasts by measuring mitochondrial bioenergetics, ultrastructural and dynamic parameters to investigate the hypothesis that defects in these pathways could be interconnected phenomena responsible or consequence for the redox imbalance observed in ASD. High levels of 4-hydroxynonenal protein adducts together with increased NADPH (nicotinamide adenine dinucleotide phosphateoxidase) activity and mitochondrial superoxide production coupled with a compromised antioxidant response guided by a defective Nuclear Factor Erythroid 2-Related Factor 2 pathway confirmed an unbalanced redox homeostasis in ASD.Moreover, ASD fibroblasts showed overactive mitochondrial bioenerget...