title: NRF2 activation restores disease related metabolic deficiencies in olfactory neurosphere-derived cells from patients with sporadic Parkinson's disease.
abstract: BACKGROUND
  Without appropriate cellular models the etiology of idiopathic Parkinson's disease remains unknown. We recently reported a novel patient-derived cellular model generated from biopsies of the olfactory mucosa (termed olfactory neurosphere-derived (hONS) cells) which express functional and genetic differences in a disease-specific manner. Transcriptomic analysis of Patient and Control hONS cells identified the NRF2 transcription factor signalling pathway as the most differentially expressed in Parkinson's disease.
  
  
  RESULTS
  We tested the robustness of our initial findings by including additional cell lines and confirmed that hONS cells from Patients had 20% reductions in reduced glutathione levels and MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt] metabolism compared to cultures from healthy Control donors. We also confirmed that Patient hONS cells are in a state of oxidative stress due to higher production of H(2)O(2) than Control cultures. siRNA-mediated ablation of NRF2 in Control donor cells decreased both total glutathione content and MTS metabolism to levels detected in cells from Parkinson's Disease patients. Conversely, and more importantly, we showed that activation of the NRF2 pathway in Parkinson's disease hONS cultures restored glutathione levels and MTS metabolism to Control levels. Paradoxically, transcriptomic analysis after NRF2 pathway activation revealed an increased number of differentially expressed mRNAs within the NRF2 pathway in L-SUL treated Patient-derived hONS cells compared to L-SUL treated Controls, even though their metabolism was restored to normal. We also identified differential expression of the PI3K/AKT signalling pathway, but only post-treatment.
  
  
  CONCLUSIONS
  Our results confirmed NRF2 as a potential therapeutic target for Parkinson's disease and provided the first demonstration that NRF2 function was inducible in Patient-derived cells from donors with uniquely varied genetic backgrounds. However, our results also demonstrated that the response of PD patient-derived cells was not co-ordinated in the same way as in Control cells. This may be an important factor when developing new therapeutics.
authors: National Centre for Adult Stem Cell Research, Eskitis Institute for Cell and Molecular Therapies, Griffith University, Brisbane, Queensland, Australia.
authors: Anthony L
authors: AL
authors: Cook
authors: Alejandra M
authors: AM
authors: Vitale
authors: Sugandha
authors: S
authors: Ravishankar
authors: Nicholas
authors: N
authors: Matigian
authors: Greg T
authors: GT
authors: Sutherland
authors: Jiangou
authors: J
authors: Shan
authors: Ratneswary
authors: R
authors: Sutharsan
authors: Chris
authors: C
authors: Perry
authors: Peter A
authors: PA
authors: Silburn
authors: George D
authors: GD
authors: Mellick
authors: Murray L
authors: ML
authors: Whitelaw
authors: Christine A
authors: CA
authors: Wells
authors: Alan
authors: A
authors: Mackay-Sim
authors: Stephen A
authors: SA
authors: Wood
mesh_terms: D016022:Case-Control Studies; D002460:Cell Line; D005260:Female; D020868:Gene Silencing; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D017879:Isothiocyanates; D008297:Male; D051267:NF-E2-Related Factor 2 / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D009831:Olfactory Mucosa / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology*; D010300:Parkinson Disease / Q000378:metabolism* / Q000473:pathology*; D015398:Signal Transduction / Q000187:drug effects; D013454:Sulfoxides; D013778:Tetrazolium Salts / Q000378:metabolism; D013844:Thiazoles / Q000378:metabolism; D013861:Thiocyanates / Q000494:pharmacology
chemical_list: D017879:Isothiocyanates; D051267:NF-E2-Related Factor 2; D013454:Sulfoxides; D013778:Tetrazolium Salts; D013844:Thiazoles; D013861:Thiocyanates; C070380:3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium; C016766:sulforaphane; D005978:Glutathione