title: Gene expression signatures in motor neurone disease fibroblasts reveal dysregulation of metabolism, hypoxia-response and RNA processing functions.
abstract: OBJECTIVE
  Amyotrophic lateral sclerosis (ALS) and primary lateral sclerosis (PLS) are two syndromic variants within the motor neurone disease spectrum. As PLS and most ALS cases are sporadic (SALS), this limits the availability of cellular models for investigating pathogenic mechanisms and therapeutic targets. The aim of this study was to use gene expression profiling to evaluate fibroblasts as cellular models for SALS and PLS, to establish whether dysregulated biological processes recapitulate those seen in the central nervous system and to elucidate pathways that distinguish the clinically defined variants of SALS and PLS.
  
  
  METHODS
  Microarray analysis was performed on fibroblast RNA and differentially expressed genes identified. Genes in enriched biological pathways were validated by quantitative PCR and functional assays performed to establish the effect of altered RNA levels on the cellular processes.
  
  
  RESULTS
  Gene expression profiling demonstrated that whilst there were many differentially expressed genes in common between SALS and PLS fibroblasts, there were many more expressed specifically in the SALS fibroblasts, including those involved in RNA processing and the stress response. Functional analysis of the fibroblasts confirmed a significant decrease in miRNA production and a reduced response to hypoxia in SALS fibroblasts. Furthermore, metabolic gene changes seen in SALS, many of which were also evident in PLS fibroblasts, resulted in dysfunctional cellular respiration.
  
  
  CONCLUSIONS
  The data demonstrate that fibroblasts can act as cellular models for ALS and PLS, by establishing the transcriptional changes in known pathogenic pathways that confer subsequent functional effects and potentially highlight targets for therapeutic intervention.
authors.affiliation: Sheffield Institute for Translational Neuroscience (SITraN), Department of Neuroscience, University of Sheffield, Sheffield, UK.
authors.forename: Rohini
authors.initials: R
authors.lastname: Raman
authors.forename: Scott P
authors.initials: SP
authors.lastname: Allen
authors.forename: Emily F
authors.initials: EF
authors.lastname: Goodall
authors.forename: Shelley
authors.initials: S
authors.lastname: Kramer
authors.forename: Lize-Linde
authors.initials: LL
authors.lastname: Ponger
authors.forename: Paul R
authors.initials: PR
authors.lastname: Heath
authors.forename: Marta
authors.initials: M
authors.lastname: Milo
authors.forename: Hannah C
authors.initials: HC
authors.lastname: Hollinger
authors.forename: Theresa
authors.initials: T
authors.lastname: Walsh
authors.forename: J Robin
authors.initials: JR
authors.lastname: Highley
authors.forename: Simon
authors.initials: S
authors.lastname: Olpin
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: McDermott
authors.forename: Pamela J
authors.initials: PJ
authors.lastname: Shaw
authors.forename: Janine
authors.initials: J
authors.lastname: Kirby
mesh_terms: D000328:Adult; D000368:Aged; D015687:Cell Hypoxia / Q000502:physiology; D002478:Cells, Cultured; D005260:Female; D005347:Fibroblasts / Q000378:metabolism* / Q000473:pathology*; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D015151:Immunoblotting; D008297:Male; D035683:MicroRNAs / Q000032:analysis; D008875:Middle Aged; D016472:Motor Neuron Disease / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D059467:Transcriptome
chemical_list: D035683:MicroRNAs
keywords: amyotrophic lateral sclerosis; cell models; fibroblasts; hypoxia response; microRNA; microarray; primary lateral sclerosis
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