Activities per year
‘nutrient overload' NAFLD model in human hepatic C3A cells, to recapitulate the sequence of events
proposed to occur in NAFLD.
Objectives: To develop a systems biology approach (HepSys) combining identification of distinct patterns
of perturbed hepatic metabolism, with downstream integration of proteomics and metabolomics data sets.
Methods: C3A cells were loaded with a 'nutrient cocktail' (72h) of Lactate, Pyruvate, Octanoate and
Ammonia (LPON) to promote the dysregulated metabolic NAFLD phenotype. Microarray RNA expression
was measured using Illumina® Whole Human Genome BeadChip H12 Microarray. Data was normalized,
validated and analysed to examine differences between Control (C3A untreated) and LPON-treated
Results: NAFLD phenotype was confirmed using morphological [BODIPY] and biochemical parameters
[MitoSOX]. Microarray data was collated onto the HepSys Microarray Portal with capacity for Proteomics
data integration. 12 distinct biochemical pathways were identified [Control v LPON] with significantly
upregulated differential expression patterns [Filters/ Statistics: >2x fold Deltaexpression; P<0.05, Rank
Products pfp; and linked to KEGG Metabolic Pathway]; including: Metabolic pathways [30 genes; eg
FASN, involved in fatty acid metabolism, ACAT2 (synthesizes Acetoacetyl CoA, essential for cholesterol
synthesis; PCK2 key mitochondrial enzyme in gluconeogenesis; Steroid biosynthesis [5 genes: MSMO1,
TM7SF2 FDFT1, DHCR7, EBP (EBP is involved in cholesterol biosynthesis/ lipoprotein internalisation];
Oxidative phosphorylation [6 genes total: eg NADH dehydrogenases: NDUFA3, NDUFB10, NDUFB7;
Cytochrome c reductase: CYC1; ATP6V0E2 (H+-ATPase) and ATP5D (catalyzes ATP synthesis). Global
changes in expression levels were visualized using Heatmap [138 log2 expression values, samples
clustered via complete linkage].
Conclusions: Our results show NAFLD-related morphological/ biochemical changes are associated with
clear differential gene expression patterns, which implicate several distinct pathways in the dysregulated
metabolism of NAFLD. Future validatory studies of gene expression signatures, including qRT-PCR
analyses, and correlation/ integration with cognate Proteomic data sets may uncover hitherto unknown
pathways involved in NAFLD pathogenesis.
|Publication status||Published - Feb 2013|
|Event||lEASL Monothematic Conference: Systems Biology of the Liver: Systems Biology and Clinics Face-à-Face - Luxemburg, Luxembourg|
Duration: 21 Feb 2013 → 23 Feb 2013
|Conference||lEASL Monothematic Conference: Systems Biology of the Liver: Systems Biology and Clinics Face-à-Face|
|Period||21/02/13 → 23/02/13|
FingerprintDive into the research topics of 'HEPSYS-I MICROARRAY EXPRESSION PROFILING OF AN IN VITRO 'NUTRIENT EXCESS' MODEL OF NAFLD IN HUMAN HEPATOCYTES'. Together they form a unique fingerprint.
- 1 Participation in conference
Philipp Treskes (Participant)21 Feb 2013 → 23 Feb 2013
Activity: Participating in or organising an event types › Participation in conference