title: Tryptophan-NAD+ pathway metabolites as putative biomarkers and predictors of peroxisome proliferation.
abstract: The present study was designed to provide further information about the relevance of raised urinary levels of N-methylnicotinamide (NMN), and/or its metabolites N-methyl-4-pyridone-3-carboxamide (4PY) and N-methyl-2-pyridone-3-carboxamide (2PY), to peroxisome proliferation by dosing rats with known peroxisome proliferator-activated receptor alpha (PPARalpha) ligands [fenofibrate, diethylhexylphthalate (DEHP) and long-chain fatty acids (LCFA)] and other compounds believed to modulate lipid metabolism via PPARalpha-independent mechanisms (simvastatin, hydrazine and chlorpromazine). Urinary NMN was correlated with standard markers of peroxisome proliferation and serum lipid parameters with the aim of establishing whether urinary NMN could be used as a biomarker for peroxisome proliferation in the rat. Data from this study were also used to validate a previously constructed multivariate statistical model of peroxisome proliferation (PP) in the rat. The predictive model, based on 1H nuclear magnetic resonance (NMR) spectroscopy of urine, uses spectral patterns of NMN, 4PY and other endogenous metabolites to predict hepatocellular peroxisome count. Each treatment induced pharmacological (serum lipid) effects characteristic of their class, but only fenofibrate, DEHP and simvastatin increased peroxisome number and raised urinary NMN, 2PY and 4PY, with simvastatin having only a transient effect on the latter. These compounds also reduced mRNA expression for aminocarboxymuconate-semialdehyde decarboxylase (ACMSDase, EC 4.1.1.45), the enzyme believed to be involved in modulating the flux of tryptophan through this pathway, with decreasing order of potency, fenofibrate (-10.39-fold) >DEHP (-3.09-fold) >simvastatin (-1.84-fold). Of the other treatments, only LCFA influenced mRNA expression of ACMSDase (-3.62-fold reduction) and quinolinate phosphoribosyltransferase (QAPRTase, EC 2.4.2.19) (-2.42-fold) without any change in urinary NMN excretion. Although there were no correlations between urinary NMN concentration and serum lipid parameters, NMN did correlate with peroxisome count (r2=0.63) and acyl-CoA oxidase activity (r2=0.61). These correlations were biased by the large response to fenofibrate compared to the other treatments; nevertheless the data do indicate a relationship between the tryptophan-NAD+ pathway and PPARalpha-dependent pathways, making this metabolite a potentially useful biomarker to detect PP. In order to strengthen the observed link between the metabolites associated with the tryptophan-NAD+ pathway and more accurately predict PP, other urinary metabolites were included in a predictive statistical model. This statistical model was found to predict the observed PP in 26/27 instances using a pre-determined threshold of 2-fold mean control peroxisome count. The model also predicted a time-dependent increase in peroxisome count for the fenofibrate group, which is important when considering the use of such modelling to predict the onset and progression of PP prior to its observation in samples taken at autopsy.
authors.affiliation: Safety Assessment, GlaxoSmithKline, Park Road, Ware, Herts, SG12 0DP, UK.
authors.forename: Jane
authors.initials: J
authors.lastname: Delaney
authors.forename: Mark P
authors.initials: MP
authors.lastname: Hodson
authors.forename: Hansa
authors.initials: H
authors.lastname: Thakkar
authors.forename: Susan C
authors.initials: SC
authors.lastname: Connor
authors.forename: Brian C
authors.initials: BC
authors.lastname: Sweatman
authors.forename: Steve P
authors.initials: SP
authors.lastname: Kenny
authors.forename: Paul J
authors.initials: PJ
authors.lastname: McGill
authors.forename: Julie C
authors.initials: JC
authors.lastname: Holder
authors.forename: Kathryn A
authors.initials: KA
authors.lastname: Hutton
authors.forename: John N
authors.initials: JN
authors.lastname: Haselden
authors.forename: Catherine J
authors.initials: CJ
authors.lastname: Waterfield
mesh_terms: D000818:Animals; D015415:Biomarkers; D002262:Carboxy-Lyases / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D015870:Gene Expression; D022781:Hepatocytes / Q000648:ultrastructure; D000960:Hypolipidemic Agents / Q000494:pharmacology; D007150:Immunohistochemistry; D008055:Lipids / Q000097:blood; D008074:Lipoproteins / Q000097:blood; D008111:Liver Function Tests; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D013058:Mass Spectrometry; D046529:Microscopy, Electron, Transmission; D015233:Models, Statistical; D009243:NAD / Q000378:metabolism*; D009929:Organ Size / Q000187:drug effects; D010430:Pentosyltransferases / Q000378:metabolism; D020025:Peroxisome Proliferators / Q000378:metabolism*; D020675:Peroxisomes / Q000648:ultrastructure; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D014364:Tryptophan / Q000378:metabolism*; D000098553:Nicotinate-Nucleotide Diphosphorylase (Carboxylating)
chemical_list: D015415:Biomarkers; D000960:Hypolipidemic Agents; D008055:Lipids; D008074:Lipoproteins; D020025:Peroxisome Proliferators; D012333:RNA, Messenger; D009243:NAD; D014364:Tryptophan; D010430:Pentosyltransferases; D000098553:Nicotinate-Nucleotide Diphosphorylase (Carboxylating); D002262:Carboxy-Lyases; C024396:aminocarboxymuconate-semialdehyde decarboxylase