Disease monitoring of hepatocellular carcinoma through metabolomics

Table 1 Significantly altered metabolites in hepatocellular carcinoma patients vs cirrhosis controls

Ref.	Platform	Tissue (organism)	HCC etiology	Significantly altered metabolites in HCC patients vs cirrhosis controls	Main pathways distinguishing HCC from cirrhosis
Patterson et al[29]	UPLC/QTOF-MS; UPLC/ESI-TQMS; GC/MS	Plasma (human)	HCC n = 20:	LPC (14:0) ↓	LPC metabolism
			EtOH n = 6	LPC (18:1) ↓
			HBV n = 3	LPC (20:4) ↓
			HCV n = 5	LPC (20:3) ↓
			NASH n = 3	LPC (22:6) ↓
			NASH/alcoholic steatohepatitis n = 1	FFA (24:0) - lignoceric acid ↓	Very long chain fatty acid metabolism
			HH n = 2	FFA (24:1) - nervonic acid ↓	Hemoglobin metabolism
				Bilirubin ↑
				Biliverdin ↑
Xiao et al[37]	UPLC/QTOF-MS	Serum (human)	HCV	3β, 6β-dihydroxy-5β-cholan-24-oic acid ↓	Bile acid biosynthesis
				3α, 7β-dihydroxy-5β-cholest-24-en-26-oic acid ↓
				GCA ↓
				GDCA ↓
				GCDCA ↓
				TCDCA ↓
				Linoelaidyl carnitine ↓	CPT shuttle system
				Oleoylcarnitine ↓
				Palmitoyl carnitine ↓
				O-octanoyl-R-carnitine ↓
				LPC (20:1) ↓	LPC metabolism
				LPC (20:4) ↓
				PE (20:4/18:1) ↓
				4E;15Z-Bilirubin IXa ↓	Hemoglobin metabolism
				15,16-dihydrobiliverdin ↓
				3-ganidinopropionicacid ↓
				Tetracosahexaenoic acid ↓	Eicosanoid metabolism
				3-hydroxy-eicosanoic acid ↓
				Oleamide ↓
				Phe-Phe ↑
Wang et al[38]	UPLC/MS-MS; LC/QTOF-MS	Serum (human)	HCC n = 82:	LPC-16:0 ↑	LPC metabolism
			HBV-cirrhosis n = 41	LPC-18:0 ↑
			HBV-only n = 41	16:0/18:1-PC ↑
				16:0/18:2-PC ↑
				16:0/20:4-PC ↑
				16:0/22:6-PC ↑
				18:0/18:2-PC ↑
				Phenylalanine ↓	Gut flora metabolism
				GCDCA ↓	Bile acid metabolism
				Canavaninosuccinate ↑↑	Organic acid metabolism
Zhou et al[39]	UPLC/QTOF-MS	Serum (human)	HCC n = 69:	LPC (14:0) HCC-C ↓	LPC metabolism
			HBV-HCC (HCC-B) n = 38	LPC (16:1) HCC-C ↓
			HCV-HCC (HCC-C) n = 31	LPC (18:3) HCC-B ↓
				HCC-C ↓
				LPC (18:2) HCC-B ↓
				HCC-C ↓
				LPC (18:1) HCC-B ↓
				HCC-C ↓
				LPC (18:0) HCC-C ↓
				LPC (20:5) HCC-C ↓
				LPC (20:4) HCC-B ↓
				HCC-C ↓
				LPC (20:3) HCC-B ↓
				HCC-C ↓
				LPC (20:2) HCC-C ↓
				LPC (20:0) HCC-B ↓
				HCC-C ↓
				LPC (22:6) HCC-C ↓
				LPC (22:5) HCC-B ↓
				HCC-C ↓
				LPC (22:4) HCC-B ↓
				HCC-C ↓
Wu et al[31]	SELDI-TOF-MS; HPLC/MS	Serum (human)	HBV	GRO-α↑	Cytokine
				Thrombin light chain ↑	Protease cleavage
Ressom et al[36]	UPLC/QTOF-MS	Serum (human)	HCC n = 78:	GDCA ↓	Bile acid metabolism
			→ HCV 67%	TCA ↓
			→ HBV 15%	TCDCA ↓
			→ Alcoholism 29%	Sphingosine 1-phosphate ↑	Sphingolipid metabolism
			→ NASH 13%	LPC (16:0) ↑	LPC metabolism
			→ Cryptogenic (8%)	LPC (17:0) ↑
			→ Autoimmune (3%)	LPC (18:0) ↑
				LPC (15:0) ↑
				LPC (22:6) ↑
				LPE (22:6) ↑
				LPE (20:4) ↑
				LPE (20:3) ↑
				PS ↑
Yang et al[28]	HRMAS 1H NMR	Biopsy (human)	HCC n = 17:	Glucose ↓	Glycolysis
			→ Cirrhosis n = 9	Creatine ↓
			→ No cirrhosis n = 8	PE ↑	LPC metabolism
				Glutamine ↑	Amino acid metabolism
				Glutamate ↑
				PC + GPC ↑	Bile acid metabolism
Nahon et al[40]	NMR	Serum (human)	EtOH cirrhosis	High density lipoproteins	HDL biosynthesis
				Acetate ↑	Ketone body metabolism
				N-acetyl-glycoproteins ↑	N-acetylglycoprotein
				Glutamate ↑	Amino acid metabolism
				Glutamine ↓
Budhu et al[43]	GC/MS, UPLC/MS-MS	Biopsy samples (human)	HCC n = 356	Study reported on markers involved in cancer aggressivity through comparison of stem-like HCC to less benign mature hepatocyte HCC	N/A
			Training cohort n = 30
			Testing cohort n = 217
			Validation cohort n = 139
Beyoğlu et al[44]	GC/MS	Biopsy samples (human)	Six HCC subtypes, liver fibrosis status unknown	Glucose ↓	Glycolysis
				Glycerol 3-phosphate ↓
				Glycerol 2-phosphate ↓
				Malate ↓
				Alanine ↓
				Myo-inositol ↓	PI3K pathway
				Linoleic acid ↓	Prostaglandin biosynthesis
Fitian et al[45]	UPLC/MS-MS and GC/MS	Serum (human)	HCV cirrhosis-associated HCC n = 30	Sphingosine ↑	Sphingolipid
			HCV-cirrhosis n = 27	Xanthine ↑	Oxidative stress metabolism
			Healthy volunteers n = 30	2-Pyrrolidinone ↑	GABA metabolism
				2-Hydroxybutyrate ↑	Oxidative stress metabolism
				Serine ↑	Amino acid
				Glycine ↑
				Aspartate ↑
				12-HETE ↑	Inflammation pathway
				15-HETE ↑
				Isovalerate ↑	Gut microflora metabolism
				Dihomo-linolenate ↑	Inflammation pathway
Gao et al[46]	GC-TOF/MS	Serum (human)	HBV cirrhosis-associated HCC n = 39	Stearic acid	Fatty acid biosynthesis
			HBV-cirrhosis (n = 52)	Heptadecanoic acid
				Palmitic acid
				5-Aminovaleric acid	Gut microflora metabolism
				Cholesterol ↑	Cholesterol metabolism
				3-hydroxybutyric acid ↑	Ketogenesis
				Malic acid ↑	TCA metabolism
				Glutamine ↑	Amino acid
				Asparagine ↓
				Alanine ↑
				Threonine ↓
				Leucine ↓
				Glutamic acid ↑
				β-glutamate ↑
				5-oxoproline ↓	Glutathione metabolism
				1,2,4-cyclopropranodicarboxylic acid ↓	Dicarboxylic acid metabolism

Pathways of importance in the comparison of (1A) HCC vs cirrhosis and (2) cirrhosis vs healthy controls are shown. Arrows indicate the metabolite’s expression in cases vs appropriate controls. P < 0.05 was used as the significance level and metabolites reported in table are those which were most significantly upregulated or downregulated in each study. EtOH: Alcohol; TOCSY: Total correlation spectroscopy; HH: Hereditary hemochromatosis; TCA: Tricarboxylic acid; UPLC: Ultrahigh-performance liquid chromatography; QTOF: Quadrupole time of flight; SELDI: Surface-enhanced laser desorption/ionization; HRMAS: High-resolution magic angle spinning; LPE: Lysophosphatidyethanolamine; LPC: Lysophosphatidylcholine; HCC: Hepatocellular carcinoma; MS: Mass spectrometry; TOF: Time-of-flight; GC: Gas chromatography; LC: Liquid chromatography; NMR: Nuclear magnetic resonance; HBV: Hepatitis B virus; HCV: Hepatitis C virus; NASH: Non-alcoholic steatohepatitis; FFA: Free fatty acids; PE: Phosphorylethanolamine; GABA: γ-aminobutyric acid.