Tumour metabolomics and transcriptomics co-expression network as related to biological folate alteration and cancer malignancy remains unexplored in human non-small cell lung cancers (NSCLC). To probe the diagnostic biomarkers, tumour and pair lung tissue samples (n = 56) from 97 NSCLC patients were profiled for ultra-performance liquid chromatography tandem mass spectrometry (UPLC/MS/MS)-analysed metabolomics, targeted transcriptionomics, and clinical folate traits. Weighted Gene Co-expression Network Analysis (WGCNA) was performed. Tumour lactate was identified as the top VIP marker to predict advance NSCLC (AUC = 0.765, Sig = 0.017, CI 0.58–0.95). Low folate (LF)-tumours vs. adjacent lungs displayed higher glycolytic index of lactate and glutamine-associated amino acids in enriched biological pathways of amino sugar and glutathione metabolism specific to advance NSCLCs. WGCNA classified the green module for hub serine-navigated glutamine metabolites inversely associated with tumour and RBC folate, which module metabolites co-expressed with a predominant up-regulation of LF-responsive metabolic genes in glucose transport (GLUT1), de no serine synthesis (PHGDH, PSPH, and PSAT1), folate cycle (SHMT1/2 and PCFR), and down-regulation in glutaminolysis (SLC1A5, SLC7A5, GLS, and GLUD1). The LF-responsive WGCNA markers predicted poor survival rates in lung cancer patients, which could aid in optimizing folate intervention for better prognosis of NSCLCs susceptible to folate malnutrition.

Objectives Lung cancer is the leading cause of cancer-related mortality in the worldwide. In non-small-cell lung cancers (NSCLC), metabolic reprogramming of lactate metabolism has been proposed as a key player in cancer metastasis. Nutritional status of methyl donors was associated with increased risks of lung cancers, yet their roles in lactate metabolism and metastatic NSCLC development remains unclear. Methods The cross-sectional study recruited 100 NSCLC patients with selected 18 paired NSCLC tissues from National Taiwan University Hospital (NTUH), Taiwan. Plasma methyl donors levels (folate/free choline/betaine) of NSCLC patients and target metabolomics signatures of paired NSCLC tissues were analyzed by Liquid chromatography–mass spectrometry (LC/MS). Clinical data of NSCLC tissues were collected from the department of pathology in NTUH. Results Partial Least Squares Discriminant Analysis (PLSDA) revealed that metabolic signatures of tumor/non-tumor lung tissues were well segregated. The two major tumor metabolites signatures of metastatic NSCLC were lactate and glucose, the levels of which were high and low, respectively, with both metabolites displaying the two highest VIP scores. A correlation heatmap showed that tumor lactate levels were strongly associated with increased TCA metabolites (pyruvate, succinate, fumarate and malate) and anaplerotic amino acid levels (alanine and arginine), and inversely correlated with glycolytic metabolites (glucose and PEP). When stratifying plasma methyl donors status of 18 paired NSCLC tumors with metabolomics VIP scores, threshold levels of folate (≥6 ng/ml), free choline (≥9.78 µmol/L) and betaine (≥62.0 µmol/L) were associated with high lactate and low glucose signatures of NSCLC. In particular, plasma betaine was positively associated with tumor lactate (r = 0.51, P = 0.031); plasma lactate was positively associated with tumor glucose (r = 0.53, P = 0.023). Conclusions Our data demonstrate that metastatic NSCLCs were signified with high-lactate metabolizing-metabolomics fingerprints which were modified by plasma methyl donors’ status with a differential threshold effect. Funding Sources This study was supported by the three-year grant from the Ministration of Science and Technology, Taiwan, ROC.

In humans, PEMT rs7946 polymorphism exerts sex-specific effects on choline requirement and hepatic steatosis (HS) risk. Few studies have explored the interaction effect of the PEMT rs7946 polymorphism and sex on the effect of adequate choline intake on HS risk. In this cross-sectional study, we investigated the association between PEMT polymorphism and adequate choline intake on HS risk. We enrolled 250 older patients with metabolic disorders with (n = 152) or without (n = 98; control) ultrasonically diagnosed HS. An elevated PEMT rs7946 A allele level was associated with a lower HS risk and body mass index in both men and women. Dietary choline intake—assessed using a semiquantitative food frequency questionnaire—was associated with reduced obesity in men only (p for trend < 0.05). ROC curve analysis revealed that the cutoff value of energy-adjusted choline intake for HS diagnosis was 448 mg/day in women (AUC: 0.62; 95% CI: 0.57–0.77) and 424 mg/day in men (AUC: 0.63, 95% CI: 0.57–0.76). In women, GG genotype and high choline intake (>448 mg/day) were associated with a 79% reduction in HS risk (adjusted OR: 0.21; 95% CI: 0.05–0.82); notably, GA or AA genotype was associated with a reduced HS risk regardless of choline intake (p < 0.05). In men, GG genotype and high choline intake (>424 mg/day) were associated with a 3.7-fold increase in HS risk (OR: 3.7; 95% CI: 1.19–11.9). Further adjustments for a high-density lipoprotein level and body mass index mitigated the effect of choline intake on HS risk. Current dietary choline intake may be inadequate for minimizing HS risk in postmenopausal Taiwanese women carrying the PEMT rs7946 GG genotype. Older men consuming more than the recommended amount of choline may have an increased risk of nonalcoholic fatty liver disease; this risk is mediated by a high-density lipoprotein level and obesity.