title: Noninvasive urinary metabolomic profiling identifies diagnostic and prognostic markers in lung cancer.
abstract: Lung cancer remains the most common cause of cancer deaths worldwide, yet there is currently a lack of diagnostic noninvasive biomarkers that could guide treatment decisions. Small molecules (<1,500 Da) were measured in urine collected from 469 patients with lung cancer and 536 population controls using unbiased liquid chromatography/mass spectrometry. Clinical putative diagnostic and prognostic biomarkers were validated by quantitation and normalized to creatinine levels at two different time points and further confirmed in an independent sample set, which comprises 80 cases and 78 population controls, with similar demographic and clinical characteristics when compared with the training set. Creatine riboside (IUPAC name: 2-{2-[(2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)-oxolan-2-yl]-1-methylcarbamimidamido}acetic acid), a novel molecule identified in this study, and N-acetylneuraminic acid (NANA) were each significantly (P < 0.00001) elevated in non-small cell lung cancer and associated with worse prognosis [HR = 1.81 (P = 0.0002), and 1.54 (P = 0.025), respectively]. Creatine riboside was the strongest classifier of lung cancer status in all and stage I-II cases, important for early detection, and also associated with worse prognosis in stage I-II lung cancer (HR = 1.71, P = 0.048). All measurements were highly reproducible with intraclass correlation coefficients ranging from 0.82 to 0.99. Both metabolites were significantly (P < 0.03) enriched in tumor tissue compared with adjacent nontumor tissue (N = 48), thus revealing their direct association with tumor metabolism. Creatine riboside and NANA may be robust urinary clinical metabolomic markers that are elevated in tumor tissue and associated with early lung cancer diagnosis and worse prognosis.
authors.affiliation: Authors' Affiliations: Laboratory of Molecular Immunogenomics, Genomic and Immunity Section, NIAMS/NIH; Laboratories of Human Carcinogenesis, and Metabolism, National Cancer Institute, NIH, Bethesda, Maryland; Department of Veterinary and Biomedical Sciences and Center for Molecular Toxicology and Carcinogenesis; Metabolomics Core Facility; Nuclear Magnetic Resonance Spectroscopy, The Pennsylvania State University, University Park, Pennsylvania; Ohio State University Comprehensive Cancer Center, Columbus, Ohio; and Department of Clinical Research, University of Bern, Bern, SwitzerlandAuthors' Affiliations: Laboratory of Molecular Immunogenomics, Genomic and Immunity Section, NIAMS/NIH; Laboratories of Human Carcinogenesis, and Metabolism, National Cancer Institute, NIH, Bethesda, Maryland; Department of Veterinary and Biomedical Sciences and Center for Molecular Toxicology and Carcinogenesis; Metabolomics Core Facility; Nuclear Magnetic Resonance Spectroscopy, The Pennsylvania State University, University Park, Pennsylvania; Ohio State University Comprehensive Cancer Center, Columbus, Ohio; and Department of Clinical Research, University of Bern, Bern, Switzerland.
authors.forename: Ewy A
authors.initials: EA
authors.lastname: Mathé
authors.affiliation: Authors' Affiliations: Laboratory of Molecular Immunogenomics, Genomic and Immunity Section, NIAMS/NIH; Laboratories of Human Carcinogenesis, and Metabolism, National Cancer Institute, NIH, Bethesda, Maryland; Department of Veterinary and Biomedical Sciences and Center for Molecular Toxicology and Carcinogenesis; Metabolomics Core Facility; Nuclear Magnetic Resonance Spectroscopy, The Pennsylvania State University, University Park, Pennsylvania; Ohio State University Comprehensive Cancer Center, Columbus, Ohio; and Department of Clinical Research, University of Bern, Bern, Switzerland.
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Patterson
authors.affiliation: Authors' Affiliations: Laboratory of Molecular Immunogenomics, Genomic and Immunity Section, NIAMS/NIH; Laboratories of Human Carcinogenesis, and Metabolism, National Cancer Institute, NIH, Bethesda, Maryland; Department of Veterinary and Biomedical Sciences and Center for Molecular Toxicology and Carcinogenesis; Metabolomics Core Facility; Nuclear Magnetic Resonance Spectroscopy, The Pennsylvania State University, University Park, Pennsylvania; Ohio State University Comprehensive Cancer Center, Columbus, Ohio; and Department of Clinical Research, University of Bern, Bern, Switzerland.
authors.forename: Majda
authors.initials: M
authors.lastname: Haznadar
authors.affiliation: Authors' Affiliations: Laboratory of Molecular Immunogenomics, Genomic and Immunity Section, NIAMS/NIH; Laboratories of Human Carcinogenesis, and Metabolism, National Cancer Institute, NIH, Bethesda, Maryland; Department of Veterinary and Biomedical Sciences and Center for Molecular Toxicology and Carcinogenesis; Metabolomics Core Facility; Nuclear Magnetic Resonance Spectroscopy, The Pennsylvania State University, University Park, Pennsylvania; Ohio State University Comprehensive Cancer Center, Columbus, Ohio; and Department of Clinical Research, University of Bern, Bern, Switzerland.
authors.forename: Soumen K
authors.initials: SK
authors.lastname: Manna
authors.affiliation: Authors' Affiliations: Laboratory of Molecular Immunogenomics, Genomic and Immunity Section, NIAMS/NIH; Laboratories of Human Carcinogenesis, and Metabolism, National Cancer Institute, NIH, Bethesda, Maryland; Department of Veterinary and Biomedical Sciences and Center for Molecular Toxicology and Carcinogenesis; Metabolomics Core Facility; Nuclear Magnetic Resonance Spectroscopy, The Pennsylvania State University, University Park, Pennsylvania; Ohio State University Comprehensive Cancer Center, Columbus, Ohio; and Department of Clinical Research, University of Bern, Bern, Switzerland.
authors.forename: Kristopher W
authors.initials: KW
authors.lastname: Krausz
authors.affiliation: Authors' Affiliations: Laboratory of Molecular Immunogenomics, Genomic and Immunity Section, NIAMS/NIH; Laboratories of Human Carcinogenesis, and Metabolism, National Cancer Institute, NIH, Bethesda, Maryland; Department of Veterinary and Biomedical Sciences and Center for Molecular Toxicology and Carcinogenesis; Metabolomics Core Facility; Nuclear Magnetic Resonance Spectroscopy, The Pennsylvania State University, University Park, Pennsylvania; Ohio State University Comprehensive Cancer Center, Columbus, Ohio; and Department of Clinical Research, University of Bern, Bern, Switzerland.
authors.forename: Elise D
authors.initials: ED
authors.lastname: Bowman
authors.affiliation: Authors' Affiliations: Laboratory of Molecular Immunogenomics, Genomic and Immunity Section, NIAMS/NIH; Laboratories of Human Carcinogenesis, and Metabolism, National Cancer Institute, NIH, Bethesda, Maryland; Department of Veterinary and Biomedical Sciences and Center for Molecular Toxicology and Carcinogenesis; Metabolomics Core Facility; Nuclear Magnetic Resonance Spectroscopy, The Pennsylvania State University, University Park, Pennsylvania; Ohio State University Comprehensive Cancer Center, Columbus, Ohio; and Department of Clinical Research, University of Bern, Bern, Switzerland.
authors.forename: Peter G
authors.initials: PG
authors.lastname: Shields
authors.affiliation: Authors' Affiliations: Laboratory of Molecular Immunogenomics, Genomic and Immunity Section, NIAMS/NIH; Laboratories of Human Carcinogenesis, and Metabolism, National Cancer Institute, NIH, Bethesda, Maryland; Department of Veterinary and Biomedical Sciences and Center for Molecular Toxicology and Carcinogenesis; Metabolomics Core Facility; Nuclear Magnetic Resonance Spectroscopy, The Pennsylvania State University, University Park, Pennsylvania; Ohio State University Comprehensive Cancer Center, Columbus, Ohio; and Department of Clinical Research, University of Bern, Bern, Switzerland.
authors.forename: Jeffrey R
authors.initials: JR
authors.lastname: Idle
authors.affiliation: Authors' Affiliations: Laboratory of Molecular Immunogenomics, Genomic and Immunity Section, NIAMS/NIH; Laboratories of Human Carcinogenesis, and Metabolism, National Cancer Institute, NIH, Bethesda, Maryland; Department of Veterinary and Biomedical Sciences and Center for Molecular Toxicology and Carcinogenesis; Metabolomics Core Facility; Nuclear Magnetic Resonance Spectroscopy, The Pennsylvania State University, University Park, Pennsylvania; Ohio State University Comprehensive Cancer Center, Columbus, Ohio; and Department of Clinical Research, University of Bern, Bern, Switzerland.
authors.forename: Philip B
authors.initials: PB
authors.lastname: Smith
authors.affiliation: Authors' Affiliations: Laboratory of Molecular Immunogenomics, Genomic and Immunity Section, NIAMS/NIH; Laboratories of Human Carcinogenesis, and Metabolism, National Cancer Institute, NIH, Bethesda, Maryland; Department of Veterinary and Biomedical Sciences and Center for Molecular Toxicology and Carcinogenesis; Metabolomics Core Facility; Nuclear Magnetic Resonance Spectroscopy, The Pennsylvania State University, University Park, Pennsylvania; Ohio State University Comprehensive Cancer Center, Columbus, Ohio; and Department of Clinical Research, University of Bern, Bern, Switzerland.
authors.forename: Katsuhiro
authors.initials: K
authors.lastname: Anami
authors.affiliation: Authors' Affiliations: Laboratory of Molecular Immunogenomics, Genomic and Immunity Section, NIAMS/NIH; Laboratories of Human Carcinogenesis, and Metabolism, National Cancer Institute, NIH, Bethesda, Maryland; Department of Veterinary and Biomedical Sciences and Center for Molecular Toxicology and Carcinogenesis; Metabolomics Core Facility; Nuclear Magnetic Resonance Spectroscopy, The Pennsylvania State University, University Park, Pennsylvania; Ohio State University Comprehensive Cancer Center, Columbus, Ohio; and Department of Clinical Research, University of Bern, Bern, Switzerland.
authors.forename: Dickran G
authors.initials: DG
authors.lastname: Kazandjian
authors.affiliation: Authors' Affiliations: Laboratory of Molecular Immunogenomics, Genomic and Immunity Section, NIAMS/NIH; Laboratories of Human Carcinogenesis, and Metabolism, National Cancer Institute, NIH, Bethesda, Maryland; Department of Veterinary and Biomedical Sciences and Center for Molecular Toxicology and Carcinogenesis; Metabolomics Core Facility; Nuclear Magnetic Resonance Spectroscopy, The Pennsylvania State University, University Park, Pennsylvania; Ohio State University Comprehensive Cancer Center, Columbus, Ohio; and Department of Clinical Research, University of Bern, Bern, Switzerland.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Hatzakis
authors.affiliation: Authors' Affiliations: Laboratory of Molecular Immunogenomics, Genomic and Immunity Section, NIAMS/NIH; Laboratories of Human Carcinogenesis, and Metabolism, National Cancer Institute, NIH, Bethesda, Maryland; Department of Veterinary and Biomedical Sciences and Center for Molecular Toxicology and Carcinogenesis; Metabolomics Core Facility; Nuclear Magnetic Resonance Spectroscopy, The Pennsylvania State University, University Park, Pennsylvania; Ohio State University Comprehensive Cancer Center, Columbus, Ohio; and Department of Clinical Research, University of Bern, Bern, Switzerland.
authors.forename: Frank J
authors.initials: FJ
authors.lastname: Gonzalez
authors.affiliation: Authors' Affiliations: Laboratory of Molecular Immunogenomics, Genomic and Immunity Section, NIAMS/NIH; Laboratories of Human Carcinogenesis, and Metabolism, National Cancer Institute, NIH, Bethesda, Maryland; Department of Veterinary and Biomedical Sciences and Center for Molecular Toxicology and Carcinogenesis; Metabolomics Core Facility; Nuclear Magnetic Resonance Spectroscopy, The Pennsylvania State University, University Park, Pennsylvania; Ohio State University Comprehensive Cancer Center, Columbus, Ohio; and Department of Clinical Research, University of Bern, Bern, Switzerland Curtis_Harris@nih.gov.
authors.forename: Curtis C
authors.initials: CC
authors.lastname: Harris
mesh_terms: D000368:Aged; D014408:Biomarkers, Tumor / Q000652:urine*; D002289:Carcinoma, Non-Small-Cell Lung / Q000175:diagnosis / Q000401:mortality / Q000652:urine*; D016022:Case-Control Studies; D003401:Creatine / Q000031:analogs & derivatives* / Q000652:urine; D005260:Female; D006801:Humans; D053208:Kaplan-Meier Estimate; D008175:Lung Neoplasms / Q000175:diagnosis / Q000401:mortality / Q000652:urine*; D008297:Male; D055442:Metabolome; D008875:Middle Aged; D019158:N-Acetylneuraminic Acid / Q000652:urine*; D011379:Prognosis; D016016:Proportional Hazards Models; D012372:ROC Curve; D012263:Ribonucleosides / Q000652:urine*; D012907:Smoking / Q000652:urine
chemical_list: C000590396:2-(2-(3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl)-1-methylcarbamimidamido)acetic acid; D014408:Biomarkers, Tumor; D012263:Ribonucleosides; D019158:N-Acetylneuraminic Acid; D003401:Creatine
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