Metabolomics

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Metabolic Footprinting of a Clear Cell Renal Cell Carcinoma in vitro Model for Human Kidney Cancer Detection


ABSTRACT: A protocol for harvesting and extracting extracellular metabolites from an in vitro model of human renal cell lines was developed to profile the exometabolome by means of a discovery-based metabolomics approach using ultraperformance liquid chromatography coupled to quadrupole-time-of-flight mass spectrometry. Metabolic footprints provided by conditioned media (CM) samples (n=66) of two clear cell Renal Cell Carcinoma (ccRCC) cell lines with different genetic background and a non-tumor renal cell line, were compared with the human serum metabolic profile of a pilot cohort (n=10) comprised of stage IV ccRCC patients and healthy individuals. Using a cross-validated orthogonal projection to latent structures-discriminant analysis model, a panel of 21 discriminant features selected by iterative multivariate classification, allowed differentiating control from tumor cell lines with 100% specificity, sensitivity and accuracy. Isoleucine/leucine, phenylalanine, N-lactoyl-leucine, and N-acetyl-phenylalanine, and cysteinegluthatione disulfide (CYSSG) were identified by chemical standards, and hydroxyprolyl-valine was identified with MS and MS/MS experiments. A subset of 9 discriminant features, including the identified metabolites except for CYSSG, produced a fingerprint of classification value that enabled discerning ccRCC patients from healthy individuals. To our knowledge, this is the first time that N-lactoyl-leucine is associated to ccRCC. Results from this study provide a proof of concept that CM can be used as a serum proxy to obtain disease-related metabolic signatures.

INSTRUMENT(S): Xevo G2S QTOF mass spectrometer (Waters), Xevo G2S QTOF mass spectrometer (Waters Corp.)

SUBMITTER: Malena Manzi 

PROVIDER: MTBLS737 | MetaboLights | 2018-11-14

REPOSITORIES: MetaboLights

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Metabolic Footprinting of a Clear Cell Renal Cell Carcinoma in Vitro Model for Human Kidney Cancer Detection.

Knott María Elena ME   Manzi Malena M   Zabalegui Nicolás N   Salazar Mario O MO   Puricelli Lydia I LI   Monge María Eugenia ME  

Journal of proteome research 20181012 11


A protocol for harvesting and extracting extracellular metabolites from an in vitro model of human renal cell lines was developed to profile the exometabolome by means of a discovery-based metabolomics approach using ultraperformance liquid chromatography coupled to quadrupole-time-of-flight mass spectrometry. Metabolic footprints provided by conditioned media (CM) samples ( n = 66) of two clear cell Renal Cell Carcinoma (ccRCC) cell lines with different genetic backgrounds and a nontumor renal  ...[more]

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