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Analysis and Comparison of the composition and metabolic function of the intestinal microbiome of tumor patients with and without tumor cachexia: a prospective, non-interventional, explorative cohort-study in patients with gastrointestinal tumor stage UICC III-IV

ABSTRACT: Interventions: Group 1: Arm 1: Blood-, urine- and stool-samples of 60 patients with gastrointestinal tumors (UICC III-IV) without tumor cachexia will be frozen. The samples will be collected and send to the analysis lab for analysis. In addition, a questionnaire will be completed by the patient. Group 2: Arm 2: Blood-, urine- and stool-samples of 60 patients with gastrointestinal tumors (UICC III-IV) with tumor cachexia will be frozen. The samples will be collected and send to the analysis lab for analysis. In addition, a questionnaire will be completed by the patient. Primary outcome(s): - Evaluation of intestinal genes in stool samples of patients using next-generation sequencing technology - Identification of differences regarding the diversity of intestinal microorganisms between both groups Primary and endpoint will be assessed once based on the collected samples. No follow-up samples will be collected. Study Design: Allocation: ; Masking: ; Control: ; Assignment: ; Study design purpose: basic science

DISEASE(S): Malignant Neoplasm, Primary Site Unspecified,Cachexia

PROVIDER: 2433991 | ecrin-mdr-crc |

REPOSITORIES: ECRIN MDR

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Project description:BACKGROUND: Stool metabolites provide essential insights into the function of the gut microbiome. The current gold standard for storage of stool samples for metabolomics is flash-freezing at - 80 °C which can be inconvenient and expensive. Ambient temperature storage of stool is more practical, however no available methodologies adequately preserve the metabolomic profile of stool. A novel sampling kit (OMNImet.GUT; DNA Genotek, Inc.) was introduced for ambient temperature storage and stabilization of feces for metabolomics; we aimed to test the performance of this kit vs flash-freezing. To do this stool was collected from an infant's diaper was divided into 2 aliquots: 1) flash-frozen and 2) stored in an OMNImet.GUT tube at ambient temperature for 3-4 days. Samples from the same infant were collected at 2 different timepoints to assess metabolite changes over time. Subsequently, all samples underwent metabolomic analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS).RESULTS: Paired fecal samples (flash-frozen and ambient temperature) from 16 infants were collected at 2 timepoints (32 individual samples, 64 aliquots). Similar numbers of metabolites were detected in both the frozen and ambient temperature samples (1126 in frozen, 1107 in ambient temperature, 1064 shared between sample types). Metabolite abundances were strongly correlated between storage methods (median Spearman correlation Rs = 0.785 across metabolites). Hierarchical clustering analysis and principal component analysis showed that samples from the same individuals at a given timepoint clustered closely, regardless of the storage method. Repeat samples from the same individual were compared by paired t-test, separately for the frozen and OMNImet.GUT. The number of metabolites in each biochemical class that significantly changed (p < 0.05) at timepoint 2 relative to timepoint 1 was similar in flash-frozen vs ambient temperature storage. Changes in microbiota modified metabolites over time were also consistent across both methodologies.CONCLUSION: Ambient temperature storage and stabilization of stool in the OMNImet.GUT device yielded comparable metabolomic results to flash freezing in terms of 1) the identity and abundance of detected biochemicals 2) the distinct metabolomic profiles of subjects and 3) changes in metabolites over time that are plausibly microbiota-induced. This method potentially provides a more convenient, less expensive home collection and storage option for stool metabolomic analysis.

Dataset Information

Analysis and Comparison of the composition and metabolic function of the intestinal microbiome of tumor patients with and without tumor cachexia: a prospective, non-interventional, explorative cohort-study in patients with gastrointestinal tumor stage UICC III-IV

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