Urinary Amine Metabolomics Characterization with Custom 12-Plex Isobaric DiLeu Labeling

Area of Science:

• Metabolomics
• Urology
• Biomarker Discovery

Background:

• Lower urinary tract symptoms (LUTS) are prevalent in aging males, with unknown causes and subjective diagnostics.
• Current LUTS treatments are often ineffective and burdensome, highlighting the need for objective criteria.
• Biomarker discovery is crucial for personalized medicine and identifying new therapeutic targets in LUTS.

Purpose of the Study:

• To identify urine metabolite markers specific to hormone-induced bladder outlet obstruction.
• To investigate the temporal changes in the metabolome during the development of LUTS in a mouse model.
• To establish an efficient workflow for amine-containing metabolomics in biological specimens.

Main Methods:

• Utilized a mouse model of LUTS induced by steroid hormone treatment.
• Collected urine samples at baseline and after 2, 4, and 8 weeks of treatment.
• Applied 12-plex DiLeu isobaric labeling and nanoflow ultrahigh-performance liquid chromatography-tandem mass spectrometry for metabolite quantification.
• Employed Metandem software for data processing and metabolite identification.

Main Results:

• Quantified 59 amine-containing metabolites, with 9 significantly altered by hormone treatment.
• Identified dysregulation in three metabolic pathways, notably arginine and proline metabolism.
• Observed significant changes in proline and citrulline in both the mouse model and human LUTS patient samples.

Conclusions:

• Proline and citrulline are promising candidate biomarkers for hormone-induced LUTS.
• The 12-plex DiLeu labeling workflow combined with Metandem is an effective method for amine metabolomics.
• Objective biomarkers could advance personalized medicine and treatment strategies for LUTS.