Proof-of-Concept Workflow for Establishing Reference Intervals of Human Urine Proteome for Monitoring Physiological and Pathological Changes

Abstract

Urine as a true non-invasive sampling source holds great potential for biomarker discovery. While approximately 2000 proteins can be detected by mass spectrometry in urine from healthy people, the amount of these proteins vary considerably. A systematic evaluation of a large number of samples is needed to determine the range of the variations. Current biomarker studies often measure limited number of urine samples in the discovery phase, which makes it difficult to determine whether proteins differentially expressed between control and disease groups represent actual difference, or are just physiological variations among the individuals, leads to failures in the validation phase with the increased sample numbers. Here, we report a streamlined workflow with capacity of measuring 8 urine proteomes per day at the coverage of > 1500 proteins. With this workflow, we evaluated variations in 497 urine proteomes from 167 healthy donors, establishing reference intervals (RIs) that covered urine protein variations. We demonstrated that RIs could be used to monitor physiological changes by detecting transient outlier proteins. Furthermore, we provided a RIs-based algorithm for biomarker discovery and validation to screen for diseases such as cancer. This study provided a proof-of-principle workflow for the use of urine proteome for health monitoring and disease screening.

Investigation of the urinary metabolic variations and the application in bladder cancer biomarker discovery

“…24 25 In our work, we evaluated the inter-individual CVs of urine metabolites in 203 normal subjects and 33 BCa patients. 24 25 In our work, we evaluated the inter-individual CVs of urine metabolites in 203 normal subjects and 33 BCa patients.…”

“…24 Recently, Leng et al explored the individual variations of 2000 commonly detected urine proteins in 497 proteome samples, which ranged from 1.01 to 1.19. 25 In our work, we evaluated the inter-individual CVs of urine metabolites in 203 normal subjects and 33 BCa patients. The median CV for 203 normal urine metabolome was 0.941 and for the 33 BCa urine metabolomes, it was 0.833.…”

Urinary proteomics of Henoch-Schönlein purpura nephritis in children using liquid chromatography-tandem mass spectrometry

“…In recent years, urine has been considered a potential source to provide important clues regarding the progression of diseases [6]. The normal excretion of proteins in urine is less than 150 mg/L per 24 h, and approximately 30% of urinary protein originates from plasma via blood filtration, whereas the remaining 70% is derived from the kidneys.…”

Urine Proteome Changes in a TNBS‐Induced Colitis Rat Model

“…However, there have been limited urinary biomarker studies for intestinal diseases, specifically for IBD, compared with the wide applications in renal diseases. Studying biomarkers in clinical samples remains challenging . Studying the urine proteome is especially challenging because it is influenced by multiple factors, such as age, diet, exercise, gender, medication, and daily rhythms .…”