Discovery of Candidate Stool Biomarker Proteins for Biliary Atresia Using Proteome Analysis by Data-Independent Acquisition Mass Spectrometry.

Eiichiro Watanabe,Akira Nishi,Kan Suzuki,Jun Fujishiro,Ren Nakamura,Shinya Takazawa,Tomo Kakihara,Wataru Suda,Osamu Ohara,Daisuke Nakajima,Yusuke Kawashima

doi:10.3390/proteomes8040036

Abstract

Biliary atresia (BA) is a destructive inflammatory obliterative cholangiopathy of the neonate that affects various parts of the bile duct. If early diagnosis followed by Kasai portoenterostomy is not performed, progressive liver cirrhosis frequently leads to liver transplantation in the early stage of life. Therefore, prompt diagnosis is necessary for the rescue of BA patients. However, the prompt diagnosis of BA remains challenging because specific and reliable biomarkers for BA are currently unavailable. In this study, we discovered potential biomarkers for BA using deep proteome analysis by data-independent acquisition mass spectrometry (DIA–MS). Four patients with BA and three patients with neonatal cholestasis of other etiologies (non-BA) were recruited for stool proteome analysis. Among the 2110 host-derived proteins detected in their stools, 49 proteins were significantly higher in patients with BA and 54 proteins were significantly lower. These varying stool protein levels in infants with BA can provide potential biomarkers for BA. As demonstrated in this study, the deep proteome analysis of stools has great potential not only in detecting new stool biomarkers for BA but also in elucidating the pathophysiology of BA and other pediatric diseases, especially in the field of pediatric gastroenterology.

Highlights

Biliary atresia (BA) is a destructive inflammatory obliterative cholangiopathy of neonates that affects various parts of the intra- and extrahepatic bile duct and causes cholestasis, which manifests as jaundice with hyperbilirubinemia
BA were classified as type III, whereas three non-BA patients had neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD), cholestasis after repair of gastroschisis, and veno-occlusive disease (VOD)
We found that specific proteins that are elevated in the liver tissues, such as RBP4, SHMT2, HMGCS1, ADH6, ALDH1A1, ACADS, adenosine kinase (ADK), KHK, ACAA2, PSAT1, AMACR, and prostaglandin reductase 1 (PTGR1), presented significantly lower abundances in stool samples from patients with BA

Summary

Introduction

Biliary atresia (BA) is a destructive inflammatory obliterative cholangiopathy of neonates that affects various parts of the intra- and extrahepatic bile duct and causes cholestasis, which manifests as jaundice with hyperbilirubinemia. Stool is a feasible clinical sample for exploring biomarkers of gastrointestinal diseases such as BA. This expectation originated from one of the clinical symptoms of cholestasis, such as BA, in which the stool color gradually changed from normal (yellowish, brown, and greenish) to abnormal (clay-colored, pale yellowish, and light yellowish), followed by a gradual reduction in bile supply from the liver and gallbladder [7]. Host proteins in the stool of patients with colorectal cancer have been examined using proteomics, leading to the discovery of candidate colon biomarkers [8,9,10]. High-depth proteome analysis is required to detect more host-derived proteins in stools

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Results

Conclusion