Proteomic Analysis Provides a New Sight Into the CRABP1 Expression in the Pathogenesis of Hirschsprung Disease

Abstract

Hirschsprung’s disease (HSCR) is the most common developmental disorder of the enteric nervous system and its etiology and pathogenesis remain largely unknown. This study aims to identify the differential proteomic patterns linked to the occurrence and development of Hirschsprung disease in colonic tissues. Biopsies were obtained from the aganglionic colon in human HSCR and the corresponding ganglionic colon segments for direct quantitative determination of the data-independent acquisition (DIA) followed by bioinformatics analysis. The differentially expressed main proteins were confirmed by Western blot and immunostaining. A total of 5832 proteins were identified in human colon tissues. Among them, 97 differentially expressed proteins (DEP) with fold change (FC) > 1.2 were screened, including 18 upregulated proteins and 79 downregulated proteins, and GO and KEGG enrichment analyses were performed on differential proteins. By comparing down-regulated proteins with highly connected protein nodes in the PPI network with those related to intracellular metabolic processes in the above analysis, we identified cellular retinoic acid binding protein 1(CRABP1). Its expression was verified in the aganglionic part of the colon by western blotting in an expanded sample set (P = 0.0031). The immunostaining results revealed that CRABP1 was highly expressed in the myenteric plexus ganglion in ganglionic colons compared to aganglionic segments (P = 0.0004). This study demonstrated the down-regulation of CRABP1 in the aganglionic hindgut of HSCR, which could provide potential markers or promising new candidate actors for the pathogenesis of HSCR.