Quantitative Proteomics Reveals Association of Neuron Projection Development Genes ARF4, KIF5B, and RAB8A With Hirschsprung Disease.
Quantitative Proteomics Reveals Association of Neuron Projection Development Genes ARF4, KIF5B, and RAB8A With Hirschsprung Disease.
复制标题
定量蛋白质组学揭示神经元投射发育基因 ARF4、KIF5B 和 RAB8A 与先天性巨结肠的关联
DOI：
10.1074/mcp.ra120.002325
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Jiang Q
中科院分区：
文献类型：
--
作者：
Zhang Q;Wu L;Bai B;Li D;Xiao P;Li Q;Zhang Z;Wang H;Li L;Jiang Q
先天性巨结肠是一组异质性的神经嵴病，其特征是沿着不同长度的肠沿着缺乏肠神经节。遗传缺陷在HSCR的发病机制中起着重要作用，而所有已知基因（位点）中致病性变体的家族研究仅表明不完全的突变和未知原因的可变表达。在这里，我们应用大规模的，定量的蛋白质组学的人类结肠组织从21例患者使用同量异位素标签的相对和绝对定量。方法采用生物信息学分析。通过平行反应监测验证证实了选定的结果。最后通过Western blot对差异表达蛋白进行鉴定。共鉴定了5341种人结肠组织蛋白质。其中，664个差异大于1.2倍的蛋白质在6组中被鉴定：A1和A2组来自于长节段HSCR患者的神经节细胞和无神经节细胞结肠的蛋白质（n = 7），B1和B2组来自于短节段HSCR患者的神经节细胞和无神经节细胞结肠的蛋白质（n = 7），B1和B2组来自于短节段HSCR患者的神经节细胞和无神经节细胞结肠的蛋白质（n = 7）。C1和C2组汇集来自女性短节段HSCR患者（n = 7）的神经节和无神经节结肠的蛋白质。在此基础上，从核糖体、胞吞、剪接体、氧化磷酸化和细胞粘附等5个途径中选取49个蛋白质进行平行反应监测验证。在15个配对的结肠样本中，使用Western blot验证了结肠无神经节部分中三个神经元投射发育基因ARF4、KIF5B和RAB8A的下调。本研究的发现将为HSCR的发病机制提供新的线索，并促进治疗靶点的开发。
Hirschsprung disease (HSCR) is a heterogeneous group of neurocristopathy characterized by the absence of the enteric ganglia along a variable length of the intestine. Genetic defects play a major role in the pathogenesis of HSCR, whereas family studies of pathogenic variants in all the known genes (loci) only demonstrate incomplete penetrance and variable expressivity for unknown reasons. Here, we applied large-scale, quantitative proteomics of human colon tissues from 21 patients using isobaric tags for relative and absolute quantification. method followed by bioinformatics analysis. Selected findings were confirmed by parallel reaction monitoring verification. At last, the interesting differentially expressed proteins were confirmed by Western blot. A total of 5341 proteins in human colon tissues were identified. Among them, 664 proteins with >1.2-fold difference were identified in six groups: groups A1 and A2 pooled protein from the ganglionic and aganglionic colon of male, long-segment HSCR patients (n = 7); groups B1 and B2 pooled protein from the ganglionic and aganglionic colon of male, short-segment HSCR patients (n = 7); and groups C1 and C2 pooled protein from the ganglionic and aganglionic colon of female, short-segment HSCR patients (n = 7). Based on these analyses, 49 proteins from five pathways were selected for parallel reaction monitoring verification, including ribosome, endocytosis, spliceosome, oxidative phosphorylation, and cell adhesion. The downregulation of three neuron projection development genes ARF4, KIF5B, and RAB8A in the aganglionic part of the colon was verified in 15 paired colon samples using Western blot. The findings of this study will shed new light on the pathogenesis of HSCR and facilitate the development of therapeutic targets.