Su Lin, Zhang Zhen, Gan Liang, Jiang Qian, Xiao Ping, Zou Jizhen, Li Qi, Jiang Hong
Reproductive Medicine Center, 105 Hospital of People's Liberation Army, Hefei, Anhui, China.
Department of General Surgery, Capital Institute of Pediatrics, Beijing, China.
Exp Mol Pathol. 2016 Oct;101(2):241-248. doi: 10.1016/j.yexmp.2016.09.003. Epub 2016 Sep 13.
Hirschsprung disease (HSCR) is a congenital disorder characterized by the absence of intrinsic ganglion cells in the lower intestine. Genetic factors in the pathogenesis of this disease are under active investigation. As core genes in the planar cell polarity pathway, Celsr3 and Fzd3 are believed to play vital roles in the development of the murine enteric nervous system. The potential association of CELSR3 and FZD3 with the development of HSCR in humans, however, is still unknown. We determined the genotypes of eight CELSR3 and FZD3 polymorphisms in 113 patients. Furthermore, target gene sequencing was used to search for rare mutations in the planar cell polarity genes. The mRNA and protein expression of CELSR3 and FZD3 were explored in patients with HSCR. Class III β-tubulin in colon tissue samples was examined to elucidate enteric innervation patterns. We observed a significant association between the FZD3 rs17059206 polymorphism and HSCR susceptibility (p<0.001). In addition, five rare mutations in CELSR3 were identified in six patients with HSCR. Upregulation of CELSR3 mRNA expression was detected in 80% of aganglionic segments; a similar increase was found for FZD3 protein expression in 81.8% of aganglionic tissues, compared with the ganglionic segments. Immunohistochemical staining on tissue sections revealed obvious excess expression of both molecules in the mucosal layer. The neurite patterns were highly disorganized in the aganglionic bowel segments, with a marked reduction in the prominence of TUJ1 bundles in number, thickness, and length. Our results showed that deregulation of the planar cell polarity genes CELSR3 and FZD3 might disrupt the enteric innervation pattern and consequently contribute to the susceptibility to HSCR.
先天性巨结肠（HSCR）是一种先天性疾病，其特征是下肠道缺乏内在神经节细胞。该疾病发病机制中的遗传因素正在积极研究中。作为平面细胞极性通路中的核心基因，Celsr3和Fzd3被认为在小鼠肠神经系统的发育中起着至关重要的作用。然而，CELSR3和FZD3与人类HSCR发生之间的潜在关联仍不清楚。我们确定了113例患者中8个CELSR3和FZD3多态性的基因型。此外，使用靶基因测序来寻找平面细胞极性基因中的罕见突变。在HSCR患者中探索了CELSR3和FZD3的mRNA和蛋白质表达。检查结肠组织样本中的III类β-微管蛋白以阐明肠神经支配模式。我们观察到FZD3 rs17059206多态性与HSCR易感性之间存在显著关联（p<0.001）。此外，在6例HSCR患者中鉴定出CELSR3的5个罕见突变。在80%的无神经节段中检测到CELSR3 mRNA表达上调；与神经节段相比，在81.8%的无神经节组织中发现FZD3蛋白表达有类似增加。组织切片的免疫组织化学染色显示这两种分子在粘膜层均有明显过度表达。无神经节肠段的神经突模式高度紊乱，TUJ1束在数量、厚度和长度上的突出程度明显降低。我们的结果表明，平面细胞极性基因CELSR3和FZD3的失调可能会破坏肠神经支配模式，从而导致HSCR易感性增加。
