Division of Pediatric Surgery, Department of Surgery, University of Tennessee Health Sciences Center, Memphis, TN, USA.
Department of Surgery, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA.
FASEB J. 2020 Aug;34(8):10931-10947. doi: 10.1096/fj.202000699R. Epub 2020 Jun 27.
Enteric nervous system (ENS) development is governed by interactions between neural crest cells (NCC) and the extracellular matrix (ECM). Hirschsprung disease (HSCR) results from incomplete NCC migration and failure to form an appropriate ENS. Prior studies implicate abnormal ECM in NCC migration failure. We performed a comparative microarray of the embryonic distal hindgut of wild-type and EdnrB mice that model HSCR and identified laminin-β1 as upregulated in EdnrB colon. We identified decreased expression of 37/67 kDa laminin receptor (LAMR), which binds laminin-β1, in human HSCR myenteric plexus and EdnrB NCC. Using a combination of in vitro gut slice cultures and ex vivo organ cultures, we determined the mechanistic role of LAMR in NCC migration. We found that enteric NCC express LAMR, which is downregulated in human and murine HSCR. Binding of LAMR by the laminin-β1 analog YIGSR promotes NCC migration. Silencing of LAMR abrogated these effects. Finally, applying YIGSR to E13.5 EdnrB colon explants resulted in 80%-100% colonization of the hindgut. This study adds LAMR to the large list of receptors through which NCC interact with their environment during ENS development. These results should be used to inform ongoing integrative, regenerative medicine approaches to HSCR.
肠神经系统（ENS）的发育受神经嵴细胞（NCC）与细胞外基质（ECM）之间相互作用的调控。先天性巨结肠（HSCR）是由于 NCC 迁移不完全和不能形成适当的 ENS 所致。先前的研究表明 ECM 在 NCC 迁移失败中起作用。我们对野生型和 EdnrB 小鼠胚胎远端后肠进行了比较微阵列分析，EdnrB 小鼠可模拟 HSCR，并发现层粘连蛋白-β1 在 EdnrB 结肠中上调。我们发现，人 HSCR 肌间神经丛和 EdnrB NCC 中 37/67 kDa 层粘连蛋白受体（LAMR）的表达减少，LAMR 结合层粘连蛋白-β1。通过体外肠切片培养和离体器官培养的组合，我们确定了 LAMR 在 NCC 迁移中的作用机制。我们发现肠 NCC 表达 LAMR，而人 HSCR 和鼠 HSCR 中 LAMR 的表达下调。层粘连蛋白-β1 类似物 YIGSR 与 LAMR 的结合促进了 NCC 的迁移。LAMR 的沉默消除了这些作用。最后，将 YIGSR 应用于 E13.5 EdnrB 结肠外植体，导致后肠 80%-100%的定植。这项研究将 LAMR 添加到 NCC 在 ENS 发育过程中与环境相互作用的大量受体中。这些结果应被用于告知正在进行的综合、再生医学方法来治疗 HSCR。
