Windster Jonathan D, Kakiailatu Naomi J M, Kuil Laura E, Antanaviciute Agne, Sacchetti Andrea, MacKenzie Katherine C, Peulen-Zink Joke, Kan Tsung W, Bindels Eric, de Pater Emma, Doukas Michail, van den Bosch Thierry P P, Yousefi Soheil, Barakat Tahsin-Stefan, Meeussen Conny J H M, Sloots Pim C E J, Wijnen Rene M H, Parikh Kaushal, Boesmans Werend, Melotte Veerle, Hofstra Robert M W, Simmons Alison, Alves Maria M
Department of Clinical Genetics, Erasmus University Medical Center, Sophia Children's Hospital, Rotterdam, the Netherlands; Department of Pediatric Surgery, Erasmus University Medical Center, Sophia Children's Hospital, Rotterdam, the Netherlands.
Department of Clinical Genetics, Erasmus University Medical Center, Sophia Children's Hospital, Rotterdam, the Netherlands.
Gastroenterology. 2025 May;168(5):965-979.e12. doi: 10.1053/j.gastro.2024.12.011. Epub 2024 Dec 24.
BACKGROUND & AIMS: The enteric nervous system (ENS), which is composed of neurons and glia, regulates intestinal motility. Hirschsprung disease (HSCR) results from defects in ENS formation; however, although neuronal aspects have been studied extensively, enteric glia remain disregarded. This study aimed to explore enteric glia diversity in health and disease.
Full-thickness intestinal resection material from pediatric controls and patients with HSCR was collected, dissociated, and enriched for the ENS population through fluorescence-activated cell sorting. Single-cell RNA sequencing was performed to uncover the transcriptomic diversity of the ENS in controls and HSCR patients, as well as in wild-type and ret mutant zebrafish. Immunofluorescence and fluorescence in situ hybridization confirmed the presence of distinct subtypes.
Two major enteric glial classes emerged in the pediatric intestine: Schwann-like enteric glia, which are reminiscent of Schwann cells, and enteric glia expressing classical glial markers. Comparative analysis with previously published datasets confirmed our classification and revealed that although classical enteric glia are predominant prenatally, Schwann-like enteric glia become more abundant postnatally. In HSCR, ganglionic segments mirrored controls and aganglionic segments featured only Schwann-like enteric glia. Leveraging the regenerative potential of Schwann cells, we explored therapeutic options using a ret mutant zebrafish. Prucalopride, a serotonin-receptor (5-HT) agonist, induced neurogenesis partially rescuing the HSCR phenotype in ret mutants.
Two major enteric glial classes were identified in the pediatric intestine, highlighting the significant postnatal contribution of Schwann-like enteric glia to glial heterogeneity. Crucially, these glial subtypes persist in aganglionic segments of patients with HSCR, offering a new target for their treatment using 5-HT agonists.
由神经元和神经胶质细胞组成的肠神经系统（ENS）调节肠道蠕动。先天性巨结肠症（HSCR）是由ENS形成缺陷导致的；然而，尽管神经元方面已得到广泛研究，但肠神经胶质细胞仍被忽视。本研究旨在探索健康和疾病状态下肠神经胶质细胞的多样性。
收集来自儿科对照患者和HSCR患者的全层肠切除材料，解离后通过荧光激活细胞分选富集ENS群体。进行单细胞RNA测序以揭示对照患者和HSCR患者以及野生型和ret突变斑马鱼中ENS的转录组多样性。免疫荧光和荧光原位杂交证实了不同亚型的存在。
在小儿肠道中出现了两种主要的肠神经胶质细胞类型：类似于施万细胞的施万样肠神经胶质细胞，以及表达经典神经胶质细胞标志物的肠神经胶质细胞。与先前发表的数据集进行比较分析证实了我们的分类，并表明尽管经典肠神经胶质细胞在产前占主导地位，但施万样肠神经胶质细胞在出生后变得更加丰富。在HSCR中，神经节段与对照相似，无神经节段仅具有施万样肠神经胶质细胞。利用施万细胞的再生潜力，我们使用ret突变斑马鱼探索了治疗方案。普芦卡必利，一种5-羟色胺受体（5-HT）激动剂，诱导神经发生，部分挽救了ret突变体中的HSCR表型。
在小儿肠道中鉴定出两种主要的肠神经胶质细胞类型，突出了施万样肠神经胶质细胞对神经胶质细胞异质性的重要产后贡献。至关重要的是，这些神经胶质细胞亚型在HSCR患者的无神经节段中持续存在，为使用5-HT激动剂治疗提供了新的靶点。
