RNA-Seq of human sorted colonic macrophages from Hirschsprung's disease patients

Hirschsprung's disease (HD) is a congenital intestinal motility disorder defined by the absence of enteric ganglia cells (aganglionose) in the distal colon and characterized by bowel obstruction and the formation of a megacolon. Although, routinely treated by surgical removal of aganglionic segments the most life-threatening complication is the development of HD-associated enterocolitis with still unknown pathogenesis. Due to the lack of enteric ganglia cells HD patients show extramural innervation of acetylcholine-secreting (cholinergic) nerve fibers. Cholinergic signals have been reported to control excessive inflammatory immune responses, however, the impact on the colonic microbiom and enterocolitis manifestation in HD patients is unknown. We grouped HD patients according to their degree of mucosal cholinergic innervation in the distal colon into fiber-high and fiber-low patients. Colonic mucosal macrophages from fiber-high and fiber-low as well as muscularis macrophages from HD patients were FACS-sorted as viable CD45+HLA-DR+CD64+ cells and RNA-Seq performed. Blood-derived in vitro differentiated M1 and M2 macrophages from healthy adult donor were included as controls. These data will clarify how the presence of cholinergic neurons affect the phenotype of colonic macrophages.

先天性巨结肠（Hirschsprung's disease, HD）是一类先天性肠动力障碍性疾病，以结肠远端缺乏肠神经节细胞（无神经节症，aganglionosis）为特征，临床表现为肠梗阻及巨结肠形成。尽管该病常规治疗手段为手术切除无神经节肠段，但其最致命的并发症为HD相关性小肠结肠炎，该并发症的发病机制至今尚未明确。由于肠神经节细胞缺失，HD患者的结肠壁外存在分泌乙酰胆碱的胆碱能（cholinergic）神经纤维支配。已有研究证实胆碱能信号可调控过度炎症免疫应答，但此类信号对HD患者结肠微生物组及HD相关性小肠结肠炎发病的影响仍不清晰。本研究根据HD患者结肠远端黏膜的胆碱能神经支配程度，将其分为高纤维组与低纤维组；通过荧光激活细胞分选（Fluorescence-Activated Cell Sorting, FACS）从两组患者中分离得到结肠黏膜巨噬细胞及肌层巨噬细胞，分选标记为活细胞CD45+HLA-DR+CD64+，并对其开展RNA测序（RNA-Seq）。同时纳入健康成人供体来源的体外诱导分化M1、M2型巨噬细胞作为对照。本数据集旨在阐明胆碱能神经元的存在如何影响结肠巨噬细胞的表型特征。