Senger Stefania, Ingano Laura, Freire Rachel, Anselmo Antony, Zhu Weishu, Sadreyev Ruslan, Walker William Allan, Fasano Alessio
Department of Pediatrics, Mucosal Immunology and Biology Research Center, Massachusetts General Hospital, Boston, Massachusetts.
Harvard Medical School, Boston, Massachusetts.
Cell Mol Gastroenterol Hepatol. 2018 Jan 31;5(4):549-568. doi: 10.1016/j.jcmgh.2018.01.014. eCollection 2018.
BACKGROUND & AIMS: Untreated necrotizing enterocolitis (NEC) can lead to massive inflammation resulting in intestinal necrosis with a high mortality rate in preterm infants. Limited access to human samples and relevant experimental models have hampered progress in NEC pathogenesis. Earlier evidence has suggested that bacterial colonization of an immature and developing intestine can lead to an abnormally high inflammatory response to bacterial bioproducts. The aim of our study was to use human fetal organoids to gain insights into NEC pathogenesis.
RNA sequencing analysis was performed to compare patterns of gene expression in human fetal-derived enterospheres (FEnS) and adult-derived enterospheres (AEnS). Differentially expressed genes were analyzed using computational techniques for dimensional reduction, clustering, and gene set enrichment. Unsupervised cluster analysis, Gene Ontology, and gene pathway analysis were used to predict differences between gene expression of samples. Cell monolayers derived from FEnS and AEnS were evaluated for epithelium function and responsiveness to lipopolysaccharide and commensal bacteria.
Based on gene expression patterns, FEnS clustered according to their developmental age in 2 distinct groups: early and late FEnS, with the latter more closely resembling AEnS. Genes involved in maturation, gut barrier function, and innate immunity were responsible for these differences. FEnS-derived monolayers exposed to either lipopolysaccharide or commensal showed that late FEnS activated gene expression of key inflammatory cytokines, whereas early FEnS monolayers did not, owing to decreased expression of nuclear factor-κB-associated machinery.
Our results provide insights into processes underlying human intestinal development and support the use of FEnS as a relevant human preclinical model for NEC. Accession number of repository for expression data: GSE101531.
未经治疗的坏死性小肠结肠炎（NEC）可导致严重炎症，进而引起早产婴儿肠道坏死，死亡率很高。获取人类样本和相关实验模型的机会有限，阻碍了NEC发病机制研究的进展。早期证据表明，未成熟且正在发育的肠道细菌定植可导致对细菌生物产物的异常高炎症反应。我们研究的目的是利用人类胎儿类器官深入了解NEC的发病机制。
进行RNA测序分析，以比较人类胎儿来源的肠球（FEnS）和成人来源的肠球（AEnS）中的基因表达模式。使用计算技术进行降维、聚类和基因集富集分析差异表达基因。采用无监督聚类分析、基因本体论和基因通路分析来预测样本基因表达之间的差异。评估来自FEnS和AEnS的细胞单层的上皮功能以及对脂多糖和共生细菌的反应性。
根据基因表达模式，FEnS根据其发育年龄分为2个不同的组：早期FEnS和晚期FEnS，后者与AEnS更相似。参与成熟、肠道屏障功能和先天免疫的基因导致了这些差异。暴露于脂多糖或共生细菌的FEnS来源的单层细胞显示，晚期FEnS激活了关键炎症细胞因子的基因表达，而早期FEnS单层细胞未激活，这是由于核因子κB相关机制的表达降低。
我们的结果为人类肠道发育的潜在过程提供了见解，并支持将FEnS用作NEC相关的人类临床前模型。表达数据储存库登录号：GSE101531。
