Gastroenterology最新文献

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A PERSONALIZED HUMAN PANETH CELL MODEL THAT IS RESPONSIVE TO MICROBES/MICROBIAL LIGANDS 一个对微生物/微生物配体有反应的个性化人类平板细胞模型

IF 29.4 1区医学 Q1 GASTROENTEROLOGY & HEPATOLOGY

Gastroenterology

Pub Date : 2026-01-22 DOI: 10.1053/j.gastro.2025.10.082

Shachi Patel, Monica Silveira Wagner, Olivia Bay, Christian Wong Valencia, Eliska Zgarbova, Cynthia Rodriguez, Daniel Leal, Michifumi Yamashita, Suzanne Devkota, Kathrin Michelsen, Stephan Targan, Robert Barrett

Section snippets

Methods

iPSCs from a control individual were directed to HIOs and then dissociated to generate purified populations of epithelial only-HIOs (eHIOs). eHIOs were passaged weekly in proliferation medium (EGF, Noggin and CHIR99021) and the γ-secretase inhibitor, DAPT, was added to direct towards a Paneth cell fate. Flow cytometry, qPCR and immunocytochemistry were used to estimate Paneth cell number, gene expression and presence of various AMPs respectively. To confirm the functionality of this cell type,

Results

iPSC-derived eHIOs could be maintained for at least 4 months in proliferation medium. Flow cytometry analysis of the Paneth marker lysozyme revealed that the population of Paneth cells in eHIOs significantly increased from ∼1% in proliferation media to ∼30% upon treatment with DAPT. qPCR analysis demonstrated that DAPT treatment significantly increased the expression of the Paneth cell associated genes DEFA5, DEFA6, ITLN2, REG3A and PLA2G2A. Immunoctyochemistry revealed that DAPT treated eHIOs

Conclusion

We have successfully developed a methodology to enrich Paneth cells in iPSC-derived HIOs. Given that iPSCs can be generated from donor cells stored in well characterized biorepositories or from a small blood draw from any CD patient, this modeling system now opens a new avenue of research by allowing an examination of how environmental factors (microbes/cytokines) and/or genetic variations influence human Paneth cell function in a personalized manner.

. qPCR analysis of Paneth cell related genes

方法将来自对照个体的sipscs导入HIOs，然后分离生成纯化的上皮型HIOs （eHIOs）群体。eHIOs在增殖培养基（EGF， Noggin和CHIR99021）中每周传代，并加入γ-分泌酶抑制剂DAPT来指导Paneth细胞的命运。采用流式细胞术、qPCR和免疫细胞化学分别检测Paneth细胞数量、基因表达和各种amp的存在。结果sipsc衍生的eHIOs可在增殖培养基中维持至少4个月。Paneth标记溶菌酶的流式细胞术分析显示，DAPT处理后，eHIOs中Paneth细胞的数量从增殖培养基中的~ 1%显著增加到~ 30%。qPCR分析显示，DAPT处理显著增加了Paneth细胞相关基因DEFA5、DEFA6、ITLN2、REG3A和PLA2G2A的表达。免疫细胞化学显示DAPT对ehios有治疗作用。结论我们已经成功开发了一种方法，可以在ipsc来源的HIOs中富集Paneth细胞。鉴于iPSCs可以从储存在特征良好的生物储存库中的供体细胞或从任何CD患者的少量血液中生成，该建模系统现在通过允许检查环境因素（微生物/细胞因子）和/或遗传变异如何以个性化的方式影响人类Paneth细胞功能，开辟了新的研究途径。Paneth细胞相关基因的qPCR分析

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引用次数: 0

PEOPLE WITH ULCERATIVE COLITIS HAVE ALTERED EPITHELIAL PURINERGIC P2 RECEPTOR EXPRESSION AND FUNCTION 溃疡性结肠炎患者上皮嘌呤能p2受体表达和功能发生改变

IF 29.4 1区医学 Q1 GASTROENTEROLOGY & HEPATOLOGY

Gastroenterology

Pub Date : 2026-01-22 DOI: 10.1053/j.gastro.2025.10.139

Ana Pettijohn, Kayla McGary, Kristen Engevik

Section snippets

Background

Purines are well-established as potent signaling molecules that regulate cellular homeostasis through activation of purinergic receptors. There are 19 distinct purinergic receptor subtypes, classified as P1 and P2 receptors. While purinergic receptors are known to play important roles in immune cell responses during inflammation and infection, their function in the intestinal epithelium, especially in inflammatory bowel disease (IBD), remains largely unknown. We hypothesized that key purinergic

Methods & Results

We queried single cell RNA sequencing data from the Human Protein Atlas and CZ CELLxGENE Discover databases to assess purinergic receptor distribution in healthy human colon. In silico analysis revealed limited P1 receptor expression in the intestinal epithelium but robust expression in immune cell types. By contrast, healthy small and large intestine epithelium expressed the P2 receptors P2RY1, P2RY2, and P2RX4 across nearly all epithelial cell types, while immune cells were enriched for

Conclusions

These findings suggest an imbalance in epithelial and immune cell purinergic signaling in the setting of ulcerative colitis and point to unique roles of purinergic signaling with the intestine.

背景嘌呤被认为是一种有效的信号分子，通过激活嘌呤能受体来调节细胞稳态。有19种不同的嘌呤能受体亚型，分为P1和P2受体。虽然已知嘌呤能受体在炎症和感染期间的免疫细胞反应中发挥重要作用，但它们在肠上皮中的功能，特别是在炎症性肠病（IBD）中的功能仍不清楚。我们查询了来自人类蛋白图谱和CZ CELLxGENE Discover数据库的单细胞RNA测序数据，以评估健康人类结肠中嘌呤能受体的分布。芯片分析显示P1受体在肠上皮中表达有限，但在免疫细胞类型中表达强劲。相比之下，健康小肠和大肠上皮在几乎所有上皮细胞类型中表达P2受体P2RY1、P2RY2和P2RX4，而免疫细胞则表达P2受体P2RY1、P2RY2和P2RX4。结论这些发现提示溃疡性结肠炎背景下上皮和免疫细胞嘌呤能信号的不平衡，并指出嘌呤能信号在肠道中的独特作用。

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引用次数: 0

ABNORMAL PRE-OPERATIVE ANORECTAL MANOMETRY IS ASSOCIATED WITH INFLAMMATION OF THE RECTAL CUFF IN IPAA PATIENTS BUT NOT ENDOSCOPIC INFLAMMATORY POUCH DISEASE 术前肛门直肠测压异常与ipaa患者的直肠袖带炎症有关，而与内窥镜炎性袋病无关