Innate enteric immunity during induced Paneth cell deficiency

诱导潘氏细胞缺陷期间的先天肠道免疫

• 批准号: 8493004
• 负责人: Andre J. Ouellette
• 金额: $ 19.27万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-15 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8493004
• 关键词: Acute; Affect; Age; Autophagocytosis; Bacteria; Bacterial Translocation; Bacteroidetes; Binding; Cell Count; Cell Differentiation process; Cell Lineage; Cell Maintenance; Cell Proliferation; Chronic; Complex; Defensins; Development; E1A-associated p300 protein; EP300 gene; Embryo; Enteral; Enterobacteriaceae; Epithelial; Eubacterium; Functional disorder; Future; Genetic Transcription; Growth; Health; Homeostasis; Host Defense; Immunity; Immunology; Impairment; Infection; Infiltration; Inflammation; Intestines; Lactobacillus; Life; Lymphoid Cell; Maintenance; Mediating; Mining; Mole the mammal; Monitor; Mucosal Immunity; Mus; Mutation; Myeloid Cells; Oral; Organoids; Paneth Cells; Phenotype; Predisposition; Recovery; Research; Research Proposals; Role; Salmonella; Secretory Vesicles; Signal Pathway; Signal Transduction; Small Intestines; Stem cells; Stress; Testing; Time; Transcription Coactivator; base; dosage; drug withdrawal; in vivo; inflammatory marker; innovation; intestinal crypt; intestinal epithelium; oral infection; pathogenic bacteria; progenitor; programs; public health relevance; response; restoration; small molecule

DESCRIPTION (provided by applicant): The proposed studies will test the hypothesis that induced imbalance in Wnt/¿-catenin signaling results in Paneth cell depletion, dysbiosis of the resident microbiota, inflammation, enhanced susceptibility to infection by pathogenic bacteria, and that these effects can be reversed or ameliorated using a specific modulator of the Wnt/¿-catenin signaling pathway. Anatomically restricted to the small bowel, Paneth cells release ¿-defensins and host defense molecules, and maintenance of Paneth cell homeostasis is essential to small intestinal health and integrity. The Wnt signaling cascade has a critical role i gut ontogeny, and Wnt signaling mediated by ¿- catenin p300 coactivator complexes is critical for Paneth cell differentiation. ¿-catenin/CBP-mediated transcription maintains stem cell potency; in contrast, ¿-catenin/p300 transcription complexes initiate differentiation programs of more limited proliferative capacity. The primary ¿-catenin co-activators CBP and p300 share extensive similarity but interact with ¿-catenin via their distinctive N termini, which has enabled development of small molecule antagonists that specifically inhibit their association with ¿-catenin and participation in ¿-catenin-mediated transcription. For example, ICG-001 promotes Wnt-dependent differentiation by binding to CBP specifically, but not to p300, blocking CBP/¿-catenin transcription without 1) interfering with p300, 2) altering levels of ¿-catenin, but still ) allowing CBP to participate in other transcription complexes. Conversely, IQ-1 antagonizes the ¿-catenin-p300 interaction but not CBP associations, providing a means for inducing selective Paneth cell deficiency. Thus, we will test the hypothesis that antagonism of ¿- catenin/p300 signaling in mice with IQ-1 induces Paneth cell depletion and dysfunction. The impact of disrupting Paneth cell homeostasis will be determined by monitoring the time course of IQ-1 induced Paneth cell deficiency and coincident changes in expression of Paneth cell specific markers and induced inflammation evident as infiltration and translocation of myeloid and lymphoid cells. Effects of Paneth cell deficiency on the composition of the prokaryotic and eukaryotic microbiota and on barrier integrity inferred from translocation of gut bacteria in vivo will be characterized. After establishing Paneth cell deficiency, the effects of drug withdrawal and of ICG-001 administration to correct co-activator imbalance and restore Paneth cell differentiation and barrier integrity will be determined, and Paneth cell autonomous responses to IQ-1 and ICG- 001 will be determined on crypt organoids ex vivo. The effect of reestablishing Paneth cell lineage allocation and crypt homeostasis on reversing prokaryotic and eukaryotic dysbiosis also will be determined. We anticipate that IQ-1-induced Paneth cell depletion will evoke proinflammatory responses, induce dysbiosis, and enhance susceptibility to bacterial translocation and that ICG-001 will selectively enable p300/¿-catenin driven transcription to promote Wnt-dependent Paneth cell differentiation and accelerate recovery of small bowel homeostasis.

描述（应用程序提供）：拟议的研究将检验以下假说：诱导Wnt/� -Catenin信号传导导致Paneth细胞部署，居民微生物群的营养不良，感染，增强的病原细菌感染的易感性，并使用特定的模型来逆转或舒余。在解剖学上仅限于小肠，泛细胞释放�-防御素和宿主防御分子，维持Paneth细胞体内平衡对于小肠道健康和完整性至关重要。 Wnt信号传导级联反应具有至关重要的作用，并且由Catenin P300共激活因子复合物介导的Wnt信号传导对于Paneth细胞分化至关重要。 �-蛋白酶/CBP介导的转录保持干细胞效力；相反，� -Catenin/p300转录复合物启动具有更有限的增殖能力的分化程序。主要的� -Catenin共激活剂CBP和P300具有广泛的相似性，但通过其独特的N末端与� -Catenin相互作用，这已启用 小分子拮抗剂的发展，这些拮抗剂特别抑制了它们与� -catenin的关联并参与 - - 卡宁蛋白介导的转录。例如，ICG -001通过与CBP结合而不是与P300结合来促进Wnt依赖性分化，从而阻止CBP/¿-CATENIN转录，而无需1）干扰P300，2）改变 - Catenin的水平，但仍然存在），但仍然存在）允许CBP参与其他转录复合物。相反，IQ-1拮抗了-Catenin-P300相互作用，而不是CBP关联，为诱导的Paneth细胞缺乏提供了一种手段。这是，我们将检验以下假设：与IQ -1的小鼠中catenin/p300信号的拮抗作用会诱导Paneth细胞的部署和功能障碍。破坏Paneth细胞体内平衡的影响将通过监测IQ-1诱导的Paneth细胞缺乏症的时间过程和Paneth细胞特异性标记表达的同时变化，并引起感染证据，因为髓样和淋巴样细胞的浸润和易位。泛细胞缺乏对原核和真核微生物群组成以及体内肠道细菌易位推断的屏障完整性的影响。在建立了泛细胞缺乏症之后，将确定药物戒断和ICG-001给药对校正共激活器失衡的影响，并恢复Paneth细胞分化和屏障完整性，并且将在Crypto tone上确定Paneth细胞对IQ-1和ICG-001的自主反应。还将确定重新建立Paneth细胞谱系分配和加密稳态对逆转原核和真核营养不良的影响。我们预计，IQ-1引起的Paneth细胞部署将引起促炎反应，诱发营养不良并增强对细菌易位的易感性，并且ICG-001将有选择地启用P300/�-Catenin驱动的转录，以促进Wnt依赖的Paneth Cell依赖的Paneth Cell Eccessiation和Acceltiation respeconles Recuthere Recuceprome couseconlese recousplerate smellow Bowelelatorestersiss。