Paneth cell heterogeneity in infection and inflammation

项目总结 克罗恩病(CD)是IBD的一种形式，通常累及小肠，大多数潘氏细胞 住下来。由于大量CD患者对标准治疗没有反应或无效，因此有一种 基于新的作用机制的新的治疗选择的未得到满足的需求。最近的研究表明， 潘氏细胞缺陷可能在回肠和回盲部CD的炎症发生中起关键作用。帕内斯 CD患者或小鼠IBD模型中的细胞通常表现出颗粒表型，最初的特征是Ta博士- 清流，在那里几个Paneth细胞富含抗菌因子，而不是被包装成分泌物 颗粒散布在细胞质中。与内质网缺陷应激、自噬和分泌有关的假说 已经开发出解释这些CD Paneth细胞缺陷的方法，然而因果关系 仍然鲜为人知。到目前为止，现有的研究一直将潘斯细胞视为同质群体。基座 在新发展的时间控制的Paneth细胞报告小鼠模型上，我们建立了单细胞 回肠潘氏细胞在动态平衡、生物失调、感染和炎症条件下的转录图谱。我们 发现潘氏细胞的异质性随着微生物区系的改变、感染、 以及潜在的遗传风险因素。分析不同条件下的Paneth细胞群体揭示了 免疫激活的潘氏细胞亚群，具有独特的抗菌肽特征，丰富的细胞因子 受体，并增强先天免疫和脱颗粒活性。携带签名的Paneth细胞 在回肠CD和UC化生病变中发现了标志物。我们将检验这样的假设：一个动态的潘氏细胞 由遗传、肠道微生物区系和环境因素(如病原体)相互作用调节的异质性 感染是激活潘氏细胞介导的抗炎性诱导和天然保护的关键 更严重的。目标1将通过检查这些潘氏细胞的功能和疾病相关性来确定它们的功能和疾病相关性 天然免疫功能、脱颗粒能力、染色质和表观遗传谱，在体内的作用 分泌的粘膜五肽，与CD Paneth细胞缺陷和临床结果相关。Aim 2将测试如何 两条潘氏细胞内在途径和CD非生物微生物区系通过执行以下步骤影响潘氏细胞异质性 Paneth细胞特异的基因消融和过表达，以及新生的粪便微生物组移植 培育出无菌的潘氏细胞报告小鼠。这一提议解决了一个重大而创新的假设。 与Paneth细胞异质性有关，这可能有助于Paneth细胞驱动的因果关系和机制 CD发病机制。一种调节潘氏细胞激活和炎症保护的新机制 如果被确认，它可能被用于IBD干预。

PROJECT SUMMARY Crohn’s disease (CD) is a form of IBD that commonly involves small intestine, where the majority of Paneth cells reside. As a great number of CD patients show no response or refractory to standard treatment, there is an unmet need for new therapeutic options based on new mechanisms of action. Recent studies suggested that defective Paneth cells may play the key role in initiating inflammation in ileal, and maybe ileocecal CD. Paneth cells in CD patients or mouse IBD models often exhibit granule phenotypes characterized initially by Dr. Ta- Chiang Liu, where several Paneth cell-enriched antimicrobial factors, instead of being packed into secretory granules, are dispersed to the cytoplasm. Hypotheses relating to defective ER stress, autophagy, and secretion have been developed to interpret these CD Paneth cell defects, however the cause-and-effect relationship remains obscure. To date, existing studies has been treating Paneth cells as a homogeneous population. Based on a newly developed temporally controlled Paneth cell reporter mouse model, we establish single cell transcriptomic maps for ileal Paneth cells in homeostasis, dysbiosis, infection and inflammation conditions. We found that there is a profound change in Paneth cell heterogeneity in response to microbiota alteration, infection, and potentially genetic risk factors. Analyzing Paneth cell populations across different conditions revealed an immunologically activated Paneth cell subset that has distinct antimicrobial peptide profile, enriched cytokine receptor, and heightened innate immune and degranulation activities. Paneth cells carrying the signature markers are found in ileal CD and UC metaplastic lesions. We will test the hypothesis that a dynamic Paneth cell heterogeneity regulated by interactions of genetic, gut microbiota, and environmental factors such as pathogen infection is critical for activation of Paneth cell-mediated innate protection against inflammatory induction and aggravation. Aim 1 will determine the function and disease relevance of these Paneth cells by examine their innate immune function, degranulation capacity, chromatin and epigenetic profile, in vivo role of specifically secreted mucosal pentraxins, and relevance to CD Paneth cell defects and clinical outcomes. Aim 2 will test how two Paneth cell intrinsic pathways and CD dysbiotic microbiota affect Paneth cell heterogeneity, by performing Paneth cell specific genetic ablation and overexpression, as well as fecal microbiome transplantation in newly developed germ-free Paneth cell reporter mice. This proposal addresses a significant and innovative hypothesis relating to Paneth cell heterogeneity that may contribute to the causality and mechanism of Paneth cell-driven CD pathogenesis. Once a new mechanism regulating proper Paneth cell activation and inflammatory protection is identified, it may be leveraged for IBD intervention.