Paneth cell heterogeneity in infection and inflammation

项目概要 克罗恩病 (CD) 是 IBD 的一种形式，通常累及小肠，其中大多数潘氏细胞 居住。由于大量 CD 患者对标准治疗没有反应或难治，因此存在一种 对基于新作用机制的新治疗方案的需求未得到满足。最近的研究表明 有缺陷的潘氏细胞可能在回肠炎症甚至回盲肠 CD 的引发中起关键作用。帕内斯 CD 患者或小鼠 IBD 模型中的细胞通常表现出最初由 Ta 博士表征的颗粒表型- 蒋刘，其中几种富含潘氏细胞的抗菌因子，而不是被包装到分泌物中 颗粒，分散到细胞质中。与内质网应激、自噬和分泌缺陷相关的假设 已被开发来解释这些 CD 潘氏细胞缺陷，但是因果关系 仍然晦涩难懂。迄今为止，现有的研究一直将潘氏细胞视为同质群体。基于 在新开发的时间控制潘氏细胞报告小鼠模型上，我们建立了单细胞 回肠潘氏细胞在稳态、生态失调、感染和炎症条件下的转录组图谱。我们 发现潘氏细胞异质性因微生物群改变、感染、 以及潜在的遗传风险因素。分析不同条件下的潘氏细胞群揭示了 免疫激活的潘氏细胞亚群，具有独特的抗菌肽谱、丰富的细胞因子 受体，并增强先天免疫和脱粒活性。潘氏细胞携带签名 标记物见于回肠 CD 和 UC 化生病变。我们将检验动态潘氏池的假设 异质性受遗传、肠道微生物群和病原体等环境因素相互作用的调节 感染对于激活潘氏细胞介导的针对炎症诱导的先天保护至关重要 加重。目标 1 将通过检查这些潘氏细胞来确定其功能和疾病相关性 先天免疫功能、脱颗粒能力、染色质和表观遗传特征、特定的体内作用 分泌的粘膜五聚蛋白，以及与 CD 潘氏细胞缺陷和临床结果的相关性。目标 2 将测试如何 两条潘氏细胞内在途径和CD失调微生物群影响潘氏细胞异质性，通过执行 潘氏细胞特异性基因消融和过度表达，以及新的粪便微生物组移植 开发出无菌潘氏细胞报告小鼠。该提案提出了一个重要且创新的假设 与潘氏细胞异质性相关，可能有助于潘氏细胞驱动的因果关系和机制 CD发病机制。一旦出现调节潘氏细胞适当激活和炎症保护的新机制 一旦确定，它可用于 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.