Role of Interferon lambda signaling in mucosal homeostasis and infantile-onset inflammatory bowel disease

干扰素 lambda 信号传导在粘膜稳态和婴儿发病炎症性肠病中的作用

• 批准号: 10534008
• 负责人: Jodie Ouahed
• 金额: $ 5.4万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-30 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10534008
• 关键词: Age; Award; Cells; Colitis; Cytokine Receptors; Data; Development; Disease; Epithelial; Epithelial Cells; Equipoise; Funding; Gastrointestinal tract structure; Generations; Genetic Predisposition to Disease; Homeostasis; Human; Immune; Immune signaling; In Vitro; Inflammatory Bowel Diseases; Interferon Type I; Interferon-alpha; Interferons; Intestines; Lead; Life; Maternity leave; Mucous Membrane; Mus; Mutation; Organoids; Oxidative Stress; Patients; Physicians; Play; Predisposition; Productivity; Reporting; Research Assistant; Role; Scientist; Signal Transduction; Structure; Technology; Therapeutic; Time; Training; Variant; Viral; Wild Type Mouse; Work; gut inflammation; immune function; improved; in vivo; infancy; intestinal epithelium; microbiome components; microorganism; murine colitis; novel strategies; prevent; response; single-cell RNA sequencing; skills; transcriptome sequencing

PROJECT SUMMARY/ABSTRACT The gastrointestinal tract is composed of a single layer of epithelial cells that is in equipoise with immune cells and a vast number of microorganisms. Inappropriate responses to these microorganisms, either through genetic predisposition, altered immune or epithelial responses, or yet to be defined environmental influences, are postulated to lead to inflammatory bowel diseases (IBD). The immune signals that recognize and respond to bacterial and viral components of the microbiome remain incompletely understood. Interferons (IFNs) play a major role in antiviral immune defense in the intestinal epithelium, and are also important in regulating proliferation, differentiation, survival and effector functions of immune and non-immune cells. There are three classes of IFNs: type I IFNs (IFNα, β, and others), type II IFN (IFN) and type III IFNs, or IFNλs. To date, most studies investigating the use of IFNs on IBD have focused on type I IFNs and were not found to be effective. Dr. Ivan Zanoni reported that IFNλ decreases oxidative stress and intestinal damage in a murine model of colitis and that exogenous IFNλ can suppress intestinal inflammation. Importantly, we identified two unrelated patients with infantile-onset IBD with rare and functionally deleterious mutations in IFNλ2 and IFNλ3. Of note, each patient’s disease improved significantly with age. We have preliminary data that IFNλ2 and IFNλ3 may be more important in the first months of life than FNλ1, and data illustrating more severe murine colitis in Ifnλ3-/- mice as compared to wild type mice. Taken together, we hypothesize that INFλs are essential modulators of mucosal homeostasis, prevent development of IBD, and hold therapeutic potential. Current therapeutics available for the management of IBD fail to treat a large number of patients. This work will provide a better understanding of the role of IFNλ in mucosal homeostasis, and may provide the groundwork to implement novel strategies to treat IBD by manipulating IFNλ signaling. Unraveling the role of IFNλ in maintaining mucosal homeostasis will be achieved through the following aims: (1) Establishing the developmental expression of IFNλs, related cytokines and receptors in humans at different ages using bulk and single cell RNA sequencing technologies (2) Determining the role of IFNλ signaling in predisposition to development of colitis in vivo using various murine models of colitis; (3) Characterizing the functional consequences of patient-encoded IFNλ variants in vitro using T84 cells as well as human control and patient-derived intestinal organoids. The K08 award is instrumental in enabling Dr. Ouahed’s acquisition of the necessary structured training to become proficient in critical skills: RNAseq analysis, murine models of colitis and immune analyses, and generation/manipulation of intestinal organoids and epithelial analyses, complimented with didactic coursework, assisting her path to independence as a successful physician-scientist. The purpose of this supplement is to provide 12 months of funding for a full-time research assistant. This additional support is critical to assure continued productivity of Dr. Ouahed’s work during and immediately following her upcoming maternity leave.

项目摘要/摘要 胃肠道由与免疫细胞平衡的单层上皮细胞组成。 以及大量的微生物。对这些微生物的不适当反应，要么是通过基因 易感性、免疫或上皮性反应改变，或尚未确定的环境影响，是 推测会导致炎症性肠病(IBD)。识别和响应的免疫信号 微生物组的细菌和病毒成分仍然不完全清楚。干扰素(IFN)发挥作用 在肠道上皮细胞的抗病毒免疫防御中起主要作用，也在调节 免疫和非免疫细胞的增殖、分化、存活和效应功能。一共有三个 干扰素类：I型干扰素(干扰素α，β等)、II型干扰素(干扰素)和III型干扰素，或干扰素λS。 研究使用IFN治疗IBD的研究主要集中在I型IFN上，但没有发现有效。Dr。 Ivan Zanoni报道，干扰素λ减少了结肠炎小鼠模型的氧化应激和肠道损伤，并 提示外源性干扰素λ可抑制肠道炎症。重要的是，我们确认了两名没有血缘关系的患者 婴儿发病的炎症性疾病伴有干扰素λ2和干扰素λ3罕见且功能有害的突变。 随着年龄的增长，病情明显改善。我们有初步数据表明干扰素λ2和干扰素λ3可能更重要 在生命的最初几个月，比FNλ1和数据显示更严重的小鼠结肠炎在干扰素λ3-/-小鼠中进行比较 到野生型小鼠。综上所述，我们推测干扰素λS是粘膜的重要调节剂 动态平衡，预防IBD的发展，并保持治疗潜力。目前可用的治疗方法 对于IBD的管理未能治疗大量的患者。这项工作将提供一个更好的理解 干扰素λ在粘膜动态平衡中的作用，并可能为实施新的策略提供基础 通过操纵干扰素λ信号治疗炎症性肠病。解开干扰素λ在维持粘膜动态平衡中的作用 通过以下目的实现：(1)建立干扰素λS的发育性表达 用批量和单细胞RNA测序技术研究不同年龄人群中的细胞因子和受体(2) 用不同小鼠研究干扰素λ信号在体内结肠炎易感性中的作用 结肠炎模型；(3)体外研究患者编码的干扰素λ变异体的功能后果。 T84细胞以及人类对照和患者来源的肠道器官。K08奖在 使Ouahed博士获得的必要的结构化培训能够熟练掌握关键技能： RNAseq分析、小鼠结肠炎模型和免疫分析，以及肠道的产生/操作 有机物和上皮细胞分析，辅以说教课程，帮助她走上独立之路 作为一名成功的内科科学家。 本附录的目的是为一名全职研究助理提供12个月的资金。这 额外的支持对于确保瓦赫德博士在期间和立即继续提高工作效率至关重要 在她即将休完产假之后。