Gene-environment interactions in necrotizing enterocolitis: impact of SIGIRR mutation and gut microbiota on intestinal TLR hyperactivity

项目摘要/摘要 这份NIH K08提案描述了一名内科科学家候选人的五年培训和研究计划 公司的长期目标是成为新生儿医学领域的独立研究者。朝向这个方向 最后，拟议的研究旨在确定坏死性小肠结肠炎(NEC-a)的发病机制。 早产儿的破坏性炎症性肠病，其主要特征是过量 激活Toll样受体(TLR)家族的天然免疫受体。固有遗传易感性 肠道微生物区系环境是与NEC发病密切相关的两个因素。这些 评估因素主要是相互隔离的--这是一个重要的限制，不 复制NEC的复杂性。 最近，我们实验室培育出了一种新的转基因小鼠，它重现了在婴儿身上发现的一种突变。 与NEC合作。该突变是单一免疫球蛋白白介素1相关受体(SIGIRR)的停止突变， 一种正常作用于抑制肠道TLR信号的基因。产生的SIGIRR转基因小鼠(SIGIRRTG) 在基线条件下表现出肠道TLR活性增加的表型。在这项研究中，候选人提出了 用这一新的小鼠品系作为NEC的遗传模型，以确定遗传学和 肠道微生物群，因为它与肠道TLR活性有关。目标1将确定SIGIRR突变对肠道的影响 微生物区系特征。目的#2将确定SIGIRRTG小鼠肠道TLR活性增加是否 由肠道微生物区系驱动。目的#3将确定肠道微生物群改变在逆转遗传方面的作用 SIGIRRTG小鼠对肠道炎症的易感性。完成这些目标将提供新的 深入了解基因和肠道微生物群如何相互作用以影响发育中肠道中的TLR动态平衡。 所获得的新知识将代表着对复杂的发病机制的理解的重大进步。 NEC，可用于为NEC的个性化、基于微生物组的疗法的开发提供信息。 候选人坚定地致力于基础和翻译NEC研究的职业生涯，并坚定地 在他的职业生涯和研究目标上得到了他的导师和他在堪萨斯儿童慈善机构的支持 城市。他目前担任儿科学助理教授，有75%的保护时间 研究、独立实验室和办公空间，以及启动资金。目前的提案包括一项 全面的指导和教学计划，以提高候选人在TLR生物学方面的技能和知识， 微生物发病机制、细胞和分子生物学、微生物组测序、生物信息学和研究 伦理道德。完成这项全面的培训计划将为应聘者提供技能和经验 有必要制定独立的研究计划并获得R01资金。

PROJECT SUMMARY/ABSTRACT This NIH K08 proposal describes a five-year training and research plan for the candidate, a physician-scientist with a long-term goal of becoming an independent investigator in the field of neonatal medicine. Toward this end, the proposed research is directed at defining the pathogenesis of necrotizing enterocolitis (NEC) – a devastating inflammatory bowel disease of preterm infants that is characterized mainly by the excessive activation of the Toll-Like Receptor (TLR) family of innate immune receptors. Inherent genetic susceptibility and the intestinal microbiota environment are two factors strongly implicated in NEC pathogenesis. These factors have primarily been evaluated in isolation from each other – an important limitation that does not replicate the complex nature of NEC. Recently, our laboratory generated novel transgenic mice that recapitulate a mutation discovered in infants with NEC. This mutation is a stop mutation of Single-Immunoglobulin Interleukin-1 Related Receptor (SIGIRR), a gene that normally functions to inhibit gut TLR signaling. The generated SIGIRR transgenic mice (SIGIRRTG) exhibit a phenotype of increased gut TLR activity at baseline conditions. In this study, the candidate proposes to use this novel mouse line as a genetic model of NEC to determine the interaction between genetics and the gut microbiome as it relates to gut TLR activity. Aim #1 will determine the impact of SIGIRR mutation on gut microbiota signatures. Aim #2 will determine whether increased intestinal TLR activity in SIGIRRTG mice is driven by gut microbiota. Aim #3 will determine the effect of gut microbiome alteration in reversing genetic predisposition of SIGIRRTG mice towards intestinal inflammation. Completion of these aims will provide novel insights into how genes and the gut microbiome interact to influence TLR homeostasis in the developing gut. New knowledge gained will represent a significant advancement in understanding the complex pathogenesis of NEC that can be used to inform the development of personalized, microbiome-based therapies for NEC. The candidate is firmly committed to a career in basic and translational NEC research and is strongly supported in his career and research goals by his mentors and his department at Children’s Mercy Kansas City. He currently holds a position as an Assistant Professor of Pediatrics with 75% protected time for research, independent laboratory and office space, and start-up funding. The current proposal includes a comprehensive mentorship and didactic plan to advance the candidate’s skills and knowledge in TLR biology, microbial pathogenesis, cell and molecular biology, microbiome sequencing, bioinformatics, and research ethics. Completion of this comprehensive training plan will provide the candidate with the skills and experience necessary to develop an independent research program and obtain R01 funding.