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

项目总结/摘要 这份NIH K 08提案描述了一个为期五年的培训和研究计划，为候选人，一个医生，科学家 长期目标是成为新生儿医学领域的独立研究者。朝着这个 最后，本研究旨在明确坏死性小肠结肠炎（NEC）的发病机制， 早产儿的毁灭性炎症性肠病，其主要特征是 先天免疫受体的Toll样受体（TLR）家族的激活。遗传易感性 和肠道微生物群环境是与NEC发病机制密切相关的两个因素。这些 这些因素主要是在相互孤立的情况下进行评估的，这是一个重要的限制， 复制NEC的复杂性。 最近，我们的实验室培育了一种新型转基因小鼠， 关于NEC该突变是单免疫球蛋白白细胞介素-1相关受体（SIGIRR）的终止突变， 一种通常抑制肠道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TLR活性增加的表型。在这项研究中，候选人提出 使用这种新的小鼠品系作为NEC的遗传模型，以确定遗传学和基因之间的相互作用。 肠道微生物组，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 微生物群特征目的#2将确定SIGIRRTG小鼠中肠TLR活性的增加是否是 由肠道微生物群驱动目的#3将确定肠道微生物组改变在逆转遗传性疾病中的作用。 SIGIRRTG小鼠对肠道炎症的易感性。这些目标的实现将提供新的 深入了解基因和肠道微生物组如何相互作用，以影响发育中肠道的TLR稳态。 获得的新知识将代表着在理解复杂的发病机制方面的重大进展， NEC，可用于为NEC的个性化，基于微生物组的治疗提供信息。 候选人坚定地致力于基础和转化NEC研究的职业生涯， 在他的职业生涯和研究目标的支持下，他的导师和他的部门在儿童慈善堪萨斯 市他目前担任儿科助理教授，有75%的时间受到保护， 研究、独立的实验室和办公空间以及启动资金。目前的提案包括一个 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， 微生物发病机理，细胞和分子生物学，微生物组测序，生物信息学和研究 伦理完成这一全面的培训计划将提供候选人的技能和经验 需要开发一个独立的研究计划，并获得R 01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.