CAREER: Using C. elegans to Understand how a Fundamental Cellular Stress Response is Integrated into a Tissue System at the Interface with the Environment and to Improve Education

对于细胞外基质屏障和诱导型细胞保护转录因子如何共同检测环境胁迫和促进体内平衡，我们知之甚少。秀丽隐杆线虫有一个单一的抗氧化/解毒反应转录因子SKN-1。该奖项的主要研究者令人惊讶地发现，角质层的渗透和遗传扭曲激活了几个依赖SKN-1的细胞保护基因，SKN-1强烈影响角质层结构和角质层重塑基因的表达。本次CAREER奖研究的核心假设是，SKN-1被角质层相关的环境压力传感器激活，反过来，SKN-1指导或修改角质层在压力或衰老过程中的合成或重塑。线虫每年损害价值800亿美元的农作物，给畜牧业造成负担，并感染全世界三分之一的人类。因此，本项目是了解SKN-1与线虫角质层如何相互作用影响对农药、抗寄生虫药物的抗性以及动植物宿主免疫反应的重要起点。一个新的高级研究课程将培养一代分子遗传学研究的初级科学家，并促进七个秀丽隐杆线虫实验室之间的合作，研究广泛的主题。额外的外展活动将直接改善高中学生和教师的STEM培训和读写能力，其中许多人主要来自服务不足的社区。SKN-1及其同系物是细胞保护基因的主要调控因子，在整个动物王国中促进抗逆性和长寿。该研究的目的是利用分子和遗传学方法在秀丽隐杆线虫中：1)识别从角质层到SKN-1的渗透信号；2)研究SKN-1是否、为什么以及如何改变角质层；3)开发秀丽隐杆线虫学习模块和教师研讨会；4)为七个实验室的初级科学家教授高级秀丽隐杆线虫研究课程。生化分析、RNA干扰方法、转录谱分析、组织学和超微结构分析以及遗传学研究将实现这些目标。渗透压是最广泛共享和最古老的体内平衡形式之一。该项目将为渗透传感和信号转导提供新的遗传和分子见解。秀丽隐杆线虫是一种强大的研究模式（例如，简单和廉价，但允许复杂的遗传和分子方法），出于许多相同的原因，它在教育方面具有巨大的未被满足的潜力。该项目将与一个大学预科培训项目和高中教师合作，开发和实施利用RNA干扰和绿色荧光蛋白技术的学习模块和培训讲习班，以激发人们对基因调控的兴趣，并展示基础研究的广泛应用。这些研究的结果将发表在同行评议的期刊上，并在区域和国家科学会议上提出。

Very little is known about how extracellular matrix barriers and inducible cytoprotective transcription factors function together to detect environmental stress and promote homeostasis. The nematode Caenorhabditis elegans has a single antioxidant/detoxification response transcription factor named SKN-1. The principle investigator of this award has made the surprising discovery that osmotic and genetic distortion of the cuticle activates several SKN-1- dependent cytoprotective genes and that SKN-1 strongly influences cuticle structure and expression of cuticle remodeling genes. The central hypothesis for the research in this CAREER award is that SKN-1 is activated by a cuticle associated environmental stress sensor and that SKN-1, in turn, directs or modifies cuticle synthesis or remodeling during stress or aging. Nematode parasites damage 80 billion dollars worth of crops each year, burden animal husbandry, and infect 1/3 of humans world-wide. Therefore, this project is an important starting point for a basic understanding of how SKN-1 and the nematode cuticle interact to influence resistance to pesticides, anti-parasitic drugs, and plant and animal host-immune responses. A new advanced research course will train a generation of beginning scientists on molecular genetics research and foster collaboration among seven C. elegans laboratories studying a broad range of topics. Additional outreach activities will directly improve STEM training and literacy for high school students and teachers, many from predominantly underserved communities. SKN-1 and its homologs are master regulators of cytoprotective genes that promote stress resistance and longevity throughout the animal kingdom. The study objectives are to leverage molecular and genetic approaches in C. elegans to: 1) identify osmotic signaling from the cuticle to SKN-1; 2) investigate whether, why, and how SKN-1 modifies the cuticle; 3) develop C. elegans learning modules and a teacher workshop; and 4) teach an advanced C. elegans research course for beginning scientists from seven labs. Biochemical assays, RNA interference approaches, transcript profiling, histologic and ultrastructure analyses, and genetic studies will be performed to achieve these objectives. Osmolarity is one of the most broadly shared and ancient forms of homeostasis. This project will provide novel genetic and molecular insights into osmo-sensing and signal transduction. For many of the same reasons that it is a powerful research model (e.g., simple and inexpensive, but permits sophisticated genetic and molecular approaches), C. elegans has tremendous unmet potential in education. The PI will collaborate with an institutional pre-collegiate training program and high school teachers to develop and implement learning modules and training workshops that utilize RNA interference and green fluorescent protein technologies to stimulate interest in gene regulation and demonstrate broad application of basic research. Results from the studies will be published in peer-reviewed journals and presented at regional and national scientific meetings.