BRIGE: A reductionist approach to enterovirus disinfection

PI：Wigginton Proposal number：1228076这项布里奇提案描述了一项致力于对肠道病毒颗粒在消毒过程中发生的化学反应进行简化论分析的计划。该项目是在马里兰大学建立一个研究团队的长期目标的第一步，该团队致力于使用最先进的分析技术来检测和表征病原微生物。为了了解在病毒复合体中发生的反应，我们必须首先了解蛋白质和基因组亚成分的反应性。尽管蛋白质、核酸单体和常见消毒剂之间的反应已有了很好的描述，但更高级别的组织对单体反应活性的作用却知之甚少。有人建议开发一种自下而上的方法来描述脊髓灰质炎病毒颗粒与游离氯的反应性。建立了一种基质辅助激光解吸电离电离(MALDI-MS)定量技术，用于测定氯与结构日益复杂的生物聚合物(即多肽、RNA低聚物、蛋白质、基因组、衣壳、感染性病毒)之间的反应动力学和产物。在这样做的过程中，将解决有关病毒分子结构在其消毒敏感性中所起作用的基本问题。将比较三种类似脊髓灰质炎病毒株的反应性和灭活情况；这将阐明病毒序列的微小变化如何影响其基本反应性和消毒动力学。这项提议的扩大参与方面与研究相结合--它强调留住工程学科的女本科生。具体地说，本科生的研究项目将集中在简单的生物聚合物反应性实验上。本科生研究员将与马里兰大学工程女性研究员计划合作选择和指导。这项提议的扩大参与方面还包括与UMD新的NSF STEP计划的计划活动，该计划名为成功的工程教育和发展支持(SEED)。有了种子，学生们将接触到当前工程学中的病原体研究课题(例如，生物传感器、发展中国家的努力等)。通过午餐时间的讲座和实验室参观。拟议工作的智力价值：这项研究计划将极大地提高对病毒粒子在用次氯酸消毒时发生的反应的理解。到目前为止，科学家们仍然缺乏对病毒灭活的机械性描述，这项工作将作为对导致灭活和不导致灭活的修饰的全面检查。这项工作将带来一系列科学进展，包括开发用于研究病毒蛋白质和基因组的定量MALDI-MS技术，对蛋白质和基因组组织在其对氧化剂的敏感性中所起作用的新视角，对RNA氧化反应的深入评估，以及预测新出现的病毒株的消毒动力学将如何不同于基于核酸和氨基酸突变的常见病毒株的消毒动力学的工具。拟议工作的广泛影响：本文描述的研究工作在水卫生领域具有广泛的潜在影响。新出现的病毒不断给环境工程师和公共卫生科学家带来新的挑战。更好地理解变异对病毒“耐受性”的影响，将有助于开发用于消毒过程设计的敏感性预测工具。此外，对病原体消毒的更好的理解可以导致开发有效的低成本、低能量的消毒策略。

PI: WiggintonProposal Number: 1228076This BRIGE proposal describes a plan dedicated to a reductionist analysis of the chemical reactions that take place in an enterovirus particle during disinfection. The project represents the first step of a long-term goal to develop a research team at the University of Maryland dedicated to employing state-of the art analytical techniques to detect and characterize pathogenic microorganisms. In order to understand the reactions that take place in a virus complex, one must first understand the reactivity of the protein and genome subcomponents. Although the reactions between protein and nucleic acid monomers and common disinfectants are well characterized, the role that higher order organization has on monomer reactivity is poorly understood. It is proposed that a bottom-up approach to describe a poliovirus particle's reactivity with free chlorine be developed. A quantitative Matrix Assisted Laser Desorption Ionization mass spectrometry (MALDI-MS) technique will be developed to determine the kinetics and products of reactions between chlorine and biopolymers with increasing structure complexity (i.e., peptides, RNA oligomers, proteins, genomes, capsids, infective viruses). In doing so, fundamental questions about the role of virus molecular structure in its disinfection susceptibility will be addressed. The reactivity and inactivation of three similar poliovirus strains will be compared; this will elucidate how slight changes in a virus sequence influence its fundamental reactivity and disinfection kinetics. The broadening participation aspect of this proposal is integrated with the research-it emphasizes retaining female undergraduate students in engineering disciplines. Specifically, an undergraduate research project will focus on the simple biopolymer reactivity experiments. The undergraduate researcher will be selected and mentored in collaboration with the University of Maryland Women in Engineering Fellows Program. Also included in the broadening participation aspect of this proposal are planned activities with the new NSF STEP program at UMD titled Successful Engineering Education and Development Support (SEEDS). With SEEDS, students will be exposed to current pathogen research topics in engineering (e.g., biosensors, efforts in the developing world, etc.) through lunchtime lectures and lab tours. Intellectual Merit of Proposed Work: This research plan will substantially improve the understanding of the reactions that take place in a virus particle as it is disinfected with hypochlorous acid. To date, scientists continue to lack a mechanistic description of virus inactivation and this work will serve as a holistic examination of the modifications that do and do not contribute to inactivation. The work will result in a number of scientific advancements including the development of a quantitative MALDI-MS technique for studying virus proteins and genomes, a new perspective on the role of protein and genome organization in their susceptibility to oxidants, an in-depth evaluation of RNA oxidation reactions, and tools to predict how the disinfection kinetics of emerging virus strains will differ from the disinfection kinetics of familiar virus strains based on nucleic acid and amino acid mutations.Broader Impacts of Proposed Work: The research effort described herein has wide reaching potential impacts in the field of water sanitation. Emerging viruses constantly create new challenges for environmental engineers and public health scientists. A better comprehension of the influence that mutations have in virus "hardiness" will allow for the development of susceptibility prediction tools for use in disinfection process design. Furthermore, an improved understanding on pathogen disinfection could lead to the development of effective low-cost, low-energy, disinfecting strategies.