Performing Department
Plant Biology and Pathology
Non Technical Summary
Trichothecenes areubiquitoustoxic secondary metabolites produced by pathogenic fungi in host plants such as wheat, barley, oats, and maize. Due to their natural occurrence in food and high toxicity, trichothecene contamination of food and livestock feed (>78% toxin present) poses a great threat to food safety and human, animal, and planthealth.However, there is a critical gap in our knowledge about the way the toxin works in living cells leading to toxicity that endangersanimals and humans. Previous studies identified novel mechanisms for trichothecene-induced cell death involving themitochondria, an important component of thecell that functions to help produce energy. These studies have shown that the toxin-induced cell death involved mitochondria and genes involved in lipid metabolisms which mediated sensitivity to these toxins. Therefore, the proposed project will study the role oflipid signaling as a mitochondrial 'surveillance' mechanism induced in response to trichothecene-mediated stress.Applying high-throughput methods in molecular biology and biochemistry, this research will identify lipids altered in response to toxin treatment and determine if lipid signaling is important for trichothecene-mediated mitochondrial fragmentation. It will also measure therecognition and degradation of trichothecene-damaged mitochondria by mitophagy, a selective degradation of damaged mitochondria. The research will identify critical components of the mitochondrial-nuclear signaling pathways using yeast (highly conserved between yeasts and animal cells) and then investigate them in more detail in animal cells. This research will leadto new insights into how these toxins modulate themitochondria and mitochondrial signaling and advance our understanding of mechanisms whereby environmental toxins derail the fate of mitochondria in the affected animals. The knowledge gained here will aid efforts to mitigate the impacts of trichothecene contamination in food and food safety; facilitating the development of safer practice and standards in human and animal consumption of contaminated products.Ultimately, the project will aid in promoting safer food, developing potential applications for animal health improvement, and reducing economic losses due to trichothecene contamination.
Animal Health Component
5%
Research Effort Categories
Basic
100%
Applied
0%
Developmental
0%
Goals / Objectives
There are twoprimary goals, research and professional development, in this project. First major goal is toidentify critical components of mitochondrial signaling in response to trichothecene-mediated oxidative stress. The project aims to discover the role of lipid and lipid signaling in trichothecene toxicity and provide new ways to quantify mitophagy. Accordingly, the project will address the following objectives:1) Determine the role of lipid signaling in trichothecene-induced mitochondrial fragmentation and mitophagy.2) Determine the role of sphingolipid signaling in trichothecene-induced mitophagy.3) Validate trichothecene-induced mitochondria fragmentation in mammalian cell.Second goal is the professional training and career development of the NIFA postdoctoral fellow. Accordingly, the following objectives are:1) Develop technicalexpertise and knowledge in molecular and biotechnology and toxin mechanism studies including imaging and analysis, flow cytometry, LC-MS/MS, transformation technologies, etc.2) Advance teaching competencies by developing a course focused on science related to postdoctoral fellow's expertisesuch as plant biology, pathology,and microbiologyand/or scientific communication skill development such as grantsmanship.3) Obtain research independence through publications and generating new grant proposals.4) Leadership and communication skill development through outreach activities such as presentation of research at scientific meetings, guest lectures, publications (formal and informal), and student mentoring in aspects of scientific research and grant writing.
Project Methods
Research Methods Overview: The research project will be conducted through usage of standardized and novel methods. First, shotgun and targeted lipidomics assays of yeast treated with toxins. Data will be analyzed and confirmed using statistical analysis. Combination of LC-MS/MS and targeted multiple reaction monitoring (MRM)method will be completed at Michigan Regional Comprehensive Metabolomics Research Core to quantify lipid mediators present. Thin-layer chromatography will be used to identify specific lipids identified through LC-MS and MRM. A novel, quantitative assay for mitophagy using fluorescent reporter Rosella fused to mitochondrial targeting sequence will be developed.The developed method will presentimaging; mitochondrial fragmentation and ROS generation will be quantified and measured.Finally, mammalian cells and toxin induced mitochondrial fragmentation in Vero cells will bedetermined.Efforts: The dissemination of the research and professional development will be completed through formal and informal ways. In addition to classroom and laboratory teaching, I will be developing hybridworkshops and/or courses to assist undergraduate and graduate students' interest in research and science communication.I will provide opportunities for students to be engaged in a laboratory research and provide mentoring. Several outreach activities will be focused in encouraging and assisting students in STEM obtain research freedom by obtaining external funding and further developing research related skills.The research briefs will be distributedto relevant agencies and the information will be available and easily accessible through online sites. In addition, when appropriate,I will request relevantagencies to feature the research project in their newsletters and websites. Evaluation:Research and professional development progress will bereviewed and revisited on a regular basis.Newly generated data will be presented during lab meetingsand I will obtain feedback. The progression will be tracked through progress reports. Goals and objectives will be evaluatedfew times a year to modify and adjustresearch plans.Conference papers andmanuscripts will be evaluated by peersand external reviewers.The planning and creation of workshops and/or courses geared towards undergraduate and graduate students in STEM disciplines (research or grant writingfocused content)will be evaluated by faculty members atRutgers. Once approved, the success of the course and/or workshops will be evaluated through surveys anddiscussions.