Regulation of zinc-dependent lysosome morphological restructuring, zinc trafficking and low zinc homeostasis in C. elegans and human model systems

秀丽隐杆线虫和人类模型系统中锌依赖性溶酶体形态重组、锌运输和低锌稳态的调节

• 批准号: 10674743
• 负责人: Adelita D. Mendoza
• 金额: $ 12.5万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10674743
• 关键词: Academia; Advocacy; Advocate; Affect; Animals; Architecture; Award; Binding; Biochemistry; Biological Models; Biological Process; Biology; Caenorhabditis elegans; Cell Culture Techniques; Cell physiology; Cells; Cellular biology; Chemistry; Cytoplasm; Cytoplasmic Granules; Dietary Zinc; Elements; Enhancers; Faculty; Fluorescence Microscopy; Gene Expression; Genes; Genetic; Genetic Enhancer Element; Genetic Screening; Genetic Techniques; Genetic Transcription; Goals; Health; Homeostasis; Human; Human Pathology; Institution; Intestines; Large Intestine; Lysosomes; Mediating; Medical; Medical Research; Membrane; Mentors; Messenger RNA; Metabolism; Microscopy; Modeling; Molecular Genetics; Monitor; Morphology; Nutrient; Organelles; Organism; Pathology; Pathway interactions; Pattern; Phase; Physics; Preparation; Process; Proteins; Regulation; Regulatory Pathway; Research Personnel; Resources; Roentgen Rays; Science; Scientist; Signal Pathway; Signal Transduction Pathway; Structure; System; Techniques; Testing; Time; Training; Transcriptional Activation; Transmission Electron Microscopy; Vesicle; ZIP protein; Zinc; Zinc deficiency; career; dietary manipulation; diversity and inclusion; experience; experimental study; gene function; genetic manipulation; human disease; human model; improved; innovation; interdisciplinary approach; marginalized population; member; mutant; novel therapeutic intervention; post-doctoral training; promoter; response; superresolution microscopy; trafficking; transcription factor; zinc-binding protein

Project Summary/Abstract Dysregulation of zinc homeostasis can lead to either zinc deficiency or zinc excess, resulting in a variety of human pathologies. Therefore, strict regulation of zinc trafficking and storage are essential for cellular function and human health. Robust studies of zinc biology can be conducted with a variety of model systems, and I propose to combine the genetic and experimental power of Caenorhabditis elegans with the medical relevance of human cell culture to develop a detailed and useful understanding of zinc biology. Studies from my early postdoctoral training have uncovered that lysosome-related organelles, called gut granules in C. elegans, are restructured in a zinc dependent manner; I identified an expansion compartment that increases in volume in zinc excess and deficient conditions. In addition, the low zinc homeostasis pathway appears to be conserved in C. elegans and humans, since the Low Zinc Activation enhancer element has been identified in the promoters of zinc transporters that function the low zinc response in both organisms. To build upon these preliminary results, I propose to take a multidisciplinary approach to understand how low zinc homeostasis is regulated and how lysosomes are restructured. In Aim 1, I propose to characterize the zinc-dependent expansion compartment and the membrane architecture of intestinal gut granules in C. elegans. In Aim 2, I will elucidate the regulation of zinc trafficking and morphological restructuring of human lysosomes. In Aim 3, I will characterize the regulation of the low zinc pathway in C. elegans and human cells. This proposal will capitalize on my experience with interdisciplinary techniques from cell biology, chemistry, and X-Ray physics, and expand upon them to complete my toolkit for probing zinc in biology with genetics and biochemistry. Furthermore, I will expand into human cells as a model system. The training will also build upon my extensive experience in science advocacy and equip me to be a powerful advocate for historically marginalized groups as a faculty member. My training in the K99 phase will integrate the expertise from my mentor Dr. Kornfeld and co-mentor Dr. Diwan to complete my preparation for the R00 independent phase. Training during the K99 phase will integrate the experience from my mentors, collaborators, and advisory board members to springboard my career as a scientist and science activist. My long-term career goal is to lead a team of diverse trainees performing cutting edge techniques to probe critical questions in zinc homeostasis and trafficking. I have a strong record demonstrating my abilities as a scientist, and therefore my potential as a primary investigator. I am committed to conducting leading edge science AND promoting institutional change to promote diversity and inclusion in academia. The K99/R00 award will maximize my chances of being able to achieve my goals by providing critical resources and connections that would be otherwise be lacking in my training.

项目概要/摘要 锌稳态失调可导致锌缺乏或锌过量，从而导致多种疾病 人类病理学。因此，严格调节锌的运输和储存对于细胞功能至关重要 和人类健康。锌生物学的稳健研究可以使用各种模型系统进行，并且我 提议将秀丽隐杆线虫的遗传和实验能力与医学相关性结合起来 人类细胞培养的研究，以加深对锌生物学的详细和有用的了解。从我早年开始学习 博士后培训发现，线虫中与溶酶体相关的细胞器（称为肠道颗粒） 以锌依赖方式重组；我发现了一个可以增加锌体积的膨胀室 过剩和不足的条件。此外，低锌稳态途径似乎在 C. 线虫和人类，因为低锌激活增强子元件已在 锌转运蛋白在两种生物体中发挥低锌反应的作用。为了以这些初步结果为基础， 我建议采取多学科方法来了解低锌稳态是如何调节的以及如何调节 溶酶体被重组。在目标 1 中，我建议描述锌依赖性膨胀室的特征，并 线虫肠道颗粒的膜结构。在目标 2 中，我将阐明锌的调节 人类溶酶体的贩运和形态重组。在目标 3 中，我将描述 线虫和人类细胞中的低锌途径。 该提案将利用我在细胞生物学、化学和跨学科技术方面的经验。 X 射线物理学，并对其进行扩展以完成我的工具包，用于通过遗传学和 生物化学。此外，我将扩展到人类细胞作为模型系统。该培训还将建立在 我在科学倡导方面的丰富经验使我能够成为历史上强有力的倡导者 边缘群体作为教员。我在 K99 阶段的培训将整合我的专业知识 导师 Kornfeld 博士和共同导师 Diwan 博士完成了我 R00 独立阶段的准备工作。 K99阶段的培训将整合我的导师、合作者和顾问委员会的经验 成员们为我作为科学家和科学活动家的职业生涯提供了跳板。 我的长期职业目标是领导一支由多元化学员组成的团队，运用尖端技术来探索关键问题 锌稳态和贩运的问题。我有良好的记录证明我作为科学家的能力， 因此我有成为一名主要研究者的潜力。我致力于开展前沿科学并且 促进制度变革，以促进学术界的多样性和包容性。 K99/R00奖项将最大化 我通过提供关键资源和联系来实现我的目标的机会 否则我的训练就会有所欠缺。