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.

项目摘要/摘要 锌稳态失调可能导致锌缺乏或锌过量，从而导致多种多样 人类病理。因此，严格对锌运输和存储的调节对于细胞功能至关重要 和人类健康。可以通过多种模型系统对锌生物学进行强大的研究，I 建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 人类细胞培养物，以对锌生物学有详细而有用的理解。我早期的研究 博士后训练发现，冰染色体相关的细胞器，秀丽隐杆线虫中称为肠颗粒 以锌的依赖方式重组；我确定了一个膨胀室，该隔室的体积增加了锌 过度且条件不足。此外，低锌稳态途径似乎在C中是保守的。 秀丽隐杆线和人类，因为在启动子中已经确定了低锌激活增强子元件 在这两个生物体中起作用低锌反应的锌转运蛋白。为基于这些初步结果， 我建议采用一种多学科的方法来了解锌稳态的监管程度低以及如何 溶酶体进行了重组。在AIM 1中，我建议表征依赖锌的扩展室和 秀丽隐杆线虫中肠道肠颗粒的膜结构。在AIM 2中，我将阐明锌的调节 人类溶酶体的贩运和形态重组。在AIM 3中，我将表征 秀丽隐杆线虫和人类细胞中的低锌途径。 该建议将利用我从细胞生物学，化学和 X射线物理学，并扩展它们以完成我的工具包，以探测遗传学和 生物化学。此外，我将作为模型系统扩展到人类细胞。培训也将基于 我在科学倡导方面的丰富经验，使我成为历史上有力的倡导者 被边缘化的团体作为教师。我在K99阶段的培训将整合我的专业知识 指导Kornfeld博士和Diwan博士完成了我为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 成员成为我作为科学家和科学活动家的职业生涯。 我的长期职业目标是领导一支由多样化的学员进行尖端技术来探究关键的团队 锌稳态和贩运的问题。我的记录很强，证明了我作为科学家的能力， 因此，我作为主要研究人员的潜力。我致力于进行领先的科学和 促进机构变革以促进学术界的多样性和包容性。 K99/R00奖将最大化 我通过提供关键的资源和联系来实现目标的机会 否则我的培训缺乏。