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

项目总结/摘要 锌稳态失调可导致锌缺乏或锌过量，导致多种 人类病理学因此，严格调节锌的运输和储存对细胞功能至关重要 和人类健康。锌生物学的稳健研究可以用各种模型系统进行， 建议将秀丽隐杆线虫的遗传和实验能力与医学相关性联合收割机结合起来 人类细胞培养，以发展锌生物学的详细和有用的理解。我早期的研究 博士后研究发现，C.优雅群岛 以锌依赖的方式重组;我确定了一个膨胀室，在锌的体积增加 过剩和不足的条件。此外，低锌稳态途径似乎是保守的，在C。 因为低锌激活增强子元件已经在线虫和人类的启动子中被鉴定， 在两种生物体中起低锌反应作用的锌转运蛋白。在这些初步结果的基础上， 我建议采取多学科的方法来了解低锌稳态是如何调节的， 溶酶体被重构。在目标1中，我建议表征锌依赖性膨胀室， C.肠颗粒的膜构筑。优雅在目标2中，我将阐明锌的调节 人溶酶体的运输和形态学重构。在目标3中，我将描述 C.低锌途径线虫和人类细胞。 这个建议将利用我的经验与跨学科的技术，从细胞生物学，化学， X射线物理学，并扩大他们完成我的工具包探测锌在生物学与遗传学和 生物化学此外，我将扩展到人类细胞作为模型系统。培训还将建立在 我在科学宣传方面的丰富经验使我能够成为历史上一个强有力的倡导者。 作为一名教员的边缘群体。我在K99阶段的培训将整合我的专业知识， 指导Kornfeld博士和共同指导Diwan博士完成R 00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 作为一名科学家和科学活动家的跳板。 我的长期职业目标是带领一个由不同学员组成的团队，运用尖端技术， 锌稳态和运输的问题。作为一名科学家，我有很强的能力， 因此我有可能成为一名主要调查员我致力于进行前沿科学， 促进机构变革，以促进学术界的多样性和包容性。K99/R 00奖项将最大限度地 通过提供关键的资源和关系，我有机会实现我的目标， 否则我的训练就不够了

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.