Supply and Demand of Proteins During Neuronal Growth and Extension

本项目研究轴突生长过程中神经元蛋白的产生、传递和需求之间的基本关系。要验证的整体假设是，轴突运输和局部蛋白质生产是神经元生长所需资源的主要供应商，并在一定程度上受到神经元局部机械环境的调节。这一假设将通过多学科的方法来解决，包括分子和细胞生物学，高分辨率成像和图像处理，以及细胞生物力学。可以选择任意数量的候选蛋白来研究神经元的供求关系。然而，在这个项目中要考虑的具体货物是那些对生长、运输和蛋白质合成至关重要的：线粒体，负责满足神经元对atp依赖的能量需求，液泡，负责提供扩大神经元表面积所需的质膜；核糖体，负责蛋白质合成；和mRNA，为蛋白质合成提供模板转录物。提出的研究的几个方面是变革性的，包括开发新的工程方法来检查和量化神经生物学过程，以及使用系统方法来理解神经元的亚细胞功能。从长远来看，这一基础科学项目为与轴突运输缺陷相关的神经退行性疾病的进展提供了基础，如阿尔茨海默病和卢·格里克病（ALS）。此外，这种理解神经元内生理过程的生物工程方法将促进一种有价值的数据驱动方法来建模和操纵神经元货物供需的动态，以及细胞资源的分配。最后，考虑到细胞系统中运输过程的普遍性，预计从该项目中确定的一般原则将与非神经元细胞的功能和功能障碍有关。除了上述强调的生物医学和基础科学影响外，拟议的计划还影响教育、研究培训和社区外展。在课堂上，研究成果将整合到两门基于细胞生理学的生物工程课程中，这两门课程的课程材料都大量借鉴了当前的研究方法。鉴于拟议研究的多学科性质，将为具有工程和生物科学背景的本科生和研究生提供培训机会。到目前为止，来自不同背景的学生已经成功地招募到PI的实验室，并且基于强调开放，协作的工作环境，预计这一成功将继续下去。这种跨学科、以团队为基础的方法对于培养新一代生物医学科学家和生物工程师至关重要。此外，将启动或继续参与各种外展计划。一个新的神经生物工程研究项目（NBRP）将与埃莉诺·罗斯福高中的QUEST项目相结合，该高中的招生由理工科学生人数不足的学生组成。该计划的部分内容已经开始实施，该计划向初高中学生提供生物工程方面的职业选择，向他们介绍研究方法和科学写作，并为他们提供使用最先进实验室设施的机会。积极参与分子和细胞生物工程本科生研究经验（REU）计划也将继续。优先录取那些在本国机构获得研究机会有限的学生。PI的愿景与Fischell生物工程系、Clark工程学院和马里兰大学的愿景是一致的；最终的目标是鼓励和促进K-12、本科生和研究生参与多学科生物医学研究，努力吸引最优秀的学生进入这个令人兴奋的领域。

0932590ShahINTELLECTUAL MERITThis project addresses the fundamental relationship between the production, delivery, and demand of neuronal proteins during axonal growth. The global hypothesis to be tested is that axonal transport and local protein production are primary suppliers for the resources needed for neuronal growth, and are regulated in part by the local mechanical environment of the neuron. This hypothesis will be addressed by using a multidisciplinary approach involving molecular and cellular biology, high-resolution imaging and image processing, and cell biomechanics.Any number of candidate proteins may be selected to study neuronal supply and demand. However, the specific cargoes to be considered in this project are those essential for growth, transport, and protein synthesis: mitochondria, responsible for fulfilling ATP-dependent energetic requirements for the neuron, vacuoles, responsible for supplying plasma membrane required for expansion of neuronal surface area; ribosomes, responsible for protein synthesis; and mRNA, which provides the template transcript for protein synthesis.Several aspects of the proposed research are transformative, including the development of novel engineering approaches to examine and quantify neurobiological processes, and the use of a systems approach to understand the subcellular function of neurons. In the long term, this basic science project has implications for providing a basis for the progression of neurodegenerative diseases linked to defects in axonal transport, such as Alzheimer's and Lou Gehrig's Disease (ALS). Additionally, this bioengineering approach to understanding physiological processes within the neuron will facilitate a valuable data-driven approach to modeling and manipulating the dynamics of neuronal cargo supply and demand, and the allocation of cellular resources. Finally, given the ubiquity of transport processes in cellular systems, it is expected that general principles identified from this project will be relevant to the function and dysfunction of non-neuronal cells. BROADER IMPACTS In addition to the biomedical and basic scientific impacts highlighted above, the proposed plan also impacts education, research training, and community outreach. In the classroom, research results will be integrated into two bioengineering courses based on cell physiology, both of which draw heavily on current research approaches for their course material. Given the multi-disciplinary nature of the proposed research, training opportunities for undergraduate and graduate students from backgrounds in both engineering and biological sciences will be provided. Thus far, students from multiple backgrounds have successfully been recruited to the laboratory of the PI, and the continuation of this success is anticipated based on the emphasis on an open, collaborative work environment. This cross-disciplinary, team-based approach is critical for training a new generation of biomedical scientists and bioengineers. Additionally, participation in a variety of outreach programs will be initiated or continued. A new Neuro-Bioengineering Research Program (NBRP) will be created, in conjunction with the QUEST program at Eleanor Roosevelt High School, whose enrollment consists of student populations under-represented in science and engineering. Implementation has already been started for portions of this program, which exposes middle and high school students to career options in bioengineering, introduces them to research methodology and scientific writing, and provides them with access to state-of-the-art laboratory facilities. Active participation in the Molecular and Cellular Bioengineering Research Experiences for Undergraduates (REU) Program will also be continued. It is a priority to admit students to this program who have limited access to research opportunities at their home institutions. The vision of the PI is consistent with that of the Fischell Department of Bioengineering, the Clark School of Engineering, and the University of Maryland; ultimately, the aim is to encourage and facilitate the participation of K-12, undergraduate, and graduate students in multidisciplinary biomedical research, in an effort to attract the finest students to this exciting field.