CAREER: Organelle Networks Orient Cilia and First-Generation College Students for Success

该奖项全部或部分根据2021年美国救援计划法案（公法117-2）资助。该项目的目标是调查在细胞表面定位纤毛的机制。 纤毛是进化上古老的毛发状附属物，对细胞运动和感觉至关重要。 了解纤毛是如何定位的是很重要的，因为纤毛的位置影响陆生和水生生物如何获得营养，感知环境和繁殖。 这项研究是基于这样的假设，即相互连接的细胞器网络决定纤毛在哪里形成，纤毛如何感知分子，以及纤毛如何移动流体。 这项研究将由一个主要由本科生科学家组成的团队进行，他们将获得尖端荧光显微镜，图像分析和进化分歧细胞遗传操作的经验。 通过该提案产生的科学数据将用于创建一个跨学科的数学建模课程，用于纤毛内的教学扩散。 除了教学和科学研究，该项目的一个主要目标是为有抱负的第一代大学生形成一个纵向指导计划。 在该项目的第一年，将从当地一所有资格获得联邦大学准备援助（GEAR UP）的低收入高中召集一批9年级学生。 在他们的高中生涯中，这群人将从事细胞生物学研究，以了解当地河流的水污染如何影响纤毛运动。 学生的科学培训将与家庭参与，丰富活动和大学申请援助相结合，以增加这些学生在项目完成时参加STEM本科课程的可能性。 美国救援计划资助这个项目支持这位研究人员在他的职业生涯的关键阶段。纤毛是基于微管的结构，其从跨越单细胞生物（如四膜虫）的细胞表面突出，通过多细胞生物（如人类）。 纤毛的分子结构是高度保守的;在大多数研究的生物体中，纤毛由9个径向对称的双线微管组成，称为轴丝，组装在9个径向对称的三线微管之上，称为基体。 尽管纤毛分子结构具有很强的保守性，但纤毛相对于细胞几何形状的物理位置并不保守。 这个项目利用进化上不同的细胞来测试假设如何膜结合细胞器邻近基体参与纤毛的定位，信号和弯曲特性。 这项研究将调查单细胞原生动物四膜虫和来自小鼠和人类的细胞培养物中膜结合细胞器和基体之间的相互作用。 这种比较方法的目的是确定使用动态活细胞荧光显微镜，Förster共振能量转移（FRET）定量荧光寿命成像显微镜（FLIM），比率钙显微镜，药理学和遗传扰动纤毛定位的保守机制。 总的来说，该项目的结果将揭示影响细胞如何相对于细胞几何形状定位纤毛的基本机制。 这些结果的影响，纤毛是如何组织，以最大限度地提高细胞的运动性在水生环境和信号的转导从胞外space.This奖项反映了NSF的法定使命，并已被认为是值得通过使用该基金会的智力价值和更广泛的影响审查标准进行评估的支持。

This award is funded in whole or in part under the American Rescue Plan Act of 2021 (Public Law 117-2).The goal of this project is to investigate the mechanisms that position cilia across the surface of cells. Cilia are evolutionarily ancient hair-like appendages that are critical for cell movement and sensation. Understanding how cilia are positioned is important because the position of cilia affects how both terrestrial and aquatic organisms acquire nutrients, sense their environment, and reproduce. This research is based on the hypothesis that networks of interconnected organelles determine where cilia form, how cilia sense molecules, and how cilia move fluid. This research will be conducted by a team of primarily undergraduate student scientists, who will gain experience in cutting-edge fluorescence microscopy, image analysis, and genetic manipulation of evolutionarily divergent cells. The scientific data generated through the proposal will be used to create an interdisciplinary mathematical modeling curriculum for teaching diffusion within cilia. Beyond teaching and scientific research, a major goal of this project is to form a longitudinal mentoring program for aspiring first-generation college students. During year 1 of the project, a cohort of 9th grade students will be assembled from a local low-income high school that qualifies for federal college preparation assistance (GEAR UP). Throughout their high school career, this cohort will engage in cell biology research to understand how water pollution in a local river impacts cilia movement. Student scientific training will be complemented with family engagement, enrichment activities and college application assistance to increase the likelihood that these students enroll in STEM undergraduate programs at the completion of the project. American Rescue Plan funding of this project supports this researcher at a critical stage in his career. Cilia are microtubule-based structures that project from the surface of cells that span unicellular organisms, such as Tetrahymena, through multi-cellular organisms, such as humans. The molecular architecture of cilia is deeply conserved; in most organisms examined, cilia are composed of 9 radially symmetric doublet microtubules, called an axoneme, assembled on top of 9 radially symmetric triplet microtubules, called a basal body. Despite the strong conservation of cilia molecular architecture, the physical location of cilia relative to the geometry of the cell is not conserved. This project utilizes evolutionarily divergent cells to test hypotheses for how membrane-bound organelles adjacent to basal bodies participate in the positioning, signaling, and bending properties of cilia. The research will investigate interactions between membrane-bound organelles and basal bodies in both the unicellular protist Tetrahymena and cell cultures derived from mice and humans. This comparative approach aims to identify conserved mechanisms of cilia positioning using dynamic live-cell fluorescence microscopy, Förster Resonance Energy Transfer (FRET) quantified with Fluorescence Lifetime Imaging Microscopy (FLIM), ratiometric calcium microscopy, pharmacology, and genetic perturbations. Overall, the results from this project will reveal fundamental mechanisms that impact how cells position cilia relative to cellular geometry. These results have implications for how cilia are organized to maximize cellular motility in aquatic environments and the transduction of signals from the extracellular space.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.