Collaborative Research: RUI: Comparative analysis of endocytic trafficking during cell division

本研究项目探讨细胞分裂和细胞交换以协调其行为的信号之间的相互作用。这是一个关键的研究领域，因为细胞行为的信号依赖协调对所有生命形式的生长都是必不可少的。信号传导在干细胞分裂中也起着至关重要的作用，确保有足够数量的干细胞参与组织更新和修复。此外，信号传导错误是癌症的主要原因。由于癌症也需要不受控制的细胞分裂，因此了解分裂如何加剧信号错误尤为重要。为了探索这个根本重要的问题，这个合作研究项目包括三个机构，普林斯顿大学，斯沃斯莫尔学院和科尔比学院。联合的专业知识将使利用一系列生物体以及尖端的遗传操作和成像技术对分裂细胞中的信号传导进行全面的研究。此外，此次合作还将开展一项雄心勃勃的扩展计划，其中包括：1)为不同的本科生群体提供丰富的研究经验；2)在一个服务不足的学区为小学生开设一门以癌症生物学为重点的课程，作为斯沃斯莫尔学院已建立的儿童科学暑期项目的一部分；3)科尔比大学暑期学者项目的整合；这两所大学的重点是为纽约梅隆大学和其他多元化的学生做好准备，帮助他们顺利完成大学过渡，并为普林斯顿大学的博士后提供在科尔比教学中心接受教师培训的机会，这些博士后将在科尔比一月学期的项目中授课。受体和相关膜蛋白的细胞运输可以深刻地影响细胞信号传导。由于长期以来的假设，膜运输在有丝分裂期间被关闭，目前的研究尚未解决有关分裂细胞运输的性质和调节的基本问题。特别是，有丝分裂激酶对内吞运输的贡献仍然缺乏特征。此外，特定内吞途径的作用尚未明确。该项目旨在通过两种实验系统的合作来填补这些关键的空白，在这两种实验系统中，先前的研究已经证明了有丝分裂运输的重要作用：哺乳动物表皮细胞的极化和基础索动物（Ciona ninteinalis）心脏祖细胞的诱导。它将调查两个假设；1)细胞周期蛋白依赖性激酶1 （CDK1）以货物特异性的方式抑制溶酶体降解。2) CDK1和极光激酶（AurK）在有丝分裂过程中广泛抑制循环。这些假设将通过完善的方法进行检验，这些方法会破坏特定的有丝分裂激酶和内吞途径，并采用高分辨率实时成像分析来可视化两种模型系统中受体运输的影响。通过对两种高度分化的细胞类型中的一系列货物蛋白进行这些研究，这些发现将为有丝分裂运输对以下方面的影响提供见解：1)发育胚胎的命运决定；2)分裂上皮片极性的重建；3)干细胞或分化谱系的信号依赖行为。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

This research project explores the interplay between cell division and the signals that cells exchange to coordinate their behavior. This is a critical area of investigation because signal-dependent coordination of cell behavior is essential for the growth of all life forms. Signaling also plays a crucial role in dividing stem cells, ensuring that the proper number of stem cells are available to participate in tissue renewal and repair. Additionally, errors in signaling are the primary cause of cancer. Because cancer also entails unregulated cell division, it is particularly important to understand how division may exacerbate signaling errors. To explore this fundamentally important question, this collaborative research project encompasses three institutions, Princeton University, Swarthmore College and Colby College. The combined expertise will enable comprehensive investigations of signaling in dividing cells using an array of organisms, along with cutting edge genetic manipulation and imaging technologies. Additionally, this collaboration will pursue an ambitious outreach program which will include – 1) providing enriching research experiences for a diverse group of undergraduates, 2) development of a class focused on cancer biology for elementary school students in an underserved school district as part of the established Science for Kids summer program at Swarthmore College, 3) integration between Summer Scholars Programs at Colby, Princeton and Swarthmore which focus on preparing URM and other diverse students for a successful college transition and 4) teacher-training opportunities at the Colby Center for Teaching and Learning for Princeton post-docs who will be teaching courses in the Colby January Term program. Cellular trafficking of receptors and associated membrane proteins can profoundly impact cell signaling. Due to the long-held assumption that membrane trafficking is shut down during mitosis, current research has not addressed fundamental questions regarding the nature and regulation of trafficking in dividing cells. In particular, the contributions of mitotic kinases to endocytic trafficking remain poorly characterized. Additionally, the roles of specific endocytic pathways have not been delineated. This project aims to fill these critical gaps through a collaborative effort incorporating two experimental systems in which prior research has demonstrated an essential role for mitotic trafficking: polarization of mammalian epidermal cells and induction of heart progenitor cells in the basal chordate, Ciona intestinalis. It will investigate two hypotheses; 1) Cyclin dependent kinase 1 (CDK1) suppresses lysosomal degradation in a cargo-specific manner. 2) CDK1 and Aurora Kinase (AurK) suppress recycling broadly during mitosis. These hypotheses will be examined with well-established methods that disrupt specific mitotic kinases and endocytic pathways and employ high-resolution live-imaging assays to visualize the impact on receptor trafficking in both model systems. By conducting these studies on a range of cargo proteins in two highly divergent cell types, these findings will provide insights into the impact of mitotic trafficking on – 1) fate decisions in developing embryos 2) re-establishment of polarity in dividing epithelial sheets and 3) signal dependent behaviors of stem cells or differentiated lineages.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.