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

这项研究项目探索了细胞分裂和细胞为协调其行为而交换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于所有生命形式的生长都是必不可少的。信号在干细胞分裂中也起着至关重要的作用，确保有适当数量的干细胞参与组织更新和修复。此外，信号错误是癌症的主要原因。由于癌症还涉及不受调控的细胞分裂，因此了解分裂如何加剧信号错误尤为重要。为了探索这个根本上重要的问题，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。这些结合的专业知识将使人们能够利用一系列生物，以及尖端的基因操作和成像技术，对细胞分裂中的信号进行全面研究。此外，这项合作还将实施一项雄心勃勃的推广计划，包括：1)为不同的本科生群体提供丰富的研究经验；2)作为斯沃斯莫尔学院既定的儿童科学暑期计划的一部分，在服务不足的学区为小学生开设一个专注于癌症生物学的班级；3)整合科尔比、普林斯顿和斯沃斯莫尔的暑期奖学金计划，专注于为URM和其他不同的学生做好成功的大学过渡准备；4)在科尔比教学中心为普林斯顿大学博士后提供教师培训机会，他们将在科尔比一月份的学期计划中教授课程。受体和相关膜蛋白的细胞运输可以深刻地影响细胞信号转导。由于长期以来的假设，膜运输在有丝分裂过程中被关闭，目前的研究还没有解决有关分裂细胞贩运的性质和调控的基本问题。具体地说，有丝分裂酶对胞内转运的贡献仍然没有得到充分的描述。此外，特定的内吞途径的作用还没有被描绘出来。该项目旨在通过两个实验系统的合作来填补这些关键空白，在这两个实验系统中，先前的研究已经证明了有丝分裂运输的关键作用：哺乳动物表皮细胞的极化和在基础脊索中诱导心脏祖细胞。它将研究两个假说：1)细胞周期蛋白依赖性激酶1(CDK1)以一种货物特异性的方式抑制溶酶体的降解。2)CDK1和Aurora Kinase(AurK)在有丝分裂过程中广泛抑制循环。这些假说将用成熟的方法来检验，这些方法扰乱特定的有丝分裂酶和内吞途径，并使用高分辨率的实时成像分析来可视化在两个模型系统中对受体运输的影响。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洞察力：1)胚胎发育中的命运决定；2)上皮片分裂中极性的重建；3)干细胞或分化系的信号依赖行为。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势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.