Cell and mechanobiology of Asymmetric Cell Division

不对称细胞分裂的细胞和力学生物学

基本信息

批准号：
10793861
负责人：
Clemens C Clemens C. Cabernard
金额：
$ 23.83万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-01-01 至 2027-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10793861
关键词：
Acute Affect Behavior Cell Size Cell division Cells Central Nervous System Centrosome Chromatin Cues Cytoskeleton Defect Development Drosophila genus Generations Immunohistochemistry Interphase Kinesin Maintenance Malignant Neoplasms Medical Microtubule-Organizing Center Mitosis Molecular Myosin Type II Neurodevelopmental Disorder Organ Organelles Organism Pathway interactions Process Proteins RNA Reproducibility Research Siblings Sister Sister Chromatid Stereotyping System Tissues Transcriptional Regulation fly in vivo in vivo imaging innovation interest live cell imaging mechanical signal nanobodies nerve stem cell neuroblast non-muscle myosin novel optogenetics programs protein kinase N public health relevance rho GTP-Binding Proteins segregation spatiotemporal stem cells superresolution microscopy tool transcriptome sequencing tumor

项目摘要

PROJECT SUMMARY Generating cells with different fates, functions and behaviors is critically important for the development and maintenance of tissues, organs, and multicellular organisms. Cellular diversity can be generated through Asymmetric Cell Division (ACD). Stem cells utilize ACD to create differentiating sibling cells while maintaining the stem cell in the process. In addition to the asymmetric partitioning of proteins or RNAs, other mechanisms such as mechanical cues, sibling cell size asymmetry or organelle asymmetry could potentially also contribute to binary cell fate decisions. Here, I propose to use asymmetrically dividing Drosophila neuroblasts, the neural stem cells of the developing fly central nervous system, to investigate the cell and mechanobiology of ACD in vivo. Recently, we discovered that Non-muscle Myosin II-dependent cortical flows, induced through both polarity- and spindle-dependent cues, are implicated in the generation of sibling cell size asymmetry. I will investigate how cortical flows are induced and modulated with spatiotemporal precision to achieve reproducible sibling cell size asymmetry. Our recent discovery of Protein Kinase N (PKN), and the Rho GTPase pathway as inducers of cortical flows will provide molecular entry points. I will also investigate how cell size asymmetry contributes to cell fate decisions, using RNA sequencing, immunohistochemistry, and long-term live cell imaging in vivo. A second project encompassed in this research direction is aimed at investigating the molecular mechanisms and function of molecular centrosome asymmetry, which is manifested in biased microtubule organizing center (MTOC) activity in interphase. We identified new proteins and mechanisms, such as Kinesins, Pp4 and dynamic centriolar protein localization in mitosis, regulating centrosome asymmetry. Centrosome segregation is highly stereotypic in stem cells, but whether and how centrosome asymmetry affects cell fate decisions, remains to be resolved. We will use fly neural stem cells to investigate the mechanisms and functions of centrosome asymmetry during ACD. I am particularly interested in investigating whether centrosome asymmetry provides a mechanism for biased cell fate determinant segregation, either via asymmetric RNA or sister chromatid segregation. I will also investigate whether biased MTOC activity impacts transcriptional regulation via chromatin organization. This research program will benefit from several novel and innovative tools, consisting of live cell imaging, superresolution microscopy, RNA sequencing and acute protein mislocalization and perturbation systems (nanobody, optogenetics), which my lab implemented to probe cytoskeletal dynamics with high spatial and/or temporal precision in vivo. ACD is an evolutionary conserved mechanism and the proposed research program is medically significant because defects in ACD can cause neurodevelopmental disorders or cancer.

项目摘要产生具有不同命运，功能和行为的细胞对发展至关重要维持组织，器官和多细胞生物。蜂窝多样性可以通过不对称细胞分裂（ACD）。干细胞利用ACD创建不同的同胞细胞，而在此过程中保持干细胞。除了蛋白质或RNA的不对称分区之外，其他机理，例如机械提示，同胞细胞大小不对称或细胞器不对称性也有可能有助于二元细胞命运决策。在这里，我建议使用不对称分裂的果蝇神经细胞，即发展中枢神经系统，研究体内ACD的细胞和机械生物学。最近，我们发现非肌肉肌球蛋白II依赖性皮质流是通过极性和主轴依赖性提示与同胞细胞大小不对称的产生有关。我将调查如何皮层流是通过时空精度诱导和调节的，以实现可重复的同胞细胞大小不对称。我们最近发现的蛋白激酶N（PKN）和Rho GTPase途径作为诱导剂皮质流的含量将提供分子入口点。我还将研究细胞大小不对称的贡献在体内使用RNA测序，免疫组织化学和长期活细胞成像，以进行细胞命运决定。涵盖该研究方向的第二个项目旨在研究分子分子中心体不对称的机理和功能，该不对称体现在偏置的微管中相间的组织中心（MTOC）活性。我们确定了新的蛋白质和机制，例如驱动蛋白，PP4和动态中心蛋白在有丝分裂中定位，调节中心体不对称。中心体分离在干细胞中是高度定型的，但是否以及如何中心体不对称会影响细胞命运的决策，尚待解决。我们将使用蝇神经干细胞研究ACD期间中心体不对称的机制和功能。我特别感兴趣在研究中心体不对称是否提供了偏置细胞命运决定因素的机制通过非对称RNA或姐妹染色单体隔离分离。我还将调查是否有偏见 MTOC活动会通过染色质组织影响转录调节。该研究计划将受益于几种新颖和创新的工具，包括现场单元成像，超分辨率显微镜，RNA测序和急性蛋白质错误定位和扰动系统（纳米遗传学，光遗传学），我的实验室实施了该实施，以探测具有高空间和/或的细胞骨架动力学时间精度在体内。 ACD是一种进化保守的机制，拟议的研究计划是医学上的重要的是因为ACD缺陷会导致神经发育障碍或癌症。