Mechanistic roles of the basement membrane mechanics in cellular morphodynamics and tissue patterning of the pre-gastrulating embryo

基底膜力学在原肠胚形成前细胞形态动力学和组织模式中的作用

基本信息

批准号：
10617195
负责人：
Dong-Yuan Chen
金额：
$ 2.53万
依托单位：
CALIFORNIA INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-01 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10617195
关键词：
3-Dimensional Acute Address Affect Animals Anterior Architecture Basement membrane Biological Biological Process Biology California Cells Cellular biology Collaborations Complex Defect Development Developmental Biology Diffusion Embryo Embryonic Development Endoderm Endoderm Cell Environment Epiblast Epithelium Event Fellowship Fertilization Fluorescent in Situ Hybridization Fostering Foundations Future Gene Expression Genes Genetic Goals Human Image Image Analysis In Situ In Situ Hybridization Infertility Knowledge Learning Malignant Neoplasms Mammals Maps Mechanics Mesenchymal Modeling Molecular Morphogenesis Mus Neoplasm Metastasis Organ Pathologic Processes Pattern Pattern Formation Persons Play Pregnancy Prevention Principal Investigator Process Property Reaction Research Role Route Signal Transduction Specific qualifier value Structure Techniques Technology Testing Tissues Training Visceral Work blastomere structure cancer diagnosis cancer prevention career cell behavior cell fate specification conditional knockout differential expression early pregnancy loss embryo cell embryo culture embryo tissue experimental study extracellular gastrulation genetic manipulation image processing imaging approach implantation insight interdisciplinary collaboration mammalian embryology mathematical model migration mortality multi-scale modeling natural Blastocyst Implantation novel strategies preservation quantitative imaging skills stem cells superresolution microscopy tool transcriptome transcriptomics

项目摘要

Project Summary/Abstract Establishing proper tissue-level architecture and patterning during early embryogenesis is crucial for a successful pregnancy. A high rate of mortality seen in human embryos during the first 2-3 weeks post-fertilization is a major cause of early pregnancy loss, yet the essential cellular, molecular, and mechanical changes remain poorly defined. During early post-implantation mammalian embryogenesis, an extra-embryonic epithelial layer, the visceral endoderm, plays an essential role in the symmetry-breaking event that specifies the anterior- posterior patterning of the epiblast. Specifically, a subset of the visceral endoderm cells, the anterior visceral endoderm, migrates toward one end of the future anterior-posterior axis to pattern the epiblast. Recent work from our lab reveals that the embryonic basement membrane plays an essential role in symmetry breaking and morphogenesis that sets the stage for gastrulation. However, how the extracellular mechanics modulates the collective cellular dynamics and instructs cell identities for pattern formation remains not known. Here we propose to determine how the embryonic basement membrane coordinates the collective cell behaviors and facilitates anterior-posterior specification for pattern formation. First, we will comprehensively map the cell behaviors of the entire visceral endoderm with imaging approaches and define the role of the basement membrane in anterior visceral endoderm migration through genetic perturbations in stem cell-derived embryo- like structures as well as in the natural embryos. Next, we will map the basement membrane mechanics and apply a single-cell transcriptomics approach to generate in situ cell fate maps of the pre-gastrulating embryos. We will functionally test how the basement membrane mechanics regulate cell identities by correlating the gene expression profiles with basement membrane architecture and validate the findings with basement membrane- perturbed embryos. Finally, we will implement multi-scale mathematical models that integrate cell identities and cell dynamics with the basement membrane mechanics to uncover mechanisms of pattern formation. Overall, these experiments will unveil functional roles of the extraembryonic environment in defining the cell identities of the pre-gastrulation embryos. These findings will inform subsequent studies in my future goals on the conserved roles of the basement membrane and the extraembryonic tissues in topologically distinct embryos seen in human. Training during my fellowship period will expand my skillsets toward studying the more complex biology of mammals and broaden my knowledge on new aspects of developmental biology. The research environment at California Institute of Technology will further foster interdisciplinary collaborations that allow the development of novel approaches to investigate early mammalian embryogenesis.

项目摘要/摘要在早期胚胎发生过程中建立适当的组织水平结构和图案化对于A至关重要成功怀孕。在施肥后的头2-3周内，人类胚胎的死亡率很高是早期怀孕早期丧失的主要原因，但是必要的细胞，分子和机械变化仍然存在定义不佳。在植入后早期哺乳动物胚胎发生期间，胚胎外皮层，内脏内胚层在对称性的事件中起着至关重要的作用，该事件指定了前前 - 层状的后构图。具体而言，内脏内脏细胞的一个子集，前内脏内胚层向未来前后轴的一端迁移，以对层细胞进行模拟。最近的工作从我们的实验室中发现，胚胎基底膜在对称性破裂和形态发生奠定了胃阶段。但是，细胞外力学如何调节集体细胞动力学和指示模式形成的细胞身份尚不清楚。在这里，我们建议确定胚胎基底膜如何坐标集体细胞行为并促进了图案形成的前后验规范。首先，我们将全面映射通过成像方法的整个内脏内胚层的细胞行为，并定义了地下室的作用内脏内胚层迁移的膜通过干细胞衍生的胚胎中的遗传扰动像结构以及天然胚胎一样。接下来，我们将绘制地下室膜力学和应用单细胞转录组学方法来生成固定前胚胎的原位细胞命运图。我们将在功能上测试地下膜力学如何通过相关的基因来调节细胞身份具有地下膜结构的表达谱，并用地下膜验证发现干扰的胚胎。最后，我们将实施多尺度的数学模型，以整合细胞身份和用地下膜力学的细胞动力学揭示了模式形成的机制。总体而言，这些实验将揭示出胚外环境在定义细胞中的功能作用术前胚胎的身份。这些发现将在我未来的目标中为随后的研究提供信息地下室膜的保守作用和拓扑不同胚胎中的胚外组织在人类中看到。在我的团契期间的培训将使我的技能集扩大到研究更复杂的哺乳动物的生物学，扩大了我对发育生物学新方面的了解。研究加利福尼亚理工学院的环境将进一步促进跨学科合作开发新的方法来研究早期哺乳动物的胚胎发生。