Intrinsic and extrinsic spindle orientation mechanisms in mammalian epidermis

哺乳动物表皮的内在和外在纺锤体定向机制

基本信息

批准号：
10360689
负责人：
SCOTT E WILLIAMS
金额：
$ 30.55万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
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：
2021-03-01 至 2026-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10360689
关键词：
Ablation Adherens Junction Adhesions Adhesives Anaphase Apical Architecture Basal Cell Behavior Bulla Cadherins Carcinoma Cell Density Cell Fate Control Cell division Cell-Cell Adhesion Cell-Matrix Junction Cells Communities Complex Cues Daughter Defect Development Disease Ectodermal Dysplasia Embryo Epidermis Epidermolysis Bullosa Epithelial Equilibrium Focal Adhesions GPSM1 gene GPSM2 gene Gene Expression Gene Silencing Genes Genetic Genetic Models Genetic Techniques Goals Growth Homeostasis Homologous Gene Human Hyperplasia Image Integrins Intrinsic factor Junctional Epidermolysis Bullosa Knowledge Label Lead Malignant Neoplasms Mediating Metaphase Mitosis Modeling Molecular Morphogenesis Mutate Mutation Nature Online Mendelian Inheritance In Man Outcome Pathogenesis Pathologic Pathway interactions Pattern Phenotype Phosphorylation Play Positioning Attribute Process Property Proteins Proteomics RNA Interference Resources Role Scaffolding Protein Shapes Signal Pathway Skin Skin Abnormalities Stratification Stratified Epithelium Stratum Basale Structure Syndrome Techniques Time Tissues Variant Vinculin Work afadin alpha catenin cell cortex cleft lip and palate cohort ex vivo imaging exhaustion experimental study flexibility human disease in vivo innovation insight knock-down loss of function mechanotransduction mouse model nectin paralogous gene progenitor protein protein interaction real-time images response self-renewal skin disorder skin organogenesis stem stem cell division stem cells telophase therapy development tissue stress tool treatment strategy ultrasound

项目摘要

ABSTRACT Proper control of stem cell division is critical for tissue morphogenesis and homeostasis. When dysregulated, it can lead to hypoplasia and stem cell exhaustion on the one hand, or tissue overgrowth and cancer on the other. But mitosis is more than simple proliferation, as cell division can be controlled not only in time but also in space. Oriented cell divisions (OCDs) are an example of the latter, and for stem and progenitor cells, choices between division axes can dictate cell fate outcomes and impact tissue architecture. In stratified epithelia such as the epidermis, basal progenitors divide either within the plane of the epithelium, or perpendicular to it. Evidence suggests that planar divisions are generally self-renewing symmetric cell divisions (SCDs) while perpendicular divisions are differentiative asymmetric cell divisions (ACDs). Previous work from our lab has shown that ACDs are directed by a complex of polarity and spindle orientation proteins—converging on the critical scaffolding protein LGN (Gpsm2)—which localize asymmetrically at the apical cell cortex. More recently, we have found that the paralog AGS3 (Gpsm1) seems to oppose LGN, and functions in promoting SCDs through an unknown mechanism. In addition, we recently made the surprising discovery that division orientation is not fixed during metaphase, as previously thought, but can be further refined during late stages of mitosis. In this process, which we term “telophase correction,” roughly one-third of basal cells enter anaphase at oblique angles, but then reorient to either planar or perpendicular. We have learned that cell-cell adhesions—specifically, the mechanosensing components of the adherens junction—are important for telophase correction to occur, and can operate independently of LGN. This demonstrates that in addition to intrinsic cues such as the LGN complex, extrinsic factors such as the local tissue microenvironment influence the final division axis. Despite what we and others have learned about the molecular control of ACDs, major knowledge gaps exist in understanding how oriented divisions shape tissue architecture both during normal development and in congenital skin diseases such as epidermolysis bullosa and ectodermal dysplasia. Specifically, the objectives of this proposal are to develop a better understanding of 1) what regulates SCDs and how the choice between SCD/ACD is made (SA1), 2) how cell-cell adhesion, cell-matrix, and local cell density impact division orientation and fate decisions (SA2). To achieve these goals, we will leverage a combination of innovative approaches, centered on our rapid, high-throughput technique—lentiviral ultrasound-guided gene inactivation and gene expression (LUGGIGE)— which we will utilize to generate mouse models of both gene loss and of specific mutations found in human diseases. Combined with ex vivo imaging of skin explants and in vivo proteomic approaches to characterize the LGN and AGS3 interactomes using TurboID, these comprehensive studies will provide insights into the cell- intrinsic and extrinsic cues that determine division orientation, and how they operate during normal epidermal growth and in blistering and dysplastic skin diseases.

抽象的正确控制干细胞分裂对于组织形态发生和稳态至关重要。当失调时，它一方面会导致发育不全和干细胞衰竭，另一方面会导致组织过度生长和癌症。但是有丝分裂不仅仅是简单的增殖，因为不仅可以及时而且在太空中控制细胞分裂。定向细胞分裂（OCD）是后者的一个例子，对于茎和祖细胞，选择分裂轴可以决定细胞脂肪的结果并影响组织结构。在分层的上皮中，例如证据，基本祖细胞在上皮平面内分裂，或垂直于其。表明平面分裂通常是自我更新的对称细胞划分（SCD），而垂直于分裂是区分不对称的细胞分裂（ACD）。我们实验室的先前工作表明ACD 由极性和主轴取向蛋白的复合物引导 - 临界脚手架蛋白LGN（GPSM2） - 在根尖细胞皮质中不对称地定位。最近，我们发现 Paratorog AGS3（GPSM1）似乎反对LGN，并通过未知来促进SCD 机制。此外，我们最近提出了令人惊讶的发现，即在如前所述，中期可以在有丝分裂的后期进一步完善。在此过程中，我们称“末期校正”，大约有三分之一的基本细胞以倾斜角度进入后期，但是重新定位平面或垂直。我们了解到细胞细胞的综述，特别是粘附连接的机械传感成分 - 发生末期校正很重要，并且可以独立于LGN操作。这表明，除了固有的线索（例如LGN复合物）外诸如局部组织微环境等外部因素会影响最终分裂轴。尽管我们和其他人了解了ACD的分子控制，在理解如何定向分裂在正常发育和先天性皮肤疾病中塑造组织结构例如表皮分解球囊和外胚层发育不良。具体而言，该提议的目标是更好地了解1）哪种调节SCD以及如何做出SCD/ACD之间的选择（SA1），2）细胞 - 细胞粘合剂，细胞 - 矩阵和局部细胞密度影响分裂方向和脂肪决策如何（SA2）。为了实现这些目标，我们将利用创新方法的结合，集中在我们的快速，高通量技术 - 居住超声引导的基因失活和基因表达（Luggige） - 我们将利用它来生成基因损失和人类特异性突变的小鼠模型疾病。结合皮肤外植体和体内蛋白质组学方法的体内成像，以表征 LGN和AGS3使用涡轮增压体相互作用，这些全面的研究将为细胞提供见解。确定分裂方向的固有和外在提示，以及它们在正常表皮期间的运作方式生长，发脾气和发育不良的皮肤疾病。