Localized mitochondrial metabolic activity in Xenopus mesendoderm cells undergoing collective cell migration

爪蟾中内胚层细胞集体细胞迁移的局部线粒体代谢活性

• 批准号: 10751722
• 负责人: GUSTAVO GAVREL PACHECO
• 金额: $ 3.66万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10751722
• 关键词: Address; Adenosine Monophosphate; Adhesions; Adhesives; Automobile Driving; Behavior; Biomechanics; C cadherin; Carbon; Cattle; Cell Nucleus; Cells; Childhood; Chimeric Proteins; Complex; Congenital Abnormality; Cues; Disease; Dissociation; Dominant-Negative Mutation; Ectoderm; Embryo; Embryonic Development; Endoderm; Environment; Extracellular Matrix; Fellowship; Fibronectins; Focal Adhesions; Genetic; Germ Layers; Glycolysis; Heart; Human; Human Development; Image; Immunologic Surveillance; Individual; Infant Mortality; Integrins; Investigation; Knowledge; Laboratories; Life; Ligands; Link; Lysine; Mechanics; Mediator; Mentors; Mesoderm; Metabolic; Metabolism; Microtubules; Mitochondria; Monoclonal Antibodies; Morphogenesis; Movement; Multicellular Process; Neoplasm Metastasis; Neural Tube Defects; Normal tissue morphology; Oligonucleotides; Pathologic Processes; Pathology; Phenotype; Physical Function; Physicians; Plasma; Play; Process; Protein Kinase; Proteins; Resolution; Respiration; Role; Scientist; Signal Transduction; Spatial Distribution; Stress; Testing; Tissues; Traction; Transcript; Warburg Effect; Work; Xenopus; aerobic glycolysis; cancer cell; career; cell behavior; cell motility; cohort; collaborative environment; critical period; experimental study; extracellular; gastrulation; genetic regulatory protein; inhibitor; mechanical force; mechanotransduction; migration; mitochondrial metabolism; pharmacologic; polarized cell; recruit; rho GTP-Binding Proteins; spatiotemporal; supportive environment

PROJECT SUMMARY/ABSTRACT [30 LINES MAX] Biomedical advances over the past twenty years have reduced the infant mortality rate in the US throughout the 21st century. However, congenital malformations that arise from dysregulated morphogenesis remain the leading cause of US infant mortality. An especially critical period of morphogenesis is gastrulation, a period of dynamic cellular rearrangements that establish the three tissue germ layers of ectoderm, mesoderm, and endoderm in the embryo. One notable cellular rearrangement in gastrulation is the collective migration of the mesendoderm along the blastocoel roof. Our laboratory identified α5ß1 integrin adhesion to fibronectin as a critical regulator of mesendoderm motility in gastrulation. During collective mesendoderm migration, the front row of migrating cells (leader cells) establishes substantial traction stress on the fibronectin extracellular matrix through α5ß1 integrin that allows for cells in rows behind the leader cells (follower cells) to be pulled forward. While the morphogenetic movements of gastrulation have been characterized, the metabolic processes that support these energetically expensive movements remain poorly understood. The experiments proposed herein will aim to unveil the mechanisms that integrate mechanical and metabolic signals in collective mesendoderm motility. Preliminary live imaging of mitochondrial activity and localization in collectively migrating mesendoderm explants supports the hypothesis that α5ß1 integrin mechanotransduction is a potent inducer of mitochondrial activity and recruitment to adhesions. In Specific Aim 1, we will test the role of α5ß1 integrin in organizing mitochondrial activity and localization, the spatial distribution of metabolites, and rates of glycolysis and mitochondrial respiration. We will induce differential activation of α5ß1 integrin in mesendoderm explants and dissociated primary cells via various fibronectin fusion proteins and varied mechanical environments. We will also modulate α5ß1 integrin activation in the mesendoderm of intact embryos via integrin-targeted monoclonal antibodies. In Specific Aim 2, we will explore the effects of mitochondrial and metabolic dynamic regulators on collective mesendoderm migration using live, high spatiotemporal resolution imaging of mitochondrial activity, mitochondrial localization, the spatial distribution of metabolites, and the function of metabolic regulatory proteins within the context of α5ß1 integrin-driven collective mesendoderm migration. In summary, the experiments proposed will reveal how α5ß1 integrin adhesion and signaling impact the metabolic processes that fuel energetically expensive gastrulation movements. By expanding our knowledge of the mechanisms that engage and fuel tissue rearrangements in gastrulation, we will form a better understanding of processes that contribute to congenital malformations in humans. This proposed fellowship will be performed in a collaborative and supportive environment to develop the trainee’s scientific independence and prepare him for a career as an independent physician-scientist in pediatric pathology. The applicant will be mentored by a supportive dissertation mentor (sponsor) and expert dissertation committee. 1

项目总结/摘要[最多30行] 过去二十年来，生物医学的进步降低了美国的婴儿死亡率。 21世纪。然而，由形态发生失调引起的先天性畸形仍然是一种常见的疾病。 美国婴儿死亡率的主要原因。形态发生的一个特别关键的时期是原肠胚形成， 动态细胞重排，建立外胚层、中胚层和 胚胎中的内胚层。原肠胚形成中一个值得注意的细胞重排是细胞的集体迁移。 中内胚层沿着囊胚腔顶。我们的实验室鉴定了α 5 β 1整合素与纤连蛋白的粘附， 原肠胚形成过程中内胚层运动的关键调节因子。在中内胚层集体迁移过程中， 一排迁移细胞（前导细胞）在纤连蛋白细胞外基质上建立了大量的牵引应力 通过α 5 β 1整联蛋白，其允许前导细胞（跟随细胞）后面的行中的细胞被向前拉动。 虽然原肠胚形成的形态发生运动已被描述，但 支持这些耗费大量精力的运动的原因仍然知之甚少。提出的实验 本文旨在揭示机制，整合机械和代谢信号在集体 中内胚层运动线粒体活性的初步实时成像和集体定位 迁移中内胚层外植体支持α 5 β 1整联蛋白机械转导是一种有效的 线粒体活性的诱导剂和粘连的募集。在具体目标1中，我们将测试α 5 β 1的作用 整合素在组织线粒体活性和定位，代谢产物的空间分布，以及 糖酵解和线粒体呼吸。我们将诱导中内胚层中α 5 β 1整合素的差异活化 外植体和解离的原代细胞通过各种纤连蛋白融合蛋白和各种机械 环境.我们还将通过以下途径调节完整胚胎中内胚层α 5 β 1整合素的活化 整合素靶向单克隆抗体。在具体目标2中，我们将探讨线粒体和 使用实时高时空分辨率的集体中内胚层迁移的代谢动态调节剂 线粒体活性成像、线粒体定位、代谢物的空间分布以及 代谢调节蛋白在α 5 β 1整联蛋白驱动的集合中内胚层中的功能 迁移总之，所提出的实验将揭示α 5 β 1整合素粘附和信号传导如何影响 为原肠形成运动提供能量的代谢过程。通过扩大我们 了解原肠胚形成中参与和促进组织重排的机制，我们将形成一个更好的 了解导致人类先天性畸形的过程。拟议的研究金 将在一个合作和支持的环境中进行，以发展受训者的科学 独立性和准备他的职业生涯作为一个独立的医生，科学家在儿科病理学。的 申请人将由支持论文导师（赞助商）和专家论文委员会指导。 1