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

基本信息

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
项目类别：
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

项目摘要

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

项目摘要/摘要[30行最大] 在过去的二十年中，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美国婴儿死亡率的主要原因。形态发生的特别关键时期是胃病，一个时期动态细胞重排建立了外胚层，中胚层和胚胎中的内胚层。胃的一个值得注意的细胞重排是沿Blastocoel屋顶的中胚层。我们的实验室确定α5ß1整合素对纤连蛋白的粘合剂为A 胃病中胚层运动的关键调节剂。在集体膜胚层迁移期间，正面一排迁移细胞（领导细胞）在纤连蛋白外基质上建立了重大的牵引力通过α5ß1整联蛋白，可以使领导细胞后面的行中的细胞向前拉。尽管已经表征了胃的形态发生运动，但代谢过程支持这些本质上昂贵的运动仍然知之甚少。提出了实验此处将旨在揭示集体中整合机械和代谢信号的机制中胚层运动。线粒体活性和本地化的初步实时成像迁移的中胚层外植体支持α5ß1整合素机械转导的假设是一种有效的线粒体活性和对依从性的诱因。在特定的目标1中，我们将测试α5ß1的作用整联蛋白在组织线粒体活性和定位，代谢物的空间分布以及速率糖酵解和线粒体呼吸。我们将诱导α5ß1整联蛋白在中胚层中的差异激活外植体和分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。通过靶向整联蛋白的单克隆抗体。在特定目标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胚层迁移的代谢动态调节剂线粒体活性，线粒体定位，代谢产物的空间分布以及代谢调节蛋白的功能在α5ß1整合蛋白驱动的集体中胚层的背景下迁移。总而言之，提出的实验将揭示α5ß1整合素的粘合剂和信号传导如何影响代谢过程有效地燃油昂贵的过口速度。通过扩展我们的了解参与和为胃内加油组织重排的机制，我们将形成更好的了解导致人类先天性畸形的过程。这个拟议的奖学金将在协作和支持的环境中进行，以发展受训者的科学独立并为他做好准备，成为小儿病理学独立身体科学家的职业。申请人将由支持的论文导师（赞助商）和专家论文委员会召集申请人。 1