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 [最多 30 行] 过去二十年的生物医学进步降低了美国各地的婴儿死亡率然而，由于形态发生失调而引起的先天性畸形仍然是21世纪的问题。形态发生的一个特别关键的时期是原肠胚形成时期，是美国婴儿死亡的主要原因。动态细胞重排，建立外胚层、中胚层和内胚层三个组织胚层胚胎中内胚层的一种值得注意的细胞重排是原肠胚的集体迁移。我们的实验室鉴定出 α5ß1 整合素与纤连蛋白的粘附是沿囊胚腔顶部的中内胚层。原肠胚形成过程中中内胚层运动的关键调节者，在集体中内胚层迁移过程中，前部。一排迁移细胞（前导细胞）在纤连蛋白细胞外基质上产生巨大的牵引应力通过 α5ß1 整合素，允许领导细胞（跟随细胞）后面的行中的细胞被向前拉。虽然原肠胚形成的形态发生运动已被表征，但代谢过程支持这些耗能巨大的运动的建议仍然知之甚少。本文旨在揭示将机械和代谢信号整合到集体中的机制线粒体活动和集体定位的初步实时成像。迁移的中内胚层外植体支持以下假设：α5ß1 整合素机械转导是一种有效的线粒体活性诱导剂和募集粘连。在具体目标 1 中，我们将测试 α5ß1 的作用。整合素在组织线粒体活性和定位、代谢物的空间分布以及代谢率中的作用我们将诱导中内胚层中 α5ß1 整合素的差异激活。通过各种纤连蛋白融合蛋白和各种机械作用，外植体和解离的原代细胞我们还将通过调节完整胚胎中内胚层中的 α5ß1 整合素激活。在特定目标 2 中，我们将探讨线粒体和整合素靶向单克隆抗体的影响。使用实时高时空分辨率对集体中内胚层迁移的代谢动态调节剂线粒体活性、线粒体定位、代谢物的空间分布以及线粒体的成像 α5ß1整合素驱动的集体中内胚层中代谢调节蛋白的功能总之，所提出的实验将揭示 α5ß1 整合素粘附和信号传导如何影响。通过扩大我们的能量消耗的原肠胚形成运动的代谢过程。了解原肠胚形成过程中参与和促进组织重排的机制，我们将形成更好的了解导致人类先天畸形的过程。将在协作和支持的环境中进行，以培养学员的科学能力独立并为他成为儿科病理学独立医师科学家的职业生涯做好准备。申请人将得到支持性论文导师（赞助商）和专家论文委员会的指导。 1