Enamel atlas: systems-level amelogenesis tools at multiple scales

牙釉质图谱：多尺度的系统级釉质生成工具

基本信息

批准号：
10670353
负责人：
Derk Joester
金额：
$ 64.12万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
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：
2021-08-01 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10670353
关键词：
Address Affect Alleles Ameloblasts Amelogenesis Amelogenesis Imperfecta Atlases Benchmarking Biological Process Biology Biophysics Cell Separation Cell physiology Cells Chromosome Mapping Color Communities Complement Cre driver Defect Dental Enamel Dental caries Dentition Development Developmental Process Dimensions Disease Electrons Elements Enamel Formation Flow Cytometry Fluorescence Gene Expression Gene Expression Profile Gene Expression Profiling Generations Genes Genetic Genetic Transcription Glean Health Heterogeneity Human Incisor Intervention Knock-in Length Life LoxP-flanked allele Maturation-Stage Ameloblast Measurement Mechanics Methods Minerals Modeling Molecular Profiling Mus Mutant Strains Mice Nanostructures Nature Pathway interactions Peptide Hydrolases Peptides Phase Physiological Population Process Production Property Proteins Proteome Proteomics Psychological Impact Quality of life Reagent Regulation Reporter Reporter Genes Research Research Personnel Societies Specific qualifier value Structure System Techniques Tissues Tooth structure Translations Wild Type Mouse X-Ray Medical Imaging biophysical analysis biophysical properties cell behavior conditional knockout cost differential expression empowerment enamel matrix proteins gene function genetic approach in vivo innovation interest live cell imaging mass spectrometric imaging mechanical properties mineralization multimodality mutant next generation novel novel strategies rare condition self assembly single-cell RNA sequencing spatiotemporal tool transcriptome sequencing transcriptomics

项目摘要

ABSTRACT Enamel defects, whether congenital, acquired, or environmental in origin, are associated with a significant cost to society and also have profound psychological impacts. Despite significant progress over the last decade, the developmental process that gives rise to enamel, known as amelogenesis, remains poorly understood. We have identified at least two factors that have delayed progress, and which we propose to address in this application. One is that existing mouse reagents, which provide the primary model for understanding genetic regulation of amelogenesis, have deficiencies that hinder dissecting the mechanisms in vivo. Another challenge is that new information regarding the nanostructure and phase composition of enamel have begun to emerge that prior models did not take into account. The ability to access powerful new genetic approaches, “omics” techniques and materials characterization methods therefore creates unprecedented opportunities to generate sophisticated new tools that will help push amelogenesis research to the next level. We propose to take full advantage of these recent technical advances and of the complementary expertise of our team to create an integrated, multi-modal set of tools and reference materials. Specifically, we will generate innovative mouse reagents, including amelogenesis-stage specific Cre drivers, reporters and conditional knockout and knock-in models of key structural and proteolytic players that constitute the enamel matrix, which will enable a workflow to profile transcription (using RNA sequencing) and translation (using proteomics) at specific developmental stages, and even on a single cell basis (using single-cell RNA sequencing). Tissue and cell-level molecular profiling will be complemented by an in-depth characterization of the structure, composition, and mechanical properties of forming and mature enamel at overlapping length scales. By mapping gene expression, specifying local proteomes, and quantitatively assessing impact of the perturbations at each of these levels on the materials properties of enamel, we will create a platform that will empower amelogenesis researchers, help delineate mechanisms of disease, and lay the groundwork to enable the development of new approaches of intervention.

抽象的牙釉质缺陷，无论先天性，获得或起源环境都与巨大的成本相关向社会产生深远的心理影响。尽管在过去十年中取得了重大进展，但引起搪瓷的发育过程被称为休闲生成，仍然知之甚少。我们有确定了至少两个因素延迟进度的因素，并且我们建议在此应用程序中解决这些因素。一个是现有的小鼠试剂，它为理解的遗传调节提供了主要模型没有变性的缺乏，可以阻碍体内的机制解剖。另一个挑战是新的有关搪瓷的纳米结构和相位组成的信息已开始出现模型没有考虑到。访问强大的新遗传方法的能力，“ OMICS”技术因此，材料表征方法创造了前所未有的机会来产生复杂的新工具将有助于将无体发生的研究提升到一个新的水平。我们建议充分利用这些最近的技术进步和我们的团队创建一组集成的多模式工具和参考材料。具体来说，我们将生成创新的小鼠试剂，包括特异性CRE驱动器，记者和条件构成搪瓷矩阵的关键结构和蛋白水解玩家的敲除和敲门模型，该模型将启用工作流程以进行轮廓转录（使用RNA测序）和翻译（使用蛋白质组学）特定的发育阶段，甚至是单个单元格（使用单细胞RNA测序）。组织和细胞水平的分子分析将通过对结构，组成，的深入表征完成以及在重叠长度尺度上形成和成熟搪瓷的机械性能。通过映射基因表达，指定局部蛋白质组织，并定量评估扰动的影响搪瓷材料属性上的级别，我们将创建一个平台，以增强浅弱点的能力研究人员，帮助描绘疾病的机制，并为新的发展做出基础干预方法。