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：
10467066
负责人：
Derk Joester
金额：
$ 63.65万
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/10467066
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 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 Maturation-Stage Ameloblast Measurement Mechanics Methods Microscopy 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 ameloblastin biophysical analysis biophysical properties cell behavior conditional knockout cost differential expression enamel matrix proteins gene function genetic approach in vivo innovation interest live cell imaging mass spectrometric imaging mechanical properties multimodality mutant next generation novel novel strategies rare condition self assembly single-cell RNA sequencing spatiotemporal tool transcriptome sequencing transcriptomics vibration

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，无论是先天性、后天性还是环境原因，都会带来巨大的成本尽管过去十年取得了重大进展，但我们对牙釉质形成的发育过程（称为釉质形成）仍知之甚少。确定了至少两个延迟进展的因素，我们建议在本申请中解决这些因素。一是现有的小鼠试剂，它为理解基因调控提供了主要模型釉质形成存在缺陷，阻碍了体内机制的解剖。另一个挑战是新的。关于牙釉质纳米结构和相组成的信息已经开始出现，之前模型没有考虑获得强大的新遗传方法、“组学”技术的能力。因此，材料表征方法创造了前所未有的机会来生成复杂的新工具将有助于将釉质形成研究推向新的水平。我们建议充分利用这些最新的技术进步和互补的专业知识我们的团队将创建一套集成的、多模式的工具和参考材料。创新的小鼠试剂，包括釉质形成阶段特定的 Cre 驱动程序、生产者和条件构成牙釉质基质的关键结构和蛋白水解参与者的敲除和敲入模型，将启用一个工作流程来分析转录（使用 RNA 测序）和翻译（使用蛋白质组学）特定的发育阶段，甚至在单细胞基础上（使用单细胞 RNA 测序）。细胞水平的分子分析将得到结构、组成、通过绘制基因图谱来确定重叠长度尺度上形成和成熟牙釉质的机械特性。表达，指定局部蛋白质组，并定量评估每个扰动的影响牙釉质材料特性的水平，我们将创建一个平台，将促进釉质生成研究人员帮助描绘疾病机制，并为开发新药物奠定基础干预方法。