CELL AND GENETICS APPROACHES TO ENAMEL BIOMIMETICS

牙釉质仿生学的细胞和遗传学方法

基本信息

批准号：
8106413
负责人：
Malcolm L. Snead
金额：
$ 38.38万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
依托单位国家：
美国
项目类别：
财政年份：
1998
资助国家：
美国
起止时间：
1998-08-01 至 2015-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8106413
关键词：
Ablation Ameloblasts Amino Acids Animal Model Animals Architecture Biological Biomimetics C-terminal Cell Cycle Cells Communicable Diseases Coupled Data Dental Enamel Dental caries Disease Ectoderm Enamel Formation Engineering Gene Expression Genes Genetic Genome Grouping Habits Human Knock-in Mouse Knock-out Knockout Mice Knowledge Link Maps Measures Minerals Mus N Domain Normal tissue morphology Outcome Pattern Pharmaceutical Preparations Production Property Protein Engineering Proteins Proteomics Research Role Site Structure Structure-Activity Relationship Technology Tertiary Protein Structure Tissues Transgenic Mice Trauma Two-Hybrid System Techniques Work Yeasts amelogenin base biomineralization design enamel matrix proteins extracellular functional genomics genetic manipulation homologous recombination insight knockout animal knockout gene mouse Ambn protein mouse genome nanoscale novel protein function public health relevance retinal rods success tool

项目摘要

DESCRIPTION (provided by applicant): Deciphering structure/function relationships underpins the acquisition of biomedical knowledge while providing the basis to create novel materials and drugs. A powerful tool for deciphering structure/function outcomes has been germline genetic manipulation in mice. In this application, we apply germline manipulation to link structure to function for the enamel matrix protein ameloblastin. Enamel is a composite bioceramic tissue with unique material properties that are owed to its mode of biological fabrication. Ameloblast cells create an extracellular enamel protein matrix that serves to control both crystallite habit and the organization of crystallite bundles, allowing thousands of nanoscale crystallites to be organized and grouped together under the control of a single cell. We hypothesize that the grouping and organization of the nanocrystallites by a single ameloblast cell is the outcome achieved by the function(s) of critical domain(s) within the ameloblastin protein. The ameloblastin protein has been described as being organized into at least two domains, an N'- and a C'- terminal domain [Iwata et al. '07], based on the finding that following cleavage the N'- and C'- ameloblastin domains are no longer found in the same physical site in the enamel rod. The N'-terminus is enriched around the rod boundaries, much as a sheath covers a knife blade, while the C'-terminal domain behaves quite differently. We hypothesize that the N'-terminus of ameloblastin is likely responsible for the cell-to-matrix interactions that maintain the highly patterned rod-to-interrod boundaries observed in enamel. We hypothesize that the C'-terminus is responsible for the protein-to-mineral interactions producing the enamel bioceramic tissue. Outcomes will be measured by changes to stereotypic enamel architecture and by analysis of the material properties of the enamel in the knockin condition compared to wildtype animals. The outcomes from this experimental strategy will contribute to our understanding of functional genomics and proteomics while furthering our understanding of the formation of the only ectoderm-derived biomineralized tissue in the vertebrate body. Preliminary data from our research team suggest that this knockin approach will yield novel insights into the structure/function relationship for the ameloblastin protein, the second most abundant protein contributing to enamel organic matrix assembly and biomineralization. PUBLIC HEALTH RELEVANCE: The function(s) for the second most abundant protein of the forming mammalian enamel matrix is not known. Here, we map the function(s) of ameloblastin protein domains to the production of the enamel matrix required to control enamel biomineralization. In performing this project we will create an animal model useful to study caries, the most prevalent-, infectious-disease of humankind and provide one more design specification for creating an enamel biomimetic, useful for restoring enamel lost to trauma and disease.

描述（由申请人提供）：解密结构/功能关系基于获得生物医学知识的获取，同时提供了创建新型材料和药物的基础。破译结构/功能结果的强大工具是小鼠的种系基因操纵。在此应用中，我们将种系操作应用于将结构链接到牙釉质基质蛋白氨基蛋白的功能。搪瓷是一种复合生物陶瓷组织，其独特的材料特性归功于其生物制造方式。成熟细胞产生一个细胞外搪瓷蛋白基质，用于控制结晶石习惯和结晶石束的组织，使成千上万的纳米级结晶石可以在单个细胞的控制下组织并组合在一起。我们假设单个成成母细胞细胞是纳米晶体的分组和组织是由蛋白质细胞蛋白中临界结构域（s）的功能获得的结果。蛋白质细胞蛋白被描述为至少组织到两个结构域，一个n'-和一个c'-末端结构域[Iwata等。 '07]，基于以下发现：裂解后N'-和C'- ameloboblastin域不再在搪瓷杆的同一物理位置中找到。 N'-ensinus围绕杆边界丰富，就像鞘覆盖了刀刀片一样，而C'-端域的行为却大不相同。我们假设蛋白质细胞蛋白的N'-末端可能是导致细胞到矩阵的相互作用，这些相互作用维持了在搪瓷中观察到的高度图案化的杆到交互边界。我们假设C'-末端是导致产生搪瓷生物陶瓷组织的蛋白质到时段相互作用。与野生型动物相比，将通过对刻板印度牙釉质结构的变化以及对搪瓷条件下搪瓷的材料特性的分析来衡量结果。这种实验策略的结果将有助于我们对功能基因组学和蛋白质组学的理解，同时我们对脊椎动物体内唯一的外胚层生物矿化组织的形成的理解。我们研究团队的初步数据表明，这种敲蛋白方法将产生对蛋白质细胞蛋白的结构/功能关系的新见解，纤维细胞蛋白是第二大最丰富的蛋白质，促成搪瓷有机基质组装和生物矿化。公共卫生相关性：尚不清楚形成哺乳动物搪瓷基质的第二大蛋白质的功能。在这里，我们绘制了蛋白蛋白蛋白结构域的功能，以控制控制搪瓷生物矿化所需的搪瓷基质。在执行该项目时，我们将创建一个可用于研究龋齿的动物模型，这是人类最普遍的传染性疾病，并提供了另一种设计规范，用于创建牙釉质仿生剂，可用于恢复牙釉质失去创伤和疾病。