Key Modulators of Cementogenesis

牙骨质形成的关键调节剂

• 批准号: 7808660
• 负责人: Martha J Somerman
• 金额: $ 37.83万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-25 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7808660
• 关键词: Address; Affect; Age; Animal Model; Animals; Area; Behavior Control; Bone Diseases; Calcinosis; Cell physiology; Cells; Cementoblast; Cementogenesis; Cementum Formation; Cervical; Chronic Kidney Failure; Clinical; Collaborations; Data; Dental Cementum; Dental Pulp; Dentin; Dentinogenesis; Dentistry; Development; Disease; Electrons; Extracellular Matrix Proteins; Factor Analysis; Familial hypophosphatemic bone disease; Figs - dietary; Functional disorder; Gene Expression; Gene Proteins; Genetic Transcription; Grant; Health; Histologic; Homeostasis; Human; Hypophosphatasia; Hypophosphatemia; Image; Immunohistochemistry; In Situ Hybridization; In Vitro; Institutes; Investigation; Knowledge; Lasers; Link; Location; Maintenance; Medical Research; Metabolic Diseases; Metabolism; Minerals; Modeling; Molecular Profiling; Mutation; Natural regeneration; Odontoblasts; Osteocytes; Partner in relationship; Plant Roots; Postdoctoral Fellow; Property; RNA Interference; Regulation; Reporting; Research; Rickets; Role; Thick; Tissues; Tooth root structure; Tooth structure; X ray spectroscopy; base; bone sialoprotein; cell behavior; dentin matrix protein 1; in vitro Model; inorganic phosphate; insight; knock-down; mineralization; nanoindentation; parent grant; protein expression; public health relevance; receptor; repaired; skeletal tissue; therapy design; transcription factor

DESCRIPTION (provided by applicant): Inorganic phosphate (Pi) homeostasis is essential for normal development, maintenance, and repair of teeth and skeletal tissues and yet the mechanisms controlling Pi homeostasis remain elusive. Under the parent grant, we hypothesized that cells associated with formation of cementum and dentin, during both development and regeneration, are highly sensitive to changes in Pi homeostasis. For this competitive revision proposal, based on new data suggesting that the ratio of Pi/PPi locally dictates protein expression of bone sialoprotein (BSP) vs. dentin matrix protein-1 (DMP1) in cementum, an additional the hypothesis has been set forth: The ratio of tissue Pi/PPi regulates cementum mineralized tissue apposition as well as composition of cementum extracellular matrix proteins, with an added aim to address this hypothesis: Specific Aim 1; sub-Aim A: To determine the mechanism of Pi/PPi regulation of matrix mineralization and gene/protein expression during tooth root cementogenesis. Strategies include expanding our animal models via a new collaboration with Dr. Jose Luis Millan (Burnham Institute for Medical Research). Dr. Milllan will be providing expertise and also animals with altered Pi/PPi ratio, i.e. Akp2 KO (low Pi/PPi) and Enpp1 KO (high Pi/PPi), and we will be preparing double KO (Akp2; Ank KO and Akp2; Enpp1 KO models). Tissues obtained from these animals will be examined histologically to include analysis of gene/protein expression profiles of single and double KO models by in situ hybridization, immunohistochemistry, and laser capture/microarray. Further, RNA silencing will be used to knock down expression of Ank, Enpp1, and Akp2 in OCCM.30 cementoblasts in vitro to identify, by microarray analyses, factors responding to altered Pi/PPi and potentially regulating BSP/DMP1 expression. The focus will be on identifying Pi/PPi related receptors, transporters, and transcription factors. In addition to providing important basic information on mineralized tissue development, these studies will inform practices relevant to human health. Results from these studies will provide new insights for designing therapies aimed at regenerating periodontal tissues, as well as providing extremely critical knowledge on Pi/PPi metabolism related to maintenance of Pi homeostasis. Furthermore, data generated will provide new information applicable to treatment of Pi associated disorders such as hypophosphatasia (HPP), X-linked hypophosphatemia (XLH), autosomal dominant and recessive hypophospatemic rickets (ADHR, ARHR), familial tumoral calcinosis (FTC), and chronic kidney disease (CKD). All of these severe disorders relate to dysfunction of Pi/PPi homeostasis and feature disorders of bones and teeth. The studies here expand our collaborations outside of dentistry and also will enable us to hire additional staff and a postdoctoral fellow. PUBLIC HEALTH RELEVANCE: Results from these investigations will provide key information as to the modulators of phosphate homeostasis. These new insights will assist in designing therapies aimed at regeneration of periodontal tissues, as well as providing extremely critical knowledge on Pi/PPi metabolism related to disorders such as hypophosphatasia (HPP), X-linked hypophosphatemia (XLH), autosomal dominant and recessive hypophospatemic rickets (ADHR, ARHR), familial tumoral calcinosis (FTC), and chronic kidney disease (CKD). All of these severe disorders relate to dysfunction of Pi/PPi homeostasis and feature disorders of bones and teeth.

描述（由申请人提供）：无机磷酸盐（PI）稳态对于正常发育，维护和修复牙齿和骨骼组织至关重要，但是控制PI稳态的机制仍然难以捉摸。在父母的授予下，我们假设在发育和再生过程中与形成水泥和牙本质相关的细胞对PI稳态的变化高度敏感。对于这项竞争性修订建议，基于新数据表明，PI/PPI的比率局部决定骨骼唾液蛋白（BSP）与牙本质基质蛋白-1（DMP1）的蛋白质表达，在胶结中，该假设已设置为：STERITY pPI/PPI的比例是质量/PPI的分类，并构成了组织的分类，并构成了组织的分类。蛋白质，以解决这一假设的额外目的：特定目的1； Sub-aim A：确定牙根胶结发生过程中基质矿化和基因/蛋白表达的PI/PPI调节的机制。策略包括通过与Jose Luis Millan博士（Burnham医学研究所）的新合作来扩展我们的动物模型。 Milllan博士将提供专业知识，还提供具有改变PI/PPI比的动物，即AKP2 KO（低PI/PPI）和ENPP1 KO（高PI/PPI），我们将准备双KO（AKP2; ANK KO和AKP2; ANK KO和AKP2 KO; ENPP1 KO;从这些动物中获得的组织将在组织学上检查，以包括原位杂交，免疫组织化学以及激光捕获/微阵列分析单个和双KO模型的基因/蛋白质表达谱。此外，RNA沉默将用于击倒ANK，ENPP1和AKP2的表达。30胶囊细胞在体外，通过微阵列分析，通过微阵列分析，响应改变PI/PPI的因素，并可能调节BSP/DMP1表达。重点将放在识别与PI/PPI相关的受体，转运蛋白和转录因子上。 除了提供有关矿化组织开发的重要基本信息外，这些研究还将为与人类健康相关的实践提供依据。这些研究的结果将为设计旨在再生牙周组织的疗法提供新的见解，并为与维持PI稳态有关的PI​​/PPI代谢提供极为重要的知识。此外，生成的数据将提供适用于治疗PI相关疾病的新信息，例如低磷酸（HPP），X连锁性低磷酸盐血症（XLH），常染色体显性症和隐性低体性rickeT（ADHR，ARHR，ARHR），ARDHR，ARHR，ARHR，ARHR，ARHR，家族性肿瘤（FTC）和家族性肿瘤病（FTC）和Chronic Sneyseys（CKD）和CKD（CKD）（CKD）。所有这些严重的疾病都涉及PI/PPI稳态功能障碍以及骨骼和牙齿的特征障碍。这里的研究将我们的合作扩展到牙科以外，还将使我们能够雇用更多的员工和博士后研究员。 公共卫生相关性：这些调查的结果将提供有关磷酸盐稳态调节剂的关键信息。这些新的见解将有助于设计旨在再生牙周组织的疗法，并提供与诸如下际磷酸盐（HPP），X连接的下磷酸血症（XLH），自身骨体显性占主导地位和衰落的低磷酸级别的抗磷酸菌（HPP）相关的PI/PPI代谢的极为重要的知识。 （FTC）和慢性肾脏疾病（CKD）。所有这些严重的疾病都涉及PI/PPI稳态功能障碍以及骨骼和牙齿的特征障碍。