Role of modulators of Pi/PPi in cementum formation and regeneration

Pi/PPi 调节剂在牙骨质形成和再生中的作用

基本信息

批准号：
10687850
负责人：
Emily Yin Chu
金额：
$ 24.9万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
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：
2021-09-14 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10687850
关键词：
Address Adult Affect Alkaline Phosphatase Animal Model Ankylosis Biological Assay CRISPR/Cas technology Cell Line Cementoblast Cementogenesis Cementum Formation Clinical Collaborations Coupled Data Defect Dental Cementum Dentin Development Development Plans Diagnostic Diphosphates Disease Exhibits Extracellular Matrix Extracellular Matrix Proteins Extracellular Space Fluorochrome Genes Glycoproteins Goals Growth Harvest Histology Hydroxyapatites Hypercementosis Immunohistochemistry In Situ Hybridization In Vitro Individual Institution Integrin Binding Intracellular Transport Knock-out Knowledge Label Laboratories Ligand Binding Link Maintenance Maps Mentorship Metabolism Methods Modeling Molecular Mus Mutant Strains Mice Mutation Natural regeneration Operative Surgical Procedures Pathogenesis Pathway interactions Periodontal Diseases Periodontal Ligament Periodontium Phenotype Prevention strategy Process Productivity Proteins Proteomics Regulation Research Research Personnel Rickets Role SCID Mice Scientist Serology Signal Pathway Site Solid Source Techniques Therapeutic Therapeutic Intervention Thinness Tissues Tooth Exfoliation Tooth Loss Tooth root structure Tooth structure Training Translating alveolar bone bone bone sialoprotein calcification career career development clinical application evidence base extracellular in vivo inhibitor inorganic phosphate insight member microCT mineralization mutant mutant mouse model osteopontin phosphoric diester hydrolase plasma cell membrane glycoprotein PC-1 premature prognostic assays protein expression psychosocial public health relevance pyrophosphatase regenerative therapy skeletal abnormality skills soft tissue targeted treatment therapy development training opportunity transcriptome sequencing

项目摘要

ABSTRACT Proper pyrophosphate regulation is critical in mineralization processes. Pyrophosphate (PPi) is a direct inhibitor of hydroxyapatite crystal growth, thus inhibiting mineralization. A main source of PPi is ATP, which is hydrolyzed by ectonucleotide pyrophosphatase phosphodiesterases (ENPP), including ENPP1. ENPP1 is expressed in bone along with progressive ankylosis protein (ANK), which transports intracellular PPi to extracellular spaces. While ANK and ENPP1 increase extracellular PPi, tissue non-specific alkaline phosphatase (TNAP, encoded by ALPL), hydrolyzes PPi into inorganic phosphate (Pi), thus decreasing PPi in extracellular spaces. Although the specifics of direct or indirect interactions between ANK, ENPP1, and TNAP are unknown, loss of any of these factors has dramatic effects on mineralized tissue. The studies proposed here were developed to elucidate functions of pyrophosphate regulators with the ultimate goal of developing therapies targeted toward mineralization disorders. Abnormal PPi regulation (as exhibited by individuals with ENPP1, ANK or ALPL mutations) can lead to alterations in the periodontium (cementum, periodontal ligament, alveolar bone), specifically cementum. Alterations include excessive cementum formation (ENPP1, ANK) or deficient cementum formation leading to premature tooth exfoliation (ALPL), an adverse sequelae of untreated periodontal disease. Periodontal disease, a global burden with significant psychosocial and financial consequences, features destruction of the periodontium. Existing regenerative therapies are unpredictable and do not target cementum regeneration. To investigate roles of Ank and Enpp1 during cementogenesis, I, along with my colleagues, developed Ank, Enpp1 double mutant mice. In preliminary studies, we noted a more dramatic hypercementosis phenotype in 8wk old Ank, Enpp1 double mutants versus single mutants. Additionally, we noted distinct differences between single and double mutants within the PDL region/cementum in gene/protein expression of SIBLING proteins, known modulators of the mineralization process. Three aims were developed to define ANK and ENPP1 roles in periodontal development and maintenance, identify mechanistic pathways in which ANK and ENPP1 regulate cementogenesis (e.g. focused on direct and indirect ability to modulate SIBLINGs), and determine the feasibility of using Enpp1 inhibitors to regenerate periodontaltissues. As a clinician scientist, my ultimate goal is to conduct productive research that provides scientific insights into periodontal disease pathogenesis and translate this knowledge into tangible clinical applications (e.g. diagnostic/prognostic tests, prevention strategies, and therapeutic interventions). My career development plan has been tailored toward this goal with solid mentorship, collaborations, and training opportunities in research and clinical settings. In conjunction with institutional support, I am confident studies/activities outlined in my application will help build upon my existing skillset and facilitate my transition into an independent investigator.

抽象的适当的焦磷酸盐调节在矿化过程中至关重要。焦磷酸盐 (PPi) 是一种直接羟基磷灰石晶体生长的抑制剂，从而抑制矿化。 PPi 的主要来源是 ATP，被外核苷酸焦磷酸酶磷酸二酯酶 (ENPP) 水解，包括 ENPP1。 ENPP1 是与进行性强直蛋白 (ANK) 一起在骨中表达，该蛋白将细胞内 PPi 转运至细胞外空间。当 ANK 和 ENPP1 增加细胞外 PPi 时，组织非特异性碱性磷酸酶（TNAP，由 ALPL 编码）将 PPi 水解为无机磷酸盐 (Pi)，从而降低 PPi 细胞外空间。尽管 ANK、ENPP1 和 TNAP 之间直接或间接相互作用的具体情况目前尚不清楚，任何这些因素的丧失都会对矿化组织产生巨大影响。提出的研究这里的开发是为了阐明焦磷酸调节剂的功能，最终目标是开发针对矿化障碍的疗法。 PPi 调节异常（如具有 ENPP1、ANK 或 ALPL 突变的个体所表现出的）可能导致牙周组织（牙骨质、牙周膜、牙槽骨），特别是牙骨质的改变。改变包括牙骨质形成过多（ENPP1、ANK）或牙骨质形成不足，导致牙齿过早脱落（ALPL），是未经治疗的牙周病的不良后遗症。牙周病疾病是一种全球性负担，具有严重的社会心理和经济后果，其特点是破坏牙周组织。现有的再生疗法是不可预测的，并且不针对牙骨质再生。到研究 Ank 和 Enpp1 在牙骨质形成过程中的作用，我和我的同事一起开发了 Ank， Enpp1双突变小鼠。在初步研究中，我们注意到更显着的牙骨质增生表型 8 周龄 Ank、Enpp1 双突变体与单突变体。此外，我们注意到之间的明显差异 SIBLING 蛋白基因/蛋白表达中 PDL 区域/牙骨质内的单突变体和双突变体，已知的矿化过程调节剂。制定了三个目标来定义 ANK 和 ENPP1 角色在牙周发育和维护中，确定 ANK 和 ENPP1 的机制途径调节牙骨质形成（例如，专注于调节 SIBLING 的直接和间接能力），并确定使用Enpp1抑制剂再生牙周组织的可行性。作为一名临床科学家，我的最终目标是进行富有成效的研究，为以下问题提供科学见解牙周病发病机制并将这些知识转化为切实的临床应用（例如诊断/预后测试、预防策略和治疗干预措施）。我的职业发展计划为实现这一目标而量身定制，提供扎实的指导、合作和研究培训机会和临床环境。结合机构支持，我对我的研究/活动充满信心申请将有助于增强我现有的技能，并促进我转变为独立调查员。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Orthodontic tooth movement alters cementocyte ultrastructure and cellular cementum proteome signature.

正畸牙齿移动改变牙骨质细胞超微结构和细胞牙骨质蛋白质组特征。

DOI：
发表时间：
2021-12
期刊：
影响因子：
4.1
作者：
Lira Dos Santos, Elis J;de Almeida, Amanda B;Chavez, Michael B;Salmon, Cristiane R;Mofatto, Luciana S;Camara;Tan, Michelle H;Kolli, Tamara N;Mohamed, Fatma F;Chu, Emily Y;Novaes, Pedro Duarte;Santos, Eduardo C A;Kantovit
通讯作者：
Kantovit

Cementocyte alterations associated with experimentally induced cellular cementum apposition in Hyp mice.

Hyp 小鼠中与实验诱导的细胞牙骨质沉积相关的牙骨质细胞改变。

DOI：
发表时间：
2021
期刊：
影响因子：
4.3
作者：
Lira Dos Santos, Elis J;Salmon, Cristiane R;Chavez, Michael B;de Almeida, Amanda B;Tan, Michelle H;Chu, Emily Y;Sallum, Enilson A;Casati, Marcio Z;Ruiz, Karina G S;Kantovitz, Kamila R;Foster, Brian L;Nociti Júnior, Francisco H
通讯作者：
Nociti Júnior, Francisco H