权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Molecular mechanisms mediating the soft tissue attachment to teeth

介导软组织附着到牙齿的分子机制

基本信息

批准号：
10588063
负责人：
Jill Helms A Helms
金额：
$ 70.61万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-02-20 至 2028-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10588063
关键词：
Address Adenoviruses Anatomy Biochemical Bone Morphogenetic Proteins Bromodeoxyuridine Cell Cycle Cell Cycle Kinetics Cell Differentiation process Cells Clinical Connective Tissue Data Dental Cementum Differentiation Antigens Disease Embryo Epithelial Attachment Epithelial Cell Junction Epithelial Cell Proliferation Epithelial Cells Epithelium Excision Exposure to Flow Cytometry Fluorescent in Situ Hybridization Formulation Gene Expression Profile Gene Expression Profiling Genes Gingiva Gingivectomy Goals Harvest Health Hemidesmosomes Heterogeneity Histology Homeostasis Human Immunoglobulin G Immunohistochemistry Infiltration Inflammatory Injury Jaw Knowledge Label Libraries Ligature Maintenance Mediating Modeling Molecular Mus Natural regeneration Operative Surgical Procedures Oral Oral cavity Pathway interactions Periodontal Diseases Periodontal Ligament Periodontitis Placebos Population Prevention Prevention strategy Proliferating RNA Probes Recombinants Regenerative capacity Reporter Research Personnel Role Series Signal Transduction Site Skeleton Sorting Source Stains Tamoxifen Testing Therapeutic Tissues Tooth structure Tracer WNT Signaling Pathway Wnt proteins Wnt-3A protein Work alveolar bone beta catenin epithelial stem cell epithelium regeneration experimental study inhibitor injury and repair innovation insight lipid nanoparticle new therapeutic target notch protein novel therapeutic intervention preservation protein distribution repair model restoration self-renewal soft tissue stem cell biomarkers stem cell niche stem cell self renewal stem cells targeted treatment therapeutic protein tooth surface wound healing

项目摘要

Project abstract Maintaining the junctional epithelium (JE) is of primary importance if preservation of the cementum, periodontal ligament (PDL), and alveolar bone is to be achieved. New insights into JE barrier functions came with our discovery of a Wnt- responsive stem cell niche in the JE. In this proposal, our goal is to characterize the cellular players, niche signals, and regulatory mechanisms that control and maintain the JE stem cell niche in health, and after damage or disease. AIM 1 experiments will first test whether Wnt/β-catenin signaling is necessary for JE maintenance. Pathway inhibition will be achieved in Axin2LacZ/+ mice using adenovirus expressing the soluble Wnt inhibitor Dkk1; controls will receive adenovirus encoding the Fc portion of immunoglobulin G. At defined intervals, quantitative analyses will assess endogenous Wnt/β- catenin signaling via Xgal staining; JE hemidesmosomal gene and protein distribution via quantitative immunohistochemistry (qIHC); JE and GE cell cycle kinetics by EdU/BrdU labeling; and inflammatory cell infiltration in connective tissues underlying the JE and GE by FACS. Second, whether Wnt/β-catenin signaling is necessary for JE regeneration will be determined by subjecting Wnt lineage tracer e.g., Axin2CreERT2/+;R26RmTmG/+ mice to partial gingivectomy followed by Ad-Dkk1/Ad-Fc delivery. Lineage-tracing and quantitative analyses will establish a relationship between Wnt- responsive cell progeny, cell cycle kinetics, hemidesmosomal gene and protein distribution, and regeneration of JE barrier functions. AIM 2 experiments will evaluate the ability of a stabilized formulation of WNT protein to regenerate a functional JE. In one injury-repair model the JE will be surgically excised; in a second model, JE breakdown will be triggered via a ligature-induced periodontitis; both will be carried out in Axin2LacZ/+ and Axin2CreERT2/+;R26RmTmG/+ mice. Delivery of the WNT therapeutic will be followed at multiple timepoints by quantitative analyses to assess re-establishment of JE barrier functions. AIM 3 experiments will characterize Wnt-responsive JE stem cells and their progeny. Wnt-responsive stem cell pools from adjacent gingival epithelium (GE) will serve as control. Axin2CreERT2/+;R26RmTmG/+ mice will be exposed to tamoxifen, followed by harvest of JE and GE tissues at defined timepoints. GFP+ cells will be sorted by flow cytometry. Gene expression profiling of GFP+ cells will focus stem cell and differentiation markers. Fluorescent in situ hybridization will confirm gene expression patterns using RNA probe libraries corresponding to stem cell markers, components of Wnt/β-catenin, Notch, and Bone Morphogenetic Protein (BMP) pathways. Collectively, this proposal promises to provide important new insights into the requirement for Wnt/β-catenin signaling in maintaining the JE stem cell niche; regulating JE and GE cell proliferation and differentiation; and influencing hemidesmosomal-mediated attachment to the tooth surface. In addition, it should resolve the current debate over the molecular identities of JE v. GE stem cell pools and their differentiation potential. The proposed work also has the potential to identify an innovative therapeutic strategy for rebuilding a damaged JE and thus open new avenues for the restoration of the soft tissue attachment following periodontal diseases.

项目摘要