Cell-specific regulation and downstream effects of Pitx2 and Eda expression in tooth initiation and replacement

Pitx2 和 Eda 表达在牙齿萌生和替换过程中的细胞特异性调控和下游效应

• 批准号: 10313900
• 负责人: Sophie L Archambeault
• 金额: $ 6.68万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2024-08-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10313900
• 关键词: Address; Adult; Affect; Alleles; Anhidrotic Ectodermal Dysplasia; Axenfeld-Rieger syndrome; Binding Sites; Bioinformatics; Biological Assay; Biology; Cell Proliferation; Cells; Cellular Morphology; Chromatin; Communities; Craniofacial Abnormalities; DNA Sequence; Dental; Development; Developmental Biology; Disease; Enhancers; Environment; Epithelial; Epithelial Cell Proliferation; Exhibits; Eye Abnormalities; Fishes; Gasterosteidae; Gene Activation; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Enhancer Element; Genetic Research; Genetic Transcription; Genomics; Goals; Hair; Heart Abnormalities; Histologic; Human; Hypodontia; In Situ Hybridization; Individual; Knowledge; Laboratories; Logic; Measures; Mediating; Mesenchyme; Methods; Modeling; Molecular; Morphogenesis; Mus; Mutation; Natural regeneration; Organogenesis; Outcome; Pathway interactions; Regulation; Regulator Genes; Research Institute; Signal Transduction; Signaling Molecule; Sweat Glands; Syndrome; System; Techniques; Testing; Time; Tissues; Tooth Abnormalities; Tooth Diseases; Tooth Germ; Tooth structure; Training; United States; Vertebrates; Work; Zebrafish; animal facility; cell behavior; cell type; craniofacial disorder; genetic manipulation; genome-wide; improved; loss of function; loss of function mutation; mouse model; mutant; novel; pleiotropism; response; skill acquisition; transcription factor; transcriptomics

PROJECT SUMMARY Dental and craniofacial abnormalities are some of the most common congenital conditions in the United States, and tooth abnormalities occur in 3-4% of adults. For several dental and craniofacial disorders, mutations in regulatory genes have been identified. For example, mutations in PITX2 and EDA are both associated with missing or misshapen primary and adult teeth. Understanding how these genes are regulated and function in normal and disease contexts is thus important to better understand how teeth form and regenerate. However, because developing teeth are dynamic, heterogeneous tissues, and the initiation of and response to molecular signals is often cell-type-specific, we currently lack a clear understanding of how Pitx2 and Eda are spatially and temporally regulated during tooth initiation, and which gene network(s) they affect to promote tooth placode proliferation. Therefore, the overall goal of this work is to determine the cell-type and tissue-specific gene regulation and signaling networks that underlie tooth placode initiation and morphogenesis. This goal will be achieved through the completion of several objectives in the threespine stickleback, a tractable fish model for developmental biology that, like humans but unlike mice, has the ability to replace teeth. First, genomic enhancers that regulate the expression of Pitx2 and Eda in developing teeth will be identified and analyzed to improve our ability to predict enhancers from genomic sequence (Specific Aim 1). Next, the effects of Pitx2 and Eda mutant alleles on early tooth germ cell proliferation and morphology will be quantified, both in initial and replacement teeth (Specific Aim 2). Finally, the downstream transcriptional effects of Pitx2 and Eda mutant alleles will be quantified using several complementary techniques, specifically in situ hybridization and cell proliferation assays (Specific Aim 2). This work will directly test the hypothesis that Pitx2 and Eda regulate dental epithelial cell proliferation of neighboring cells through distinct downstream targets. In addition to improving our functional understanding of two important master regulator genes that cause human tooth disorders, this work will provide training in traditional and cutting-edge developmental biology techniques in a dynamic and supportive scientific environment. The acquisition of these skills, interactions within the local scientific community, and training through courses at the research institute will facilitate the ability of the applicant to conduct rigorous, independent research of the genetic and developmental basis of tooth formation.

项目总结