CSF-1 in Dental Biology

CSF-1 在牙科生物学中的应用

• 批准号: 7173911
• 负责人: SHERRY L ABBOUD-WERNER
• 金额: $ 26.3万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-03-05 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7173911
• 关键词: Address; Adenovirus Vector; Age; Albers-Schonberg disease; Amelogenesis; Amelogenesis Imperfecta; Anti-Sense Probes; Binding; Biology; Code; Complementary DNA; Complex; Defect; Dental; Dental Enamel; Dentin; Dentin Dysplasia; Dentin Formation; Dentinogenesis Imperfecta; Deposition; Development; Embryo; Failure; Flow Cytometry; Functional disorder; Gene Delivery; Gene Expression; Gene Targeting; Genes; Genetic; Goals; Growth; Histologic; Homeostasis; Human; Immunohistochemistry; In Situ Hybridization; Incisor; Injection of therapeutic agent; Lead; Macrophage Colony-Stimulating Factor; Mandible; Mediating; Membrane; Messenger RNA; Modeling; Molecular; Molecular Profiling; Morphology; Mus; Newborn Infant; Osteocalcin; Osteoclasts; Pattern; Phenotype; Physiological; Polymerase Chain Reaction; Preparation; Process; Protein Isoforms; Proteins; Recombinants; Replacement Therapy; Resolution; Rodent Model; Role; Scanning; Spectrum Analysis; Staining method; Stains; Testing; Therapeutic; Thymidine; Time; Tissues; Tooth Diseases; Tooth Fractures; Tooth eruption; Tooth structure; Transgenic Organisms; adenoviral-mediated; day; deciduous tooth; design; gene replacement; homologous recombination; mineralization; novel; novel therapeutics; osteoclastogenesis; postnatal; promoter; protein expression; research study; time interval; vector

DESCRIPTION: Tooth matrix proteins are tightly regulated and critical for matrix deposition, mineralization and maturation. Altered expression of these proteins leads to enamel and/or dentin defects. We have made the novel observation that macrophage colony stimulating factor (CSF-1), a factor essential for osteoclast-mediated tooth eruption, is also required for normal tooth matrix formation. In op/op mice, a thymidine insertion in the coding sequence of the CSF-1 gene results in a deficiency of both the soluble (s)and membrane bound (m) forms of CSF-1 that decreases osteoclasts and leads to osteopetrosis and failure of tooth eruption. Analysis ofop/op teeth show a combined enamel and dentin defect characterized by thin, poorly organized enamel and dentin dysplasia that was associated with aberrant expression of tooth matrix proteins. Transgenic op/op mice harboring either the sCSF-1 or mCSF-1 cDNAs under the control of the osteocalcin promoter have also been generated and show distinct tooth phenotypes. Expression of sCSF-1 almost corrected the dental defects in op/op mice, while mCSF-1 partially corrected the enamel defect and resulted in an amelogenesis imperfecta phenotype. The long-term goal of this proposal is to determine the role of CSF-1 in primary tooth formation and the mechanisms that mediate the differential effects of CSF-1 isoforms on tooth matrix formation. Our first hypothesis is that CSF-1 is required for normal tooth matrix protein expression and that CSF-1 deficiency causes aberrant expression of these proteins that leads to specific dental defects during development. To address this issue, the expression profile of tooth matrix proteins, histologic features, mineralization and structural integrity by SEM ofop/op teeth will be compared to that of wt during embryonic and postnatal growth. Analysis of the endogenous developmental pattern of sCSF- 1 and mCSF- 1 expression in wt mice using in situ hybridization will be performed to elucidate the potential role of these isoforms in regulating tooth matrix proteins. Our second hypothesis is that sCSF-1 and mCSF-1 differentially regulate tooth matrix protein expression that, in turn, mediate the distinct biologic effects of these isoforms on tooth matrix formation. For these experiments, the patterns of matrix protein expression, mineralization and structural integrity during development will be analyzed in parallel in op/op mice expressing either sCSF-1 or mCSF-1 and correlated with rescue of the tooth phenotype in these mice. We will also test the hypothesis that adenoviral-mediated gene targeting of sCSF-1 to teeth will ameliorate the dental defects in op/op and, perhaps, rescue the enamel defect in mCSF-l-expressing op/op mice. These studies should increase our understanding of the molecular mechanisms involved in sCSF-1 and mCSF-l-mediated tooth matrix formation and may suggest novel therapeutic strategies designed to restore enamel and dentin tissues in a variety of dental disorders including amelogenesis imperfect a and dentinogenesis imperfecta.

描述：牙齿基质蛋白受到严格的调控，对基质沉积、矿化和成熟至关重要。这些蛋白的表达改变会导致牙釉质和/或牙本质缺陷。我们发现巨噬细胞集落刺激因子(CSF-1)是破骨细胞介导的牙齿萌出所必需的因子，也是正常牙齿基质形成所必需的。在OP/OP小鼠中，csf-1基因编码序列中的胸苷插入导致可溶性(S)和膜结合型(M)csf-1的缺失，从而减少破骨细胞，导致骨化生和牙萌出失败。分析显示牙釉质和牙本质联合缺陷，其特征是牙釉质和牙本质发育不良，这与牙齿基质蛋白的异常表达有关。在骨钙素启动子控制下携带sCSF-1或MCSF-1 cDNA的转基因OP/OP小鼠也已被产生，并显示出明显的牙齿表型。SCSF-1的表达几乎纠正了OP/OP小鼠的牙齿缺陷，而MCSF-1则部分纠正了牙釉质的缺陷，并导致了釉质形成不全的表型。这一建议的长期目标是确定CSF-1在乳牙形成中的作用，以及介导CSF-1亚型对牙齿基质形成的不同影响的机制。我们的第一个假设是，CSF-1是正常牙齿基质蛋白表达所必需的，而CSF-1缺乏会导致这些蛋白的异常表达，从而导致发育过程中特定的牙齿缺陷。为了解决这一问题，我们将通过扫描电子显微镜观察牙齿基质蛋白的表达谱、组织学特征、矿化情况和结构完整性，并将其与wt牙齿在胚胎和出生后生长过程中的情况进行比较。我们将用原位杂交的方法分析sCSF-1和MCSF-1在wt小鼠体内表达的发育模式，以阐明这些异构体在调控牙齿基质蛋白中的潜在作用。我们的第二个假设是，sCSF-1和MCSF-1对牙齿基质蛋白的表达有不同的调节作用，进而介导了这些异构体对牙齿基质形成的不同生物效应。在这些实验中，将平行分析表达sCSF-1或mcsf-1的OP/OP小鼠发育过程中基质蛋白的表达、矿化和结构完整性的模式，并与这些小鼠的牙齿表型挽救相关。我们还将验证腺病毒介导的sCSF-1基因靶向牙齿可以改善OP/OP小鼠的牙齿缺陷，并可能挽救表达mCSF-L的OP/OP小鼠的釉质缺陷。这些研究将增加我们对scsf-1和mcsf-L介导的牙齿基质形成的分子机制的了解，并可能提出新的治疗策略，用于修复各种牙病的釉质和牙本质组织，包括釉质形成不全和牙本质形成不全。