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调控，对于基质沉积、矿化和成熟至关重要。这些蛋白质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 (m) 形式的 CSF-1 缺乏，从而减少破骨细胞并导致骨硬化和牙齿萌出失败。对op/op牙齿的分析显示，牙釉质和牙本质联合缺损，其特征是牙釉质薄、组织不良，牙本质发育不良，与牙基质蛋白的异常表达有关。还产生了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，并显示出不同的牙齿表型。 sCSF-1 的表达几乎纠正了 op/op 小鼠的牙齿缺陷，而 mCSF-1 部分纠正了牙釉质缺陷并导致釉质形成不全表型。该提案的长期目标是确定 CSF-1 在乳牙形成中的作用以及介导 CSF-1 亚型对牙基质形成的差异影响的机制。我们的第一个假设是 CSF-1 是正常牙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。为了解决这个问题，将通过扫描电镜将ofop/op牙齿的牙基质蛋白表达谱、组织学特征、矿化和结构完整性与胚胎和出生后生长期间的wt进行比较。将使用原位杂交对野生型小鼠中 sCSF-1 和 mCSF-1 表达的内源发育模式进行分析，以阐明这些亚型在调节牙基质蛋白中的潜在作用。我们的第二个假设是 sCSF-1 和 mCSF-1 差异调节牙基质蛋白的表达，进而介导这些亚型对牙基质形成的独特生物效应。对于这些实验，将在表达 sCSF-1 或 mCSF-1 的 op/op 小鼠中并行分析发育过程中基质蛋白表达、矿化和结构完整性的模式，并与这些小鼠中牙齿表型的拯救相关。我们还将测试这样的假设：腺病毒介导的 sCSF-1 基因靶向牙齿将改善 op/op 的牙齿缺陷，或许还可以挽救表达 mCSF-1 的 op/op 小鼠的牙釉质缺陷。这些研究应该增加我们对 sCSF-1 和 mCSF-1 介导的牙基质形成的分子机制的理解，并可能提出新的治疗策略，旨在恢复各种牙科疾病（包括釉质生成不全和牙本质生成不全）中的牙釉质和牙本质组织。