Primary Genes Promoting Cementoblast Differentiaton

促进成牙骨质细胞分化的初级基因

• 批准号: 6936009
• 负责人: SOTIRIOS TETRADIS
• 金额: $ 36.22万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-06-01 至 2008-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6936009
• 关键词: cell differentiation; cell line; cementum; clinical research; dental development; developmental genetics; human tissue; interleukin 1; laboratory mouse; laboratory rat; mitogen activated protein kinase; prostaglandin E; regeneration; subtraction hybridization; transcription factor

DESCRIPTION (provided by applicant): Periodontal regeneration is an important, yet unattainable, treatment objective in dentistry. Developing better regenerative treatments requires a deeper understanding of cementoblast, osteoblast, and PDL fibroblast biology. The recently established OC-CM cementoblastic cell line provides an invaluable research tool for studying cementoblast molecular biology. We found that prostaglandin (PG) E2 and fluprostenol (flup; a specific FP receptor agonist) promoted, while interleukin-1 (IL-1) inhibited, OC-CM mineralization and differentiation. These effects were reproducible in primary human cementoblasts. Mechanistically, receptor bound flup and IL-1 induce primary genes, which regulate downstream genes and ultimately control cell function. Some primary genes are induced by both flup and IL-I, while others are preferentially induced by each treatment. We hypothesize that preferentially induced primary genes dictate flup- and IL-l-regulated cementoblast function. To identify primary genes we performed subtraction hybridization using OC-CM cells treated with 0.1 (M flup or 10 ng/ml IL-1 for 90 min. Two cDNA subtractions were performed: (a) flup - IL-l, to identify flup-induced genes and (b) IL-1 - flup, to identify IL-l-induced genes. Several preferentially induced primary genes were identified. Our objective is to study the role of the transcription factors Nur77 and egr1, and the MAP kinase signaling regulator mkp1 in cementoblast function. In preliminary studies, mRNA from all three genes was rapidly and transiently induced by flup, but not by IL-1, in OC-CM cells. This temporal pattern of Nur77, egr1, and mkp1 mRNA expression was replicated in primary human cementoblasts and primary mouse osteoblasts treated with flup. We propose three Specific Aims: 1) to characterize Nur77, egr1, and mkp1 gene regulation by prostanoids in cementoblasts, 2) to characterize Nur77, egr1, and mkp1 protein involvement in regulating cementoblast gene expression and differentiation, and 3) to examine PGE2 and PGF2alpha induction of Nur77, egr1, and mkp1 gene expression in cementoblasts in vivo. We anticipate that Nur77, egr1, and mkp1 will be key mediators of cementoblast differentiation. These studies will contribute to our understanding of the molecular signals that regulate cementoblast function.

描述（由申请人提供）：牙周再生是牙科中一个重要但无法实现的治疗目标。开发更好的再生疗法需要更深入地了解成牙骨质细胞、成骨细胞和 PDL 成纤维细胞生物学。最近建立的 OC-CM 成牙骨质细胞系为研究成牙骨质细胞分子生物学提供了宝贵的研究工具。我们发现前列腺素 (PG) E2 和氟前列醇（flup；一种特定的 FP 受体激动剂）促进 OC-CM 矿化和分化，而白介素-1 (IL-1) 则抑制。这些效应在原代人成牙骨质细胞中可重现。从机制上讲，受体结合的 flup 和 IL-1 诱导初级基因，从而调节下游基因并最终控制细胞功能。一些主要基因由flup和IL-1两者诱导，而其他基因则由每种处理优先诱导。我们假设优先诱导的初级基因决定 flup 和 IL-1 调节的成牙骨质细胞功能。为了鉴定主要基因，我们使用用 0.1 (M flup 或 10 ng/ml IL-1 处理 90 分钟的 OC-CM 细胞进行消减杂交。进行了两次 cDNA 消减：(a) flup - IL-1，以鉴定 flup-诱导基因和（b）IL-1 - flup，以鉴定IL-1诱导基因。我们的目的是研究转录因子Nur77和的作用。在初步研究中，在 Nur77、egr1 的这种时间模式中，来自所有三个基因的 mRNA 均被 flup 快速而短暂地诱导，但不被 IL-1 诱导。 ，并且 mkp1 mRNA 表达在用 flup 处理的原代人成牙骨质细胞和原代小鼠成骨细胞中得到复制。我们提出了三个具体目标：1) 表征 Nur77，成牙骨质细胞中前列腺素类对egr1和mkp1基因的调节，2）表征Nur77，egr1和mkp1蛋白参与调节成牙骨质细胞基因表达和分化，3）检查PGE2和PGF2α对牙骨质细胞中Nur77，egr1和mkp1基因表达的诱导体内的成牙骨质细胞。我们预计 Nur77、egr1 和 mkp1 将成为成牙骨质细胞分化的关键介质。这些研究将有助于我们了解调节成牙骨质细胞功能的分子信号。