Evaluation molekularer Mechanismen während der osteogenen Differenzierung in Vorläuferzellen des dentalen Follikels (DFV)

牙滤泡祖细胞/干细胞（dfc）可用于细胞治疗，但其成骨分化机制尚不清楚。在该资助项目前期的工作中，可以发现BMP2或地塞米松可以诱导dfc的成骨分化。EDX分析表明，dfc产生的生物矿物质与分化的间充质骨髓干细胞相当。由于分化后的dfc不表达成牙本质标志物，因此它们对于骨水泥母细胞或牙槽成骨细胞分化机制的研究也非常有意义。该项目的先前结果表明，dfc分化的途径与bmp无关。在全基因组表达研究中，转录因子（TF） ZBTB16被确定为评估推定的与bmp无关的分化机制的起点。然而，对ZBTB16的初步研究表明，ZBTB16诱导BMP2，而TF DLX3诱导ZBTB16。在本项目开始之前，DLX3已被确定为dfc成骨分化的推定因子。该项目的研究表明，TF DLX3通过bmp2反馈控制支持dfc的成骨分化。基于这些结果，该项目现在将重点关注与BMP2/DLX3反馈回路相关的因素/途径。这里，有证据表明DLX3调节TF EGR1、NOTCH和WNT信号通路，这些信号通路也与分化的dfc相关。此外，细胞外基质（ECM）蛋白也影响dfc的分化。更新提案现在将检查dfc成骨分化过程中已确定的分子过程的功能关系。在第一部分中，分析了与DLX3支持的差异化相关的项目子过程。本文将研究TF EGR1或生物过程或分子功能（如WNT信号通路）与TF DLX3支持的dfc成骨分化的关系。在该项目的第二部分，将研究ECM蛋白结合整合素及其相关信号通路的影响。这种影响应在区分发展中国家的分子机制水平上加以评价；尤其是在BMP2/ dlx3通路上。所获得的关于分化的详细知识将为牙周组织的发展提供新的见解，也可能改善dfc治疗用途的选择，例如骨增强手术。

Dental follicle progenitor/stem cells (DFCs) could be used for cellular therapies in the future, but little is known about their osteogenic differentiation mechanisms. In the previous work of the funded project it could be shown that the osteogenic differentiation of DFCs can be induced with BMP2 or dexamethasone. EDX analyses have shown that DFCs produce biominerals, which are comparable to those of differentated mesenchymal bone marrow stem cells. Since differentiated DFCs did not express dentinogenic markers, they are also very interesting for research on the mechanisms of cementoblasts or alveolar osteoblasts differentiation. Previous results of this project suggested a BMP-independent pathway for the differentiation of DFCs. In genome-wide expression studies, the transcription factor (TF) ZBTB16 was identified as a starting point for the evaluation of putative BMP-independent mechanisms of differentiation. However, initial studies with ZBTB16 revealed that ZBTB16 induces BMP2 while the TF DLX3 induces ZBTB16. DLX3 was already identified as a putative factor for the osteogenic differentiation of DFCs before the start of this project. Investigations of this project showed that the TF DLX3 supports the osteogenic differentiation of DFCs by a BMP2-feedback control. Based on these results, the project will now focus on factors/pathways that are associated with the BMP2/DLX3 feedback loop. Here, there is evidence that DLX3 regulates the TF EGR1, the NOTCH and WNT signaling pathways, which are also associated with differentiating DFCs. Moreover, extracellular matrix (ECM) proteins also influence the differentiation of DFCs. The renewal proposal will now examine the functional relationship of identified molecular processes during osteogenic differentiation in DFCs. In the first part of the project sub-processes are analyzed, which are associated with the DLX3 supported differentiation. Here, the TF EGR1 or biological processes or molecular functions such as the WNT signaling pathway will be examined for their relation to the TF DLX3 supported osteogenic differentiation of DFCs. In the second part of the project the influence of ECM protein binding integrins and their associated signaling pathways will be investigated. This influence should be evaluated on the level of molecular mechanisms in differentiating DFCs; especially on that of the BMP2/DLX3-pathway. The obtained detailed knowledge about the differentiation will provide new insights into the development of the periodontium and may also improve options for the therapeutic use of DFCs, e.g. procedures of bone-augmentation.