Regulation of Cranial Suture Development by TGF-B and FG

TGF-B 和 FG 对颅缝发育的调节

• 批准号: 6884840
• 负责人: ARUN K GOSAIN
• 金额: $ 13.19万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-10 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6884840
• 关键词: bone morphogenetic proteins; clinical research; craniosynostosis; developmental genetics; developmental neurobiology; enzyme linked immunosorbent assay; fibroblast growth factor; gene expression; green fluorescent proteins; growth factor receptors; human subject; inhibitor /antagonist; laboratory mouse; mesenchyme; organ culture; patient oriented research; plasmids; polymerase chain reaction; transfection; transforming growth factors

DESCRIPTION (provided by applicant): Craniosynostosis is a significant health issue in humans, yet the pathogenesis and the mechanism of craniosynostosis are still unsolved. Clinical and experimental studies have implicated Transforming Growth Factor-a 1 (TGF-beta1) and Fibroblast Growth Factor-2 in cranial suture fusion. There is evidence that the effect of one or both molecules on suture fusion may be further altered by noggin, a bone morphogenetic protein antagonist. Gain of function mutations in FGF receptors have been reported in numerous human craniosynostosis syndromes, with the majority of these syndromes implicating FGF receptors 2 (FGFR2). However, the precise role of TGF-beta1 and FGF-2 in cranial suture development has not been explained. We hypothesize that 1) during the period of posterior frontal (PF) suture development in the mouse, altered biologic activity of TGF-beta1 and FGF-2 regulates fusion of the overlying suture calvaria, and 2) expression of these molecules in the fused PF suture in mice parallels that in fused sutures from human children with congenital craniosynostosis. Alteration in FGFR2 and noggin may also contribute to alter biologic activity of TGF-Beta1 and FGF-2 and further regulate suture fusion. Specific aims are 1) To quantity gene expression of TGF-beta1, FGF-2, FGFR2, and noggin in mouse cranial tissues harvested from both in vivo and in vitro models. Additionally, protein levels of TGF- beta1, FDF-2, and noggin will be measured in culture media in vitro using ELISA. 2) To quantitate gene expression of these same growth factors and receptors harvested from human patients with cogenital craniosynostosis. We will harvest tissue from both a fused suture and a patent suture from children undergoing craniotomy as part of the standard treatment for craniosynotosis. These data will be used to help determine the relationship between the mouse and human cranial suture systems. 3) To modulate cranial suture fusion in the mouse model, in vitro by modulating the bioavailability of the growth factor and receptors described in Aim 1. We will add recombinant forms of each molecule or antibodies to these molecules to either induce fusion of the normally patent sagittal suture or block the normal fusion of the PF suture. 4) To investigate the possibility of in vitro plasmid DNA transfection of mouse cranial tissue. If successful, gene manipulation of the target growth factors and receptors may be used to regulate cranial suture development in future studies. 5) To gain adequate knowledge of molecular biology to develop into an independent clinician-scientist. This proposal will provide mechanistic insights into the regulation of cranial suture development, and help to relate mouse and human models of craniosynostosis.

描述（由申请人提供）：颅突变是人类的重大健康问题，但是颅骨突变病的发病机理和机制仍未解决。临床和实验研究意味着转化生长因子-A 1（TGF-BETA1）和成纤维细胞生长因子-2在颅缝合融合中。有证据表明，一个或两个分子对缝合融合的影响可能会因骨形态发生蛋白拮抗剂Noggin而进一步改变。已经报道了许多人类颅突变综合症中FGF受体中功能突变的增益，其中大多数综合征涉及FGF受体2（FGFR2）。但是，尚未解释TGF-BETA1和FGF-2在颅骨缝合发育中的确切作用。我们假设1）在小鼠后额叶（PF）缝合发育期间，TGF-BETA1和FGF-2的生物学活性改变了calvaria上覆缝合的缝合缝线的融合，以及2）这些分子在小鼠中与人类cran fused parection fused sutios的融合pf Suture的表达。 FGFR2和Noggin的改变也可能有助于改变TGF-BETA1和FGF-2的生物学活性，并进一步调节缝合融合。具体目的是1）从体内和体外模型中收获的小鼠颅骨组织中TGF-BETA1，FGF-2，FGFR2和Noggin的数量基因表达。另外，使用ELISA在体外测量TGF-BETA1，FDF-2和Noggin的蛋白质水平。 2）定量从人类颅颅神经病患者收获的这些相同生长因子和受体的基因表达。我们将从融合的缝合线和接受颅骨切开术的儿童的专利缝合线中收集组织，这是颅鼻症的标准治疗方法的一部分。这些数据将用于帮助确定小鼠和人类颅骨缝合系统之间的关系。 3）通过调节AIM 1中描述的生长因子和受体的生物利用度调节小鼠模型中的颅骨缝合融合。我们将添加每个分子的重组形式，或对这些分子的重组形式，以诱导正常的正常sagittal Suture或阻止正常的正常融合的正常融合。 4）研究小鼠颅内组织的体外质粒DNA转染的可能性。如果成功，则可以使用对目标生长因子和受体的基因操纵来调节未来的研究中的颅骨缝合线发育。 5）获得足够的分子生物学知识，以发展成独立的临床医生。该提案将提供有关调节颅缝合发育的机械见解，并有助于将小鼠和人类模型的颅突症。