MOLECULAR GENETICS OF HUMAN BMP-4 IN FOP

FOP 中人类 BMP-4 的分子遗传学

• 批准号: 2712450
• 负责人: FREDERICK Samuel KAPLAN
• 金额: $ 29.47万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-06-01 至 2001-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2712450
• 关键词: DNA binding protein; DNA footprinting; blood chemistry; bone development disorder; clinical research; developmental genetics; gel mobility shift assay; gene expression; gene mutation; genetic markers; genetic promoter element; genetic regulatory element; human genetic material tag; human subject; immunocytochemistry; linkage mapping; messenger RNA; nuclear runoff assay; osteogenesis; pathologic ossification; polymerase chain reaction; progressive myositis ossificans; protein structure function; regulatory gene

The embryogenesis and regeneration of the skeleton are complex developmental events dependent on the successful induction of endochondral osteogenesis. Little is known, however, about the genetic control of bone induction. Bone morphogenetic proteins (BMPs) are members of a highly conserved family of molecules involved in the regulation of embryonic pattern formation and osteogenesis. Recombinant human BMP-4 can induce the entire developmental program of endochondral osteogenesis in an ectopic site in a manner identical to that seen in the autosomal dominant genetic disorder, fibrodysplasia ossificans progressive (FOP). FOP is a progressively disabling condition characterized by congenital malformations of the toes and disordered temporal and spatial induction of endochondral osteogenesis at ectopic sites. BMP-4 messenger RNA and proteins uniquely over-expressed inlymphocytes from patients who have FOP. These data indicate that over expression of BMP-4, a potent bone-inducing morphogen, is associated with disabling ectopic osteogenesis in man. Tow related hypotheses provide the focus for our longterm goals; First, the molecular structure and function of the human BMP-4 gene provides fundamental insight into the genetic regulation of endochondral bone induction and pattern formation in humans; second, BMP-4 is the prime candidate gene for FOP; and the molecular structure and/or regulatory control of that gene is abnormal in patients who have FOP. To address these hypothesis, we intend to: 1. Define the regulatory regions of the human BMP-4 gene by reporter gene expression assays. 2. Identify DNA-binding proteins for the BMP-4 gene in normal and FOP cells by gel mobility shift and footprinting assays using nuclear protein extracts from FOP and non-FOP cells. 3. Examine the rate of BMP-4 transcript initiation and mRNA stability in FOP cells relative to control cells by nuclear run-on and transcription inhibition experiments. 4. Examine the expression of BMP type I and type II receptors in FOP cells by RT-PCR and immunohistochemistry. 5. Identify genetic markers that are closely linked to the BMP-4 gene locus and utilize these markers to establish or exclude genetic linkage of the BMP-4 gene with FOP. Analysis of the regulatory control of the human BPM-4 gene will foster that longterm goal of elucidating basic mechanisms of normal and disordered bone induction, and of designing rational molecular diagnostic and treatment strategies for a wide range of developmental disorders of the skeleton in humans.

骨骼的胚胎发生和再生是复杂的 发展事件取决于成功归纳的 内软骨成骨。 但是，关于 骨诱导的遗传控制。 骨形态发生蛋白 （BMP）是高度保守的分子家族的成员 参与胚胎模式形成的调节和 成骨。 重组人BMP-4可以诱导整个 异位内骨内生成的发展程序 与常染色体显性中所见的方式相同 遗传疾病，纤维肿瘤发育不全的渐进性（FOP）。 fop 是一种以先天性为特征的逐渐致残的疾病 脚趾的畸形以及无序的时间和空间 在异位部位诱导内软骨成骨。 BMP-4 信使RNA和蛋白质独特地表达 来自患有FOP的患者的无细胞。 这些数据表明 BMP-4的表达是一种有效的骨形态学， 与人体中的异位成骨作用有关。拖 相关的假设为我们的长期目标提供了重点；第一的， 人BMP-4基因的分子结构和功能 提供对遗传调节的基本见解 人类的软骨骨诱导和模式形成； 其次，BMP-4是FOP的主要候选基因。和 该基因的分子结构和/或调节控制是 患有FOP的患者异常。为了解决这些假设， 我们打算：1。定义人类BMP-4的监管区域 通过报告基因表达测定的基因。 2。识别DNA结合 凝胶中正常和FOP细胞中BMP-4基因的蛋白质 使用核蛋白提取物移动性转移和足迹测定 来自FOP和非FOP细胞。 3。检查BMP-4的速率 相对于FOP细胞的转录本启动和mRNA稳定性相对于 通过核奔跑和转录抑制控制细胞 实验。 4。检查BMP I型和II型的表达 RT-PCR和免疫组织化学中FOP细胞中的受体。 5。 识别与BMP-4基因紧密相关的遗传标记 基因座并利用这些标记来建立或排除遗传 BMP-4基因与FOP的链接。监管分析 控制人BPM-4基因将促进这个长期目标 阐明正常和无序骨的基本机制 诱导以及设计合理的分子诊断和 各种发育障碍的治疗策略 人类的骨骼。