GENE THERAPY APPROACH FOR ENGINEERING CRANIOFACIAL BONE

工程颅面骨的基因治疗方法

• 批准号: 2907661
• 负责人: Renny Theodore Franceschi
• 金额: $ 22.71万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-03-01 至 2005-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2907661
• 关键词: Adenoviridae; bone morphogenetic proteins; bone regeneration; craniofacial; gene therapy; genetic promoter element; laboratory mouse; laboratory rabbit; nonhuman therapy evaluation; pathologic bone resorption; tissue engineering; transfection /expression vector

The long term goal of this project is to develop gene therapy as a viable approach for the tissue engineering of bone at oral and extraoral sites. Bone loss due to periodontal disease, neoplasia, trauma or reconstructive surgery is a major worldwide problem. Because current or emerging regenerative treatments using either bone grafts or recombinant proteins all have severe limitations, there is a need for new regenerative approaches. Gene therapy represents an exciting new alternative for stimulating bone formation. Genes for bone morphogenetic proteins (BMPs) are introduced into the patient at the desired site for bone regeneration. Recipient cells are then converted into mini-reactors to produce sustained levels of regenerative molecules. Preliminary studies with an engineered adenovirus expressing BMP7 support the feasibility of this approach in that this virus can induce ectopic bone formation at both subcutaneous and intramuscular sites. Two hypotheses will be tested in this project: 1) Host cells transduced with adenovirus vectors encoding bone morphogenetic proteins will produce sustained levels of regenerative molecules capable of inducing substantial new bone formation at both ectopic and orthotopic sites, 2) The BMP-driven regenerative process will recapitulate central events occurring in embyonic bone development and can be manipulated by altering the expression of other developmentally regulated factors. These hypotheses will be addressed by achieving the following specific aims: 1) Construct human adenovirus vectors expressing BMPs and optimize matrix carriers and conditions for in vivo viral delivery to cells, 2) Compare the osteogenic activity of adenovirus expressing BMPS 3, 4, and 7 alone and in combination using an in vivo mouse skin pouch model, 3) Examine the activity of optimized viral vector/matrix combinations showing strong osteogenic activity in Aim 2 for ability to form bone in both cranial and long bone critical size defect models, 4) Define early events following BMP virus transduction and manipulate the BMP response with known upstream and downstream modulators of BMP action. These studies will greatly advance our understanding of BMP-dependent bone regeneration and develop a viable gene therapy approach for regenerating bone in the craniofacial and appendicular skeleton.

该项目的长期目标是开发基因疗法，作为在口腔和口腔部位的骨骼组织工程的可行方法。 由于牙周疾病，肿瘤，创伤或重建手术而引起的骨质流失是全球的主要问题。 由于使用骨移植或重组蛋白的当前或新兴的再生治疗都有严重的局限性，因此需要采用新的再生方法。基因疗法代表了刺激骨形成的令人兴奋的新替代方法。 骨形态发生蛋白（BMP）的基因被引入所需部位的患者以进行骨骼再生。 然后将受体细胞转化为微型反应器，以产生持续水平的再生分子。 使用表达BMP7的工程腺病毒的初步研究支持这种方法的可行性，因为该病毒可以在皮下和肌内内部诱导异位骨形成。 该项目将测试两个假设：1）编码骨形态发生蛋白的腺病毒载体转导的宿主细胞将产生能够持续水平其他受发展调节因素的表达。这些假设将通过实现以下特定目的来解决：1）构建表达BMP的人类腺病毒载体并优化矩阵载体和体内病毒递送到细胞的条件，2）比较使用Vivo Mode a in In In vivo Mode a in In In vivo Mode a in In vivo Mode Spiririist，比较腺病毒的成骨活性组合在AIM 2中显示出强成骨活性，以便在颅骨和长骨临界大小缺陷模型中形成骨骼的能力，4）在BMP病毒转导后定义早期事件，并通过BMP动作的已知上游和下游调节剂来操纵BMP反应。这些研究将大大提高我们对BMP依赖性骨再生的理解，并开发一种可行的基因治疗方法，用于在颅面和阑尾骨骼中再生骨骼再生。