ISCHEMIC SKIN FLAP SURVIVAL USING AAV-FGF2 AND AAV-VEGF 165

使用 AAV-FGF2 和 AAV-VEGF 观察缺血性皮瓣的存活情况 165

• 批准号: 7959652
• 负责人: Paul Liu
• 金额: $ 23.92万
• 依托单位: ROGER WILLIAMS HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-01 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7959652
• 关键词: Accounting; Address; Adoptive Transfer; American; Anatomy; Biology; Blood Vessels; Body part; Build-it; Cells; Cessation of life; Chairperson; Computer Retrieval of Information on Scientific Projects Database; Devices; Dose; FGF2 gene; Funding; Gene Silencing; Gene Transfer; Genes; Grant; Hemoglobin; Hospitals; Institution; Ischemia; Length; Liposomes; Location; Measures; Mediating; Mentors; Methods; Mission; Modeling; Operative Surgical Procedures; Perfusion; Plasmids; Plastic Surgeon; Platelet-Derived Growth Factor; Population; Proteins; Publishing; Reconstructive Surgical Procedures; Research; Research Personnel; Resources; Skin; Small Interfering RNA; Societies; Source; Stem cells; Surgical Flaps; Techniques; Technology; Testing; Time; Tissue Engineering; Tissue Survival; Tissue Viability; Tissues; Training; Transgenes; United States National Institutes of Health; Universities; Vascular Endothelial Growth Factors; Vascular blood supply; Viral; Work; Wound Healing; adeno-associated viral vector; clinically relevant; design; gene therapy; improved; injury and repair; novel; novel strategies; statistics; wound

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. There has been no change in the scope of this project. This project will develop a novel application for a recent technique within gene therapy in the field of reconstructive surgery. We propose to use adeno-associated viral vectors designed to cause infected cells to elaborate potent blood supply-building proteins, namely VEGF, PDGF, and FGF2. This enhanced vascular network appears to rescue ischemic tissue from death, allowing "flaps" (tissue transferred from one anatomic location to another for the purpose of closing a wound or reconstructing parts of the body) to be constructed of longer length, greater size, or greater reliability. Statistics compiled by the American Society of Plastic Surgeons (www.plasticsurgery.org) tracked over 5.2 million reconstructive surgeries in the US last year alone. In addition, this project is germane to the overall mission of bettering wound healing, and may be applicable to any situation of tissue ischemia. It builds upon earlier, published work of the applicant (P Liu), who, though currently Chairman of Surgery at Roger Williams Hospital, Providence, RI, has never been the recipient of competitive Federal funding except a T32 training grant. It is not mentored, but will rely on the critical input from collaborators at Brown University and Roger Williams skilled in those techniques new to the applicant. The specific hypothesis tested is: Engineering tissue with AAV-delivered angiogenic genes can improve survival of ischemic flaps derived from that tissue via recruitment of endothelial progenitor cells. In addition to testing the effects of each of the transgenes, our approach will take advantage of the greater efficiency of viral-mediated gene transfer to assess the combination of VEGF + FGF2, which, when delivered via liposome in plasmid form, was more effective than single gene therapy delivered the same way. We propose the following specific aims: 1). Maximize tissue survival in a flap model by optimizing the timing and dosing of angiogenic gene transfers using AAV vectors, and assess the effects of combining VEGF and FGF2 gene therapy. 2). Develop a mechanism of action to account for enhanced tissue survival. We expect the approach to be both efficacious and clinically relevant. Addressing Aim 2 will help answer a controversial issue in vascular biology, namely, where does the new blood supply in injury repair come from? We will utilize siRNA methods of gene silencing to help get at that answer, as well as localization technology (IVIS) and adoptive transfer of endothelial progenitor cell-enriched populations into the ischemic tissue. Lastly, a new portable spectroscopic device, the ViOptix probe, measuring spectral shifts in the near infrared spectrum of oxygenated hemoglobin as a function of perfusion, will help determine real time tissue viability.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 该项目的范围没有变化。 该项目将在重建手术领域中为基因疗法中最近的技术开发新的应用。我们建议使用腺相关的病毒载体，以引起感染细胞来详细说明有效的血液供应蛋白，即VEGF，PDGF和FGF2。这种增强的血管网络似乎可以从死亡中挽救缺血组织，从而使“襟翼”（组织从一个解剖位置转移到另一个位置，目的是关闭伤口或重建身体的部分），以更长的长度，更大的可靠性构建。仅去年美国，美国整形外科医师学会（www.plasticsurgery.org）编制的统计数据就追踪了超过520万次重建手术。此外，该项目是依赖改善伤口愈合的整体使命，并且可能适用于组织缺血的任何情况。 它建立在申请人（P Liu）的早期出版的工作基础上，尽管目前在RI的Roger Williams Hospital担任外科董事长，但除了获得T32培训补助金以外，他从未获得过竞争性联邦资金。它没有受到指导，而是依靠布朗大学合作者和熟练申请人的这些技术的罗杰·威廉姆斯的关键意见。测试的特定假设是：具有AAV递送的血管生成基因的工程组织可以通过募集内皮祖细胞来改善从该组织得出的缺血瓣的存活。除了测试每个转基因的作用外，我们的方法还将利用病毒介导的基因转移的效率更高，以评估VEGF + FGF2的组合，当通过质粒形式中脂质体传递时，它比单一基因疗法更有效。我们提出以下具体目标： 1）。通过使用AAV载体优化血管生成基因转移的时间和剂量，在皮瓣模型中最大化组织存活，并评估将VEGFF和FGF2基因治疗相结合的效果。 2）。开发一种作用机制来解释增强的组织存活率。 我们希望这种方法既有效又具有临床意义。 解决目标2将有助于回答血管生物学中有争议的问题，即，新的损伤修复中新的血液供应来自何处？我们将利用基因沉默的siRNA方法来帮助获得该答案，以及本地化技术（IVI）和内皮祖细胞细胞增强种群的产物转移到缺血组织中。最后，一种新的便携式光谱设备Vioptix Probe，测量了氧化血红蛋白的近红外光谱的光谱变化，将有助于确定实时组织可行性。