GENE TRANSFER INTO THE PULMONARY VASCULATURE

基因转移到肺血管中

• 批准号: 2029268
• 负责人: Elizabeth G Nabel
• 金额: $ 24.34万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-12-01 至 1998-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2029268
• 关键词: Adenoviridae; cell type; gene expression; immunocytochemistry; in situ hybridization; liposomes; lung; mature animal; newborn animals; polymerase chain reaction; pulmonary artery; pulmonary circulation; recombinant DNA; reporter genes; swine; tissue /cell culture; transfection; vascular endothelium

The study and treatment of many cardiopulmonary diseases are limited by the ability to express specific recombinant genes in nondividing cells in vivo. Gene transfer is the introduction of foreign DNA or gene sequences into host somatic cells, and this method facilitates in vivo investigations of gene expression and function. Delivery of recombinant genes into the pulmonary vasculature could potentially result in the expression of recombinant genes in large number of cells, including endothelial cells, type II pneumocytes, and distal airway epithelial cells. The expression of genes in these cells could facilitate investigations of the pathobiology of pulmonary vascular diseases and the potential development of gene therapy approaches to treat These vascular diseases. The purpose of this grant is to test the hypothesis that recombinant genes can be delivered and expressed in the pulmonary vasculature in vivo. In Specific Aim I, recombinant reporter genes will be introduced into the pulmonary vasculature using adenoviral and DNA liposome vectors, gene expression will be confirmed by analysis of DNA, mRNA and protein; cell types which are transduced will be defined; and the duration of gene expression in the lung and other tissues will be investigated. In Specific Aim II, modifications will be made in these vectors to optimize transfection efficiencies and gene expression in vivo. Lung specific gene expression will be developed using surfactant promoters. In Specific Aim III, the hypothesis that recombinant genes may persist longer in neonatal tissue compared with adult tissue will be tested by investigations into the stability of gene expression following gene transfer with adenoviral and DNA liposome vectors into neonatal and adult lungs. Preliminary studies in our laboratory demonstrate the feasibility of these studies. In a porcine animal model, we have shown that recombinant reporter genes can be expressed at specific sites in the pulmonary vasculature following transcatheter injection. In a human gene therapy phase I study, transcatheter delivery of HLA-B7 DNA and liposomes into a right posterior basal segment artery was safely performed in a patient with melanoma metastatic to the right lower lobe. The goal of this proposal is to optimize the delivery and expression of recombinant genes in the pulmonary vasculature following direct gene transfer with adenoviral and DNA liposome vectors. In addition, developmental differences in gene expression will be investigated in neonatal and adult lungs. The ability to express these recombinant genes at an early age may allow for early interventions designed to prevent the progression of pulmonary diseases. Taken together, these studies should provide insight into basic mechanisms of gene expression within pulmonary tissue and may provide an approach to the genetic treatment of pulmonary vascular diseases.

许多心肺疾病的研究和治疗受到 在非分明细胞中表达特定重组基因的能力 体内。基因转移是外源DNA或基因序列的引入 进入宿主体细胞，此方法促进体内 研究基因表达和功能。递送重组 进入肺脉管系统的基因可能会导致 重组基因在大量细胞中的表达，包括 内皮细胞，II型肺细胞和远端气道上皮 细胞。这些细胞中基因的表达可以促进 研究肺血管疾病的病理学和 基因治疗方法的潜在发展来治疗这些血管 疾病。这笔赠款的目的是检验以下假设 重组基因可以在肺部传递和表达 体内脉管系统。在特定目标I中，重组记者基因将是 使用腺病毒和DNA引入肺脉管系统 脂质体载体，基因表达将通过分析DNA证实 mRNA和蛋白质；转导的细胞类型将被定义；和 肺和其他组织中基因表达的持续时间将是 调查。在特定的AIM II中，将进行修改 在体内优化转染效率和基因表达的载体。 肺特异性基因表达将使用表面活性剂开发 发起人。在特定的目标III中，重组基因可能 与成人组织相比，在新生儿组织中持续更长的时间 通过研究基因表达的稳定性测试 用腺病毒和DNA脂质体向量转移基因转移到新生儿和 成人肺。我们实验室的初步研究表明 这些研究的可行性。在猪动物模型中，我们显示了 重组报告基因可以在特定位置表达 经导管注射后肺脉管系统。在人类基因 治疗I期研究，HLA-B7 DNA和脂质体的经导管递送 在A中安全地进行右后段动脉 患有黑色素瘤转移到右下叶的患者。目标的目标 建议是优化重组基因的递送和表达 在直接基因转移之后的肺脉管系统中 腺病毒和DNA脂质体载体。此外，发展 基因表达的差异将在新生儿和成人中研究 肺。在很小的时候表达这些重组基因的能力可能 允许旨在防止进展的早期干预措施 肺部疾病。综上所述，这些研究应提供洞察力 进入肺组织中基因表达的基本机制，可能 提供了一种肺血管遗传治疗的方法 疾病。