GENE TRANSFER INTO THE PULMONARY VASCULATURE

许多心肺疾病的研究和治疗受到限制， 在非分裂细胞中表达特定重组基因的能力， vivo.基因转移是将外源DNA或基因序列导入 转化为宿主体细胞，这种方法有助于体内 研究基因表达和功能。重组体的递送 基因进入肺血管可能会导致 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 细胞这些细胞中的基因表达可以促进 肺血管疾病的病理生物学研究和 基因治疗方法的潜在发展，以治疗这些血管 疾病这项补助金的目的是检验假设， 重组基因可以在肺内递送和表达， 体内的脉管系统。在特定目标I中，将重组报告基因 使用腺病毒和DNA导入肺血管 脂质体载体，基因表达将通过DNA分析确认， mRNA和蛋白质;将定义转导的细胞类型;以及 肺和其他组织中基因表达的持续时间将是 研究了在具体目标II中，将对这些内容进行修改。 载体以优化体内转染效率和基因表达。 将使用表面活性剂开发肺特异性基因表达 发起人。在《特定目的III》中，重组基因可能 与成人组织相比， 通过研究以下基因表达的稳定性进行测试： 用腺病毒和DNA脂质体载体将基因转移到新生儿中， 成人肺我们实验室的初步研究表明， 这些研究的可行性。在猪的动物模型中，我们已经证明 重组报告基因可以在细胞中的特定位点表达， 经导管注射后的肺血管。在人类基因中 治疗I期研究，经导管递送HLA-B7 DNA和脂质体 右后基底段动脉是安全的， 黑色素瘤转移到右下叶的患者。这个目标 建议是优化重组基因的递送和表达 在直接基因转移后的肺血管中， 腺病毒和DNA脂质体载体。此外，发展 将研究新生儿和成人中基因表达的差异 肺早期表达这些重组基因的能力可能 允许早期干预，以防止疾病的进展， 肺部疾病。综合起来看，这些研究应该能提供 肺组织内基因表达的基本机制， 为肺血管性疾病的基因治疗提供了一条途径 疾病

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.