GENE TRANSFER INTO THE PULMONARY VASCULATURE

许多心肺疾病的研究和治疗受到以下限制 在非分裂细胞中表达特定重组基因的能力 活着。基因转移是指将外源DNA或基因序列导入 进入宿主体细胞，这种方法便于在体内 基因表达和功能的研究。重组人的交付 进入肺血管系统的基因可能导致 重组基因在大量细胞中的表达，包括 内皮细胞、II型肺泡细胞和远端呼吸道上皮细胞 细胞。基因在这些细胞中的表达可以促进 肺血管疾病的病理生物学研究及临床意义 治疗这些血管的基因治疗方法的潜在发展 疾病。这笔拨款的目的是检验这一假设 重组基因可在肺组织中传递和表达 体内的血管构筑。在特定的目标I中，重组报告基因将被 用腺病毒和DNA导入肺血管系统 脂质体载体，基因表达将通过DNA分析得到确认， M RNA和蛋白质；被转导的细胞类型；以及 基因在肺和其他组织中表达的持续时间将是 调查过了。在具体目标二中，将对这些内容进行修改 载体，以优化体内的转染率和基因表达。 将使用表面活性物质开发肺特异基因表达 推动者。在特定目标III中，重组基因可能 与成人组织相比，新生儿组织中的存留时间更长 通过对以下基因表达稳定性的调查进行测试 腺病毒和DNA脂质体载体转导新生大鼠的研究 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，我们已经展示了 重组报告基因可以在细胞中的特定位置表达。 经导管注射后的肺血管系统。在人类基因中 治疗I期研究，经导管输送人类白细胞抗原-B7 DNA和脂质体 进入右后基底段动脉的手术是安全的 1例黑色素瘤转移至右下叶。这样做的目的是 建议优化重组基因的传递和表达 直接基因转移后的肺血管内皮细胞 腺病毒和DNA脂质体载体。此外，发展中的 新生儿和成人的基因表达差异将被调查。 肺部。在早期表达这些重组基因的能力可能会 允许进行早期干预，旨在防止疾病的进展 肺部疾病。综上所述，这些研究应该会提供洞察力 研究肺组织中基因表达的基本机制，并可能 为肺血管疾病的基因治疗提供一条途径 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.