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CFTR Delivery to Ciliated Airway Cells by PIV Vectors

通过 PIV 载体将 CFTR 递送至纤毛气道细胞

基本信息

批准号：
7252023
负责人：
RAYMOND J PICKLES
金额：
$ 34.45万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-09-06 至 2010-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7252023
关键词：
Accounting Apical Appendix Attenuated Bacterial Infections Biology Cells Chronic Clinical Trials Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Cytopathology Dehydration Development Disease Dose Epithelial Cells Epithelium Gene Delivery Gene Transfer Generations Genes Genome Homeostasis Human Immune response Immune system In Vitro Infection Liquid substance Lung Modeling Morbidity - disease rate Nasal Epithelium Nose Numbers Phenotype Physiological Proteins Pulmonary Cystic Fibrosis Recombinants Respiratory physiology Respiratory syncytial virus Rodent Safety Structure of respiratory epithelium Surface Testing Transgenes Viral Viral Genome Virus airway epithelium base cell type concept cystic fibrosis airway epithelia cystic fibrosis patients cytotoxic cytotoxicity gene transfer vector immunogenic immunogenicity in vitro Model in vivo in vivo Model knowledge base mortality nonhuman primate novel parainfluenza virus prototype respiratory virus vector

项目摘要

DESCRIPTION (provided by applicant): Cystic fibrosis (CF) lung disease causes dehydration of the surface lining of the respiratory epithelium leading to mucostasis and chronic bacterial infection. The absent gene product in CF is the cystic fibrosis transmembrane conductance regulator (CFTR) protein that regulates fluid homeostatic mechanisms in the airway epithelium. We hypothesize that expression of functional CFTR in the airway epithelium of CF patients will restore CFTR function and thus normal lung function. Gene transfer strategies for CF lung disease have so far failed to show significant CFTR delivery to airway cells in vivo, primarily because the vectors did not efficiently transfer CFTR to the respiratory epithelium after intraluminal delivery. We have found that recombinant parainfluenza viruses (rPIV), infect cultures of human well-differentiated airway cells (HAE) with high efficiency after intraluminal delivery, and that gene transfer is specific to ciliated airway epithelial cells, the cell type requiring correction in CF. The high efficiency of gene transfer by rPIV to ciliated cells, the availability of recombinant versions of PIV and, the capacity to insert large transgenes into the rPIV genome suggests that these viruses may be useful for delivering CFTR to the ciliated airway epithelium of the CF lung. However, currently available PIV vectors are cytotoxic to ciliated cells. Therefore, we propose to generate novel recombinant rPIV-based gene transfer vectors that express human CFTR and test the efficacy and safety of these vectors at correcting the CF phenotype of CF HAE. We will also test the efficency, safety and the influence of the immune system on gene transfer by rPIV-vectors in the nasal epithelium of non-human primates in vivo. The Specific Aims of the proposed study are: 1) Can rPIV-Vectors be Generated for Efficacious and Safe CFTR gene transfer to Ciliated Airway Cells? 2) To Test the Efficacy and Cytotoxicity of rPIV-CFTR vectors on Human Well-Differentiated Airway Epithelial Cells In Vitro; and, 3) To Test the Gene Transfer Efficiency of rPIV-Vectors in the Non-Human Primate Nasal Epithelium In Vivo.

描述(由申请人提供)：囊性纤维化(CF)肺部疾病导致呼吸道上皮表面衬里脱水，导致粘液淤滞和慢性细菌感染。在CF中缺失的基因产物是囊性纤维化跨膜传导调节蛋白(CFTR)，它调节呼吸道上皮细胞的液体稳态机制。我们假设，在CF患者的呼吸道上皮细胞中表达功能性CFTR将恢复CFTR功能，从而恢复正常的肺功能。到目前为止，治疗慢性肺病的基因转移策略还没有显示出显著的CFTR体内递送到呼吸道细胞，主要是因为载体在腔内递送后没有有效地将CFTR转移到呼吸道上皮细胞。我们发现，重组副流感病毒(RPIV)在腔内输送后高效地感染人高分化的呼吸道细胞(HAE)，并且基因转移是针对纤毛呼吸道上皮细胞的，这是CF中需要纠正的细胞类型。RPIV基因转移到纤毛细胞的高效率，重组PIV的可获得性，以及将大量转基因插入rPIV基因组的能力，表明这些病毒可能有助于将CFTR转移到CF肺的纤毛呼吸道上皮。然而，目前可用的PIV载体对纤毛细胞具有细胞毒性。因此，我们建议构建新型的重组rPIV基因转移载体来表达人CFTR，并测试这些载体在纠正CFHAE的CF型方面的有效性和安全性。我们还将在体内测试免疫系统对rPIV载体在非人类灵长类动物鼻黏膜上皮细胞中基因转移的有效性、安全性和影响。本研究的具体目的是：1)能否产生rPIV-Vectors，用于将CFTR基因转移到纤毛细胞？2)检测rPIV-CFTR载体对体外培养的人高分化呼吸道上皮细胞的有效性和细胞毒作用；3)检测rPIV-Vectors在体内非人灵长类鼻黏膜上皮细胞中的基因转移效率。