Pseudotyping FIV to Target Human Airway Epithelia

描述(申请人？S摘要)：目前有几个问题限制了 基因治疗在囊性纤维性肺病治疗中的应用。 对于大多数载体系统来说，一个重要的限制是 当它们应用于呼吸道上皮细胞的顶面时，基因转移。 解决这一问题的一个有吸引力的方法是用 (伪分型)整合猫科动物的包膜糖蛋白 基于免疫缺陷病毒(FIV)的慢病毒载体与病毒载体 这可能以呼吸道上皮细胞顶端表面的受体为靶点。到那时候 最后，我们已经从多个病毒家族中筛选出几个包膜蛋白， 包括两性、异向性、VSV-G、RD 114、IOA1、Galv、Marburg和 埃博拉病毒包膜糖蛋白。除丝状病毒外的所有伪型 家系在从根尖表面转导细胞方面无效。然而， 初步结果表明，基于FIV的载体与Marburg 包膜糖蛋白应用于皮肤上皮细胞时，具有感染上皮细胞的能力 顶面。这些结果令人鼓舞，因为它们表明没有 用逆转录病毒载体进行基因转移的绝对顶端屏障。我们建议 将评估马尔堡伪型FIV载体效率的研究 以促进跨越呼吸道上皮细胞顶面的基因转移。在……里面 此外，我们将测试此伪型纠正CF的能力 表达CFTR的病毒载体感染CF上皮细胞的表型我们 进一步提出技术-以提高滴度并展示 这种新型载体的体内效力。圆满完成这些拟议的 实验对基因治疗的发展具有重要意义。

DESCRIPTION (Applicant?s Abstract): Several problems currently limit the application of gene therapy for the treatment for cystic fibrosis lung disease. One significant limitation for most vector systems is the poor efficiency of gene transfer when they are applied to the apical surface of airway epithelia. An attractive approach for overcoming this problem involves substituting (pseudotyping) the envelope glycoproteins of the integrating Feline Immunodeficiency Virus (FIV)-based lentivirus vectors with those of viruses that may target receptors on the apical surface of airway epithelia. To that end, we have screened several envelope proteins from multiple viral families, including amphotropic, xenotropic, VSV-G, RD 114, I OA 1, GALV, Marburg, and Ebola envelope glycoproteins. All pseudotypes except those from the filovirus family were ineffective in transducing cells from the apical surface. However, initial results suggest that the FIV-based vector pseudotyped with the Marburg envelope glycoprotein has the capacity to infect epithelia when applied to the apical surface. These results are encouraging as they suggest that there is no absolute apical barrier to gene transfer with retrovirus vectors. We propose studies that will evaluate the efficiency of the Marburg pseudotyped FIV vector to promote gene transfer across the apical surface of airway epithelia. In addition, we will test-the ability of this pseudotype to correct the CF phenotype in CF epithelia by infecting with a CFTR expressing viral vector. We further propose techniques -to improve the titer as well as demonstrate the in vivo efficacy of this novel vector. The successful completion of these proposed experiments has great implications for the advancement gene therapy.