Pseudotyping FIV to Target Human Airway Epithelia

描述（申请人？摘要）：目前存在的几个问题限制了 基因治疗在囊性纤维化肺病治疗中的应用。 大多数载体系统的一个显著限制是， 当它们应用于气道上皮细胞的顶端表面时，基因转移。 一个有吸引力的方法来克服这个问题涉及替代 （假型化）整合猫的包膜糖蛋白 基于免疫缺陷病毒（FIV）的慢病毒载体与病毒载体 其可能靶向气道上皮细胞顶端表面上的受体。与 最后，我们从多个病毒家族中筛选了几种包膜蛋白， 包括嗜热性、异嗜性、VSV-G、RD 114、IOA 1、GALV、马尔堡和 埃博拉病毒包膜糖蛋白。除丝状病毒外的所有假型 家族在从顶端表面转导细胞方面是无效的。然而，在这方面， 初步结果表明，基于FIV的载体与马尔堡 包膜糖蛋白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， 逆转录病毒载体对基因转移的绝对顶端屏障。我们提出 评价马尔堡假型FIV载体有效性的研究 以促进基因转移穿过气道上皮的顶端表面。在 此外，我们将测试此伪类型纠正CF的能力 通过用表达CFTR的病毒载体感染，在CF上皮细胞中观察CFTR表型。我们 进一步提出技术-以提高滴度以及证明在 这种新型载体的体内功效。成功完成这些建议 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.