CATIONIC LIPIDS FOR CFTR GENE THERAPY

我们实验室已经积累了基于基因的阳离子脂质体经验 调职。我们已经将这一方法应用于临床前的CFTR交付 体外培养人呼吸道上皮细胞的研究 活体哺乳动物呼吸道上皮细胞。此外，临床前研究 利用阳离子脂质体进行基因转移的研究已经迅速发展起来 使用DMRIE/Dope载体进行CFTR转移的人体临床试验。 这些基础和临床前的发展已经产生了对正式的 启动涉及慢性萎缩性胃炎患者的治疗和临床试验的手段。 该项目旨在提供必要的资源，以开发和 完成9例CFTR基因鼻腔内转移的临床试验 使用阳离子脂质体的患者。这项研究是迈向 基于脂质体的CFTR治疗的未来试验，包括对 反复鼻腔给药，下呼吸道给药 Cf患者Iipid/DNA偶联物及其他阳离子脂类的研究 或体内人类基因给药过程中的脂质/DNA比率。我们会 进行必要的临床分析，以促进未来对CF的人体研究 在UAB中心。这些化验包括：1)鼻部电位的测量 2)RT-PCR检测正常CFTRm RNA跟随基因 3)体内原代培养的呼吸道上皮细胞 基因转移；4)CFTR突变分析。我们还将研究 脂基基因的基因传递和表达机制 体外和体内转移。制定的原则和方法 将为今后的研究奠定坚实的基础 脂质体CFTR基因治疗。

Our laboratory has developed experience with cationic Liposome based gene transfer. We have applied this methodology to CFTR delivery in preclinical studies to human respiratory epithelial cells in vitro, as well as to mammalian respiratory epithelium in vivo. In addition, preclinical studies of gene transfer using cationic liposomes have rapidly led to development of a human clinical trial of CFTR transfer using a DMRIE/DOPE vehicle. These basic and preclinical developments have Created the need for a formal means for initiating therapeutic, clinical trials involving CF patients. This project intends to provide the resources necessary to develop and complete a clinical trial of CFTR gene transfer to the nasal airway in 9 CF patients using cationic Iiposomes. This study is the first step towards future trials of liposome based CFTR therapy, including evaluation of repeated nasal administration of CFTR, lower airway administration of Iipid/DNA conjugates to CF patients, and studies of other cationic lipids or lipid/DNA ratios during in vivo human gene administration. We will perform clinical assays necessary to facilitate future human studies of CF at the UAB center. These assays include 1) measurement of nasal potential difference; 2) RT-PCR detection of normal CFTR mRNA following gene transfer; 3) primary culture of airway epithelial cells following in vivo gene transfer; and 4) CFTR mutation analysis. We will also examine the mechanisms of gene delivery and expression underlying lipid based gene transfer in vitro and in vivo. The principles and approaches developed within this project will form a solid foundation for future studies of liposomal CFTR gene therapy.