Targeting Vector Interactome to Enhance CFTR Delivery

摘要 DNA纳米颗粒已经在CF患者中进行了测试，并且显示出是安全的，并且能够将核酸递送到CF患者。 气道。我们已经报道了核仁素与糖皮质激素受体（GR）的相互作用是必不可少的， 细胞结合和转染，并发现DNPs与人类中的其他蛋白质相互作用， 原代气道上皮细胞，包括腺瘤性结肠息肉病（APC）和非红细胞血影蛋白 α 1（SPTAN1）。这些相互作用可以显著调节DNP介导的基因转移，这促使 我们假设DNP与GR、HSP 90、APC和SPTAN 1的相互作用调节核仁素介导的 颗粒的运输，并且可以用在人体中批准的试剂操纵相互作用， 增强体内基因递送。为了研究这一假设，我们的具体目标是：1）评估影响 操纵DNP相互作用组对体内基因递送和毒性的影响;和2）研究DNP相互作用组对体内基因递送和毒性的作用机制。 其增强来自CF患者的分化良好的气道上皮细胞中的基因转移。拟议的研究 将是第一个探索DNP生物学并提高我们对交付重要步骤的理解。 此外，这些研究有可能将已经批准用于人类的小分子定义为 可以在纳米颗粒给药之前或期间给药以显著增强基因表达的试剂 将CFTR转移到治疗范围内。

Abstract DNA nanoparticles have been tested in CF patients and shown to be safe and able to deliver nucleic acids to the airway. We have reported that nucleolin interaction with glucocorticoid receptor (GR) is essential for cellular binding and transfection of DNPs and discovered that DNPs interact with other proteins in human primary airway epithelial cells, including adenomatous polyposis coli (APC), and non-erythrocytic spectrin alpha 1 (SPTAN1). These interactions can significantly modulate DNPs-mediated gene transfer, which prompts our hypothesis that DNP interaction with GR, HSP90, APC, and SPTAN1 modulate nucleolin-mediated transport of the particles and that the interactions can be manipulated with agents approved in humans to enhance gene delivery in vivo. To investigate the hypothesis, our specific aims are: 1) To evaluate the impact of manipulating the DNP interactome on gene delivery and toxicity in vivo; and 2) To examine the mechanisms that enhance gene transfer in well differentiated airway epithelial cells from CF patients. The proposed studies will be the first to explore the biology of DNPs and improve our understanding of important steps in delivery. Furthermore, the studies have the potential of defining small molecules already approved for use in humans as agents that can be administered before or during nanoparticle administration to significantly enhance gene transfer of CFTR into the therapeutic range.