Targeting Vector Interactome to Enhance CFTR Delivery

摘要 DNA纳米颗粒已经在CF患者身上进行了测试，并被证明是安全的，能够将核酸输送到 呼吸道。我们已经报道，核仁素与糖皮质激素受体(GR)的相互作用是 DNPs的细胞结合和转染，并发现DNPs与人体内其他蛋白质相互作用 原代呼吸道上皮细胞，包括结肠腺瘤性息肉病(APC)和非红细胞性血影 Alpha 1(SPTAN1)。这些相互作用可以显著调节DNPs介导的基因转移，这促使 我们的假设是DNP与GR、HSP90、APC和SPTAN1的相互作用调节核仁素介导的 粒子的传输和相互作用可以通过在人体内批准的试剂来操纵 增强体内的基因传递。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传递和毒性的影响；以及2)研究其机制 增强CF患者分化良好的呼吸道上皮细胞的基因转移。建议进行的研究 将第一个探索DNPs的生物学并提高我们对交付过程中重要步骤的理解。 此外，这些研究有可能将已经被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的药物 将CFTR转移到治疗范围。

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.