New York University School of Medicine, Hansjörg Wyss Department of Plastic and Reconstructive Surgery, 430 East 29th Street, New York, NY, 10016, United States.
New York University Tandon School of Engineering, Chemical and Biomolecular Engineering Department, 6 Metrotech Center, Brooklyn, NY 11201, United States.
Biomaterials. 2017 Jul;132:1-15. doi: 10.1016/j.biomaterials.2017.04.001. Epub 2017 Apr 3.
Therapeutics utilizing siRNA are currently limited by the availability of safe and effective delivery systems. Cutaneous diseases, specifically ones with significant genetic components are ideal candidates for topical siRNA based therapy but the anatomical structure of skin presents a considerable hurdle. Here, we optimized a novel liposome and protein hybrid nanoparticle delivery system for the topical treatment of diabetic wounds with severe oxidative stress. We utilized a cationic lipid nanoparticle (CLN) composed of 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) and the edge activator sodium cholate (NaChol), in a 6:1 ratio of DOTAP:NaChol (DNC). Addition of a cationic engineered supercharged coiled-coil protein (CSP) in a 10:1:1 ratio of DNC:CSP:siRNA produced a stable lipoproteoplex (LPP) nanoparticle, with optimal siRNA complexation, minimal cytotoxicity, and increased transfection efficacy. In a humanized murine diabetic wound healing model, our optimized LPP formulation successfully delivered siRNA targeted against Keap1, key repressor of Nrf2 which is a central regulator of redox mechanisms. Application of LPP complexing siKeap1 restored Nrf2 antioxidant function, accelerated diabetic tissue regeneration, and augmented reduction-oxidation homeostasis in the wound environment. Our topical LPP delivery system can readily be translated into clinical use for the treatment of diabetic wounds and can be extended to other cutaneous diseases with genetic components.
目前，利用 siRNA 的治疗方法受到安全有效的输送系统的限制。皮肤疾病，特别是具有重要遗传成分的疾病，是局部 siRNA 治疗的理想候选者，但皮肤的解剖结构构成了相当大的障碍。在这里，我们优化了一种新型的脂质体和蛋白质混合纳米颗粒输送系统，用于治疗严重氧化应激的糖尿病伤口。我们利用了一种由 1,2-二油酰基-3-三甲铵丙烷（DOTAP）和边缘激活剂脱氧胆酸钠（NaChol）组成的阳离子脂质体（CLN），DOTAP:NaChol 的比例为 6:1（DNC）。在 DNC:CSP:siRNA 的 10:1:1 比例下添加阳离子工程超荷卷曲螺旋蛋白（CSP），产生了稳定的脂蛋白复合物（LPP）纳米颗粒，具有最佳的 siRNA 复合物形成、最小的细胞毒性和增强的转染效率。在人源化的糖尿病小鼠伤口愈合模型中，我们优化的 LPP 配方成功地递送了针对 Keap1 的 siRNA，Keap1 是 Nrf2 的关键抑制剂，Nrf2 是氧化还原机制的中央调节剂。应用 LPP 复合物的 siKeap1 恢复了 Nrf2 的抗氧化功能，加速了糖尿病组织的再生，并增加了伤口环境中的氧化还原平衡。我们的局部 LPP 输送系统可以很容易地转化为治疗糖尿病伤口的临床应用，并可以扩展到其他具有遗传成分的皮肤疾病。
