College of Pharmacy and Nutrition, University of Saskatchewan, Saskatoon, Saskatchewan, Canada.
Gene Ther. 2012 Oct;19(10):978-87. doi: 10.1038/gt.2011.159. Epub 2011 Nov 10.
Scleroderma is a chronic disorder manifested by excessive synthesis and deposition of collagen in skin and connective tissue, vascular abnormalities, and autoimmunity. Using microarray and real-time PCR data, we show that intradermally expressed interferon γ (IFN-γ), generated after intradermal injection of IFN-γ-coding plasmid, and non-invasive topical nanoparticle (TNP) treatment with IFN-γ-coding plasmid, decreased collagen synthesis (via the Jak/Stat 1 pathway), upregulated Th1 cytokine levels, and downregulated the profibrotic cytokine Transforming growth factor β and the Smad pathways in the Tsk/+ (tight-skin scleroderma) mouse model. The TNP gene delivery system was constructed from gemini surfactant 16-3-16 and IFN-γ-coding plasmid. Topical administration of IFN-γ-coding plasmid in TNPs was effective in expressing IFN-γ levels after a 20-day treatment regimen without increased TLR4, CCL2, CCL11 and CCR2 mRNA levels that were observed in injected animals, signs considered to be innate responses to injury. The more uniform transgene IFN-γ expression caused significant (70-72%) collagen reduction, as assessed by reverse transcription real-time PCR. These results demonstrate efficient in vivo transfection using a gemini surfactant-based TNP delivery system able to modulate excessive collagen synthesis in scleroderma-affected skin.
硬皮病是一种慢性疾病，表现为皮肤和结缔组织中胶原过度合成和沉积、血管异常和自身免疫。我们使用微阵列和实时 PCR 数据表明，经皮注射 IFN-γ 编码质粒后产生的真皮内表达的干扰素 γ（IFN-γ）和非侵入性局部纳米颗粒（TNP）治疗 IFN-γ 编码质粒，可减少胶原蛋白合成（通过 Jak/Stat 1 途径），上调 Th1 细胞因子水平，并下调 Tsk/+（紧皮硬皮病）小鼠模型中的促纤维化细胞因子转化生长因子 β 和 Smad 途径。TNP 基因传递系统由双子表面活性剂 16-3-16 和 IFN-γ 编码质粒构建而成。在 TNP 中局部给予 IFN-γ 编码质粒，在 20 天的治疗方案后有效表达 IFN-γ 水平，而在注射动物中观察到的 TLR4、CCL2、CCL11 和 CCR2 mRNA 水平没有增加，这些被认为是对损伤的先天反应的迹象。更均匀的转基因 IFN-γ 表达导致逆转录实时 PCR 评估的胶原蛋白减少 70-72%。这些结果表明，使用基于双子表面活性剂的 TNP 传递系统能够有效转染体内，能够调节硬皮病皮肤中过度的胶原蛋白合成。
