MacLoughlin Ronan J, Higgins Brendan D, Devaney James, O'Toole Daniel, Laffey John G, O'Brien Timothy
1 Regenerative Medicine Institute (REMEDI), National University of Ireland , Galway, Ireland .
Hum Gene Ther. 2015 Jan;26(1):36-46. doi: 10.1089/hum.2014.053.
Inhibition of the proinflammatory transcription factor NF-κB has previously been shown to attenuate the inflammatory response in tissue after injury. However, the feasibility and efficacy of aerosolized adeno-associated viral (AAV) vector-delivered transgenes to inhibit the NF-κB pathway are less clear. Initial studies optimized the AAV vector for delivery of transgenes to the pulmonary epithelium. The effect of repeated nebulization on the integrity and transduction efficacy of the AAV vector was then examined. Subsequent in vivo studies examined the efficacy of aerosolized rAAV2/6 overexpressing the NF-κB inhibitor IκBα in a rodent endotoxin-induced lung injury model. Initial in vitro investigations indicated that rAAV2/6 was the most effective vector to transduce the lung epithelium, and maintained its integrity and transduction efficacy after repeated nebulization. In our in vivo studies, animals that received aerosolized rAAV2/6-IκBα demonstrated a significant increase in total IκBα levels in lung tissue relative to null vector-treated animals. Aerosolized rAAV2/6-IκBα attenuated endotoxin-induced bronchoalveolar lavage-detected neutrophilia, interleukin-6 and cytokine-induced neutrophil chemoattractant-1 levels, as well as total protein content, and decreased histologic indices of injury. These results demonstrate that aerosolized AAV vectors encoding human IκBα significantly attenuate endotoxin-mediated lung injury and may be a potential therapeutic candidate in the treatment of acute lung injury.
先前的研究表明,抑制促炎转录因子NF-κB可减轻组织损伤后的炎症反应。然而,雾化腺相关病毒(AAV)载体递送转基因以抑制NF-κB途径的可行性和有效性尚不清楚。初步研究优化了用于将转基因递送至肺上皮的AAV载体。然后研究了重复雾化对AAV载体完整性和转导效率的影响。随后的体内研究在啮齿动物内毒素诱导的肺损伤模型中检测了雾化表达NF-κB抑制剂IκBα的重组腺相关病毒2/6(rAAV2/6)的疗效。最初的体外研究表明,rAAV2/6是转导肺上皮最有效的载体,并且在重复雾化后保持其完整性和转导效率。在我们的体内研究中,接受雾化rAAV2/6-IκBα的动物相对于空载体处理的动物,肺组织中总IκBα水平显著增加。雾化rAAV2/6-IκBα减轻了内毒素诱导的支气管肺泡灌洗检测到的中性粒细胞增多、白细胞介素-6和细胞因子诱导的中性粒细胞趋化因子-1水平以及总蛋白含量,并降低了组织学损伤指标。这些结果表明,雾化编码人IκBα的AAV载体可显著减轻内毒素介导的肺损伤,可能是治疗急性肺损伤的潜在候选疗法。