Fargnoli Anthony S, Mu Anbin, Katz Michael G, Williams Richard D, Margulies Kenneth B, Weiner David B, Yang Shu, Bridges Charles R
Thoracic and Cardiovascular Surgery, Sanger Heart & Vascular Institute, Carolinas Healthcare System, Charlotte, NC, USA.
J Transl Med. 2014 Jun 16;12:171. doi: 10.1186/1479-5876-12-171.
Cardiac gene therapy for heart disease is a major translational research area with potential, yet problems with safe and efficient gene transfer into cardiac muscle remain unresolved. Existing methodology to increase vector uptake include modifying the viral vector, non-viral particle encapsulation and or delivery with device systems. These advanced methods have made improvements, however fail to address the key problem of inflammation in the myocardium, which is known to reduce vector uptake and contribute to immunogenic adverse events. Here we propose an alternative method to co-deliver anti-inflammatory drugs in a controlled release polymer with gene product to improve therapeutic effects.
A robust, double emulsion production process was developed to encapsulate drugs into nanoparticles. Briefly in this proof of concept study, aspirin and prednisolone anti-inflammatory drugs were encapsulated in various poly-lactic glycolic acid polymer (PLGA) formulations. The resultant particle systems were characterized, co-delivered with GFP plasmid, and evaluated in harvested myocytes in culture for uptake.
High quality nanoparticles were harvested from multiple production runs, with an average 64 ± 10 mg yield. Four distinct particle drug system combinations were characterized and evaluated in vitro: PLGA(50:50) Aspirin, PLGA(65:35) Prednisolone, PLGA(65:35) Aspirin and PLGA(50:50) Prednisolone Particles consisted of spherical shape with a narrow size distribution 265 ± 104 nm as found in scanning electron microscopy imaging. Prednisolone particles regardless of PLGA type were found on average ≈ 100 nm smaller than the aspirin types. All four groups demonstrated high zeta potential stability and re-constitution testing prior to in vitro. In vitro results demonstrated co uptake of GFP plasmid (green) and drug loaded particles (red) in culture with no incidence of toxicity.
Nano formulated anti-inflammatories in combination with standalone gene product therapy may offer a clinical solution to maximize cardiac gene therapy product effects while minimizing the risk of the host response in the inflammatory myocardial environment.
用于治疗心脏病的心脏基因疗法是一个具有潜力的重要转化研究领域，但将基因安全有效地导入心肌的问题仍未得到解决。现有的增加载体摄取的方法包括修饰病毒载体、非病毒颗粒封装和/或使用装置系统进行递送。这些先进方法虽有改进，但未能解决心肌炎症这一关键问题，已知炎症会降低载体摄取并导致免疫原性不良事件。在此，我们提出一种替代方法，即将抗炎药物与基因产物共同包裹于控释聚合物中以提高治疗效果。
开发了一种稳健的双乳液生产工艺，将药物封装到纳米颗粒中。在此概念验证研究中，简要来说，阿司匹林和泼尼松龙这两种抗炎药物被封装于各种聚乳酸 - 乙醇酸聚合物（PLGA）制剂中。对所得颗粒系统进行表征，与绿色荧光蛋白（GFP）质粒共同递送，并在培养的收获心肌细胞中评估摄取情况。
通过多次生产运行收获了高质量的纳米颗粒，平均产量为64±10毫克。对四种不同的颗粒药物系统组合进行了表征并在体外进行评估：PLGA（50:50）阿司匹林、PLGA（65:35）泼尼松龙、PLGA（65:35）阿司匹林和PLGA（50:50）泼尼松龙。颗粒呈球形，扫描电子显微镜成像显示尺寸分布狭窄，为265±104纳米。无论PLGA类型如何，泼尼松龙颗粒平均比阿司匹林颗粒小约100纳米。所有四组在体外之前均表现出高zeta电位稳定性和重构测试。体外结果表明，培养物中绿色荧光蛋白质粒（绿色）和载药颗粒（红色）共同摄取，且无毒性发生。
纳米配方的抗炎药与单独的基因产物疗法相结合，可能提供一种临床解决方案，在炎症性心肌环境中最大化心脏基因治疗产品的效果，同时将宿主反应的风险降至最低。
