Mahajan Vivek, Gaymalov Zagit, Alakhova Daria, Gupta Richa, Zucker Irving H, Kabanov Alexander V
Division of Molecular Pharmaceutics, Center for Nanotechnology in Drug Delivery, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, NC 27599, USA; Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, NE 68198-5850, USA.
Division of Molecular Pharmaceutics, Center for Nanotechnology in Drug Delivery, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, NC 27599, USA; Department of Pharmaceutical Sciences and Center for Drug Delivery and Nanomedicine, College of Pharmacy, University of Nebraska Medical Center, Omaha, NE 68198-5850, USA.
Biomaterials. 2016 Jan;75:58-70. doi: 10.1016/j.biomaterials.2015.10.002. Epub 2015 Oct 9.
Intramuscular administration of plasmid DNA (pDNA) with non-ionic Pluronic block copolymers increases gene expression in injected muscles and lymphoid organs. We studied the role of immune cells in muscle transfection upon inflammation. Local inflammation in murine hind limb ischemia model (MHLIM) drastically increased DNA, RNA and expressed protein levels in ischemic muscles injected with pDNA/Pluronic. The systemic inflammation (MHLIM or peritonitis) also increased expression of pDNA/Pluronic in the muscles. When pDNA/Pluronic was injected in ischemic muscles the reporter gene, Green Fluorescent Protein (GFP) co-localized with desmin(+) muscle fibers and CD11b(+) macrophages (MØs), suggesting transfection of MØs along with the muscle cells. P85 enhanced (∼ 4 orders) transfection of MØs with pDNA in vitro. Moreover, adoptively transferred MØs were shown to pass the transgene to inflamed muscle cells in MHLIM. Using a co-culture of myotubes (MTs) and transfected MØs expressing a reporter gene under constitutive (cmv-luciferase) or muscle specific (desmin-luciferase) promoter we demonstrated that P85 enhances horizontal gene transfer from MØ to MTs. Therefore, MØs can play an important role in muscle transfection with pDNA/Pluronic during inflammation, with both inflammation and Pluronic contributing to the increased gene expression. pDNA/Pluronic has potential for therapeutic gene delivery in muscle pathologies that involve inflammation.
将质粒DNA（pDNA）与非离子型普朗尼克嵌段共聚物进行肌肉注射，可增加注射肌肉和淋巴器官中的基因表达。我们研究了免疫细胞在炎症时肌肉转染中的作用。小鼠后肢缺血模型（MHLIM）中的局部炎症显著增加了注射pDNA/普朗尼克的缺血肌肉中的DNA、RNA和表达蛋白水平。全身炎症（MHLIM或腹膜炎）也增加了肌肉中pDNA/普朗尼克的表达。当将pDNA/普朗尼克注射到缺血肌肉中时，报告基因绿色荧光蛋白（GFP）与结蛋白（+）肌纤维和CD11b（+）巨噬细胞（MØs）共定位，提示MØs与肌肉细胞一起被转染。P85在体外增强了（约4个数量级）pDNA对MØs的转染。此外，在MHLIM中，过继转移的MØs被证明可将转基因传递给炎症肌肉细胞。利用在组成型（cmv-荧光素酶）或肌肉特异性（结蛋白-荧光素酶）启动子下表达报告基因的肌管（MTs）和转染MØs的共培养，我们证明P85增强了从MØ到MTs的水平基因转移。因此，在炎症期间，MØs在pDNA/普朗尼克介导的肌肉转染中可发挥重要作用，炎症和普朗尼克均有助于增加基因表达。pDNA/普朗尼克在涉及炎症的肌肉疾病中具有治疗性基因递送的潜力。
