title: Imaging Dendrimer-Grafted Graphene Oxide Mediated Anti-miR-21 Delivery With an Activatable Luciferase Reporter.
abstract: MicroRNAs (miRNAs) are a class of post-transcriptional gene regulators involved in various physiological processes including carcinogenesis, and they have emerged as potential targets for tumor theranostics. However, the employment of antisense oligonucleotides, termed anti-miRs, for antagonizing miRNA functions in vivo has largely been impeded by a lack of effective delivery carriers. Here, we describe the development of polyamidoamine (PAMAM) dendrimer and polyethylene glycol (PEG)-functionalized nanographene oxide (NGO) conjugate (NGO-PEG-dendrimer) for the efficient delivery of anti-miR-21 into non-small-cell lung cancer cells. To monitor the delivery of anti-miR-21 into cells and tumors, we also constructed an activatable luciferase reporter (Fluc-3xPS) containing three perfectly complementary sequences against miR-21 in the 3' untranslated region (UTR) of the reporter. Compared with bare dendrimer and Lipofectamine 2000 (Lipo2000), NGO-PEG-dendrimer showed considerably lower cytotoxicity and higher transfection efficiency. As demonstrated by in vitro bioluminescence imaging and Western blotting assays, NGO-PEG-dendrimer effectively delivered anti-miR-21 into the cytoplasm and resulted in the upregulation of luciferase intensity and PTEN target protein expression in a dose-dependent manner. Moreover, transfection with anti-miR-21 by NGO-PEG-dendrimer led to stronger inhibition of cell migration and invasion than did bare dendrimer or Lipo2000 transfection. The intravenous delivery of anti-miR-21 via NGO-PEG-dendrimer induced a significant increase in the bioluminescence signal within the Fluc-3xPS reporter-transplanted tumor areas. These results suggest that NGO-PEG-dendrimer could be an efficient and a potential nanocarrier for delivering RNA oligonucleotides. In addition, the strategy of combining NGO-PEG-dendrimer with an activatable luciferase reporter allows the image-guided monitoring of the delivery process, which can provide insights into the RNA-based cancer treatments.
authors.affiliation: Engineering Research Center of Molecular and Neuro Imaging, Ministry of Education, School of Life Science and Technology, Xidian University , Xi'an, Shaanxi 710071, China.
authors.forename: Fu
authors.initials: F
authors.lastname: Wang
authors.affiliation: Department of Gynecology and Obstetrics, Tangdu Hospital, The Fourth Military Medical University, Xi'an , Shaanxi 710038, China.
authors.forename: Beilei
authors.initials: B
authors.lastname: Zhang
authors.affiliation: Engineering Research Center of Molecular and Neuro Imaging, Ministry of Education, School of Life Science and Technology, Xidian University , Xi'an, Shaanxi 710071, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Zhou
authors.affiliation: Engineering Research Center of Molecular and Neuro Imaging, Ministry of Education, School of Life Science and Technology, Xidian University , Xi'an, Shaanxi 710071, China.
authors.forename: Yaru
authors.initials: Y
authors.lastname: Shi
authors.affiliation: Engineering Research Center of Molecular and Neuro Imaging, Ministry of Education, School of Life Science and Technology, Xidian University , Xi'an, Shaanxi 710071, China.
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Li
authors.affiliation: Engineering Research Center of Molecular and Neuro Imaging, Ministry of Education, School of Life Science and Technology, Xidian University , Xi'an, Shaanxi 710071, China.
authors.forename: Yuqiong
authors.initials: Y
authors.lastname: Xia
authors.affiliation: Engineering Research Center of Molecular and Neuro Imaging, Ministry of Education, School of Life Science and Technology, Xidian University , Xi'an, Shaanxi 710071, China.|Key Laboratory of Molecular Imaging of Chinese Academy of Sciences, Institute of Automation, Chinese Academy of Sciences , Beijing 100190, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Tian
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D050091:Dendrimers / Q000008:administration & dosage; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D018014:Gene Transfer Techniques; D006108:Graphite / Q000008:administration & dosage* / Q000737:chemistry; D006801:Humans; D008055:Lipids / Q000627:therapeutic use; D051379:Mice; D035683:MicroRNAs / Q000037:antagonists & inhibitors / Q000235:genetics*; D057054:Molecular Imaging; D053758:Nanoparticles / Q000008:administration & dosage* / Q000737:chemistry; D009369:Neoplasms / Q000235:genetics / Q000628:therapy*; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage* / Q000235:genetics / Q000627:therapeutic use; D051059:PTEN Phosphohydrolase / Q000096:biosynthesis; D011092:Polyethylene Glycols / Q000008:administration & dosage; D023041:Xenograft Model Antitumor Assays
chemical_list: D050091:Dendrimers; D008055:Lipids; C086724:Lipofectamine; C524940:MIRN21 microRNA, human; D035683:MicroRNAs; D016376:Oligonucleotides, Antisense; C104700:PAMAM Starburst; D011092:Polyethylene Glycols; D006108:Graphite; D051059:PTEN Phosphohydrolase; C494929:PTEN protein, human
keywords: bioluminescence imaging; dendrimer; graphene oxide; luciferase reporter; miRNA delivery