Nanoparticle-mediated miR200-b delivery for the treatment of diabetic retinopathy
| Autor: | Rajendra Narayan Mitra, Muna I. Naash, Junjing Guo, Zongchao Han, Rasha Makkia, Chance A. Nichols, Mark J. Cooper |
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| Rok vydání: | 2016 |
| Předmět: |
Male
0301 basic medicine Angiogenesis Gene Expression Neovascularization Physiologic Pharmaceutical Science Mice Transgenic Biology Transfection Article Cell Line Diabetes Mellitus Experimental Polyethylene Glycols 03 medical and health sciences 0302 clinical medicine Western blot Cell Movement In vivo Gene expression medicine Animals Humans Wound Healing Diabetic Retinopathy medicine.diagnostic_test Gene Transfer Techniques Kinase insert domain receptor Genetic Therapy Vascular Endothelial Growth Factor Receptor-2 Molecular biology Mice Inbred C57BL Endothelial stem cell MicroRNAs 030104 developmental biology Cell culture 030220 oncology & carcinogenesis Intravitreal Injections Nanoparticles Female Peptides |
| Zdroj: | Journal of Controlled Release. 236:31-37 |
| ISSN: | 0168-3659 |
| DOI: | 10.1016/j.jconrel.2016.06.020 |
| Popis: | We recently reported that the Ins2(Akita) mouse is a good model for late-onset diabetic retinopathy. Here, we investigated the effect of miR200-b, a potential anti-angiogenic factor, on VEGF receptor 2 (VEGFR-2) expression and to determine the underlying angiogenic response in mouse endothelial cells, and in retinas from aged Ins2(Akita) mice. MiR200-b and its native flanking sequences were amplified and cloned into a pCAG-eGFP vector directed by the ubiquitous CAG promoter (namely pCAG-miR200-b-IRES-eGFP). The plasmid was compacted by CK30PEG10K into DNA nanoparticles (NPs) for in vivo delivery. Murine endothelial cell line, SVEC4-10, was first transfected with the plasmid. The mRNA levels of VEGF and VEGFR-2 were quantified by qRT-PCR and showed significant reduction in message expression compared with lipofectamine-transfected cells. Transfection of miR200-b suppressed the migration of SVEC4-10 cells. There was a significant inverse correlation between the level of expression of miR200-b and VEGFR-2. Intravitreal injection of miR200-b DNA NPs significantly reduced protein levels of VEGFR-2 as revealed by western blot and markedly suppressed angiogenesis as evaluated by fundus imaging in aged Ins2(Akita) mice even after 3months of post-injection. These findings suggest that NP-mediated miR200-b delivery has negatively regulated VEGFR-2 expression in vivo. |
| Databáze: | OpenAIRE |
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