Nanoparticle-mediated miR200-b delivery for the treatment of diabetic retinopathy.
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.
Department of Ophthalmology, University of North Carolina, Chapel Hill, NC 27599, USA.
Rajendra Narayan
RN
Mitra
Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA.
Chance A
CA
Nichols
Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA.
Junjing
J
Guo
Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA.
Rasha
R
Makkia
Copernicus Therapeutics, Incorporated, Cleveland, OH 44106, USA.
Mark J
MJ
Cooper
Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA; Department of Biomedical Engineering, University of Houston, Houston, TX 77204, USA.
Muna I
MI
Naash
Department of Ophthalmology, University of North Carolina, Chapel Hill, NC 27599, USA; Carolina Institute for NanoMedicine, University of North Carolina, Chapel Hill, NC 27599, USA; Division of Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, NC 27599, USA. Electronic address: zongchao@med.unc.edu.
Zongchao
Z
Han
D000818:Animals; D002460:Cell Line; D002465:Cell Movement; D003921:Diabetes Mellitus, Experimental / Q000150:complications; D003930:Diabetic Retinopathy / Q000188:drug therapy* / Q000209:etiology; D005260:Female; D015870:Gene Expression; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D058449:Intravitreal Injections; D008297:Male; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D035683:MicroRNAs / Q000008:administration & dosage* / Q000235:genetics; D053758:Nanoparticles / Q000737:chemistry*; D018919:Neovascularization, Physiologic; D010455:Peptides / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D014162:Transfection / Q000379:methods; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000378:metabolism; D014945:Wound Healing
D035683:MicroRNAs; D010455:Peptides; D011092:Polyethylene Glycols; D040301:Vascular Endothelial Growth Factor Receptor-2
Angiogenesis; Diabetic retinopathy; Nanoparticles; and VEGF receptors; miR200-b