Sustained delivery of proangiogenic microRNA-132 by nanoparticle transfection improves endothelial cell transplantation.
Transplantation of endothelial cells (ECs) for therapeutic vascularization or tissue engineering is a promising method for increasing tissue perfusion. Here, we report on a new approach for enhanced EC transplantation using targeted nanoparticle transfection to deliver proangiogenic microRNA-132 (miR-132) to cultured ECs before their transplantation, thereby sensitizing cells to the effects of endogenous growth factors. We synthesized biodegradable PLGA polymer nanoparticles (NPs) that were loaded with miR-132 and coated with cyclic RGD (cRGD) peptides that target integrin αvβ3 expressed on cultured human umbilical vein ECs (HUVECs), increasing NP uptake through clathrin-coated pits. Unlike previously reported NPs for miR delivery, these NPs slowly release RNA for several weeks. The endocytosed NPs remain in clathrin-coated vesicles from which they mediate intracellular delivery of siRNA or miRNA. Transfection of HUVECs with miR-132 enhances growth factor-induced proliferation and migration in 2D culture, producing a 1.8- and 5-fold increase, respectively. However, while the effects of conventional transfection were short-lived, NP transfection produced protein knockdown and biological effects that were significantly longer in duration (≥ 6 d). Transfection of HUVECs with miR-132 NP resulted in a 2-fold increase in the number of microvessels per square millimeter compared to lipid after transplantation into immunodeficient mice and led to a higher number of mural cell-invested vessels than control transfection. These data suggest that sustained delivery of miR-132 encapsulated in a targeted biodegradable polymer NP is a safe and efficient strategy to improve EC transplantation and vascularization.
2Department of Immunobiology, PO Box 208089, New Haven, CT 06520-8089, USA. jordan.pober@yale.edu.
Julie
J
Devalliere
William G
WG
Chang
Jillian W
JW
Andrejecsk
Parwiz
P
Abrahimi
Christopher J
CJ
Cheng
Dan
D
Jane-wit
W Mark
WM
Saltzman
Jordan S
JS
Pober
D000818:Animals; D015153:Blotting, Western; D005260:Female; D005434:Flow Cytometry; D061307:Human Umbilical Vein Endothelial Cells / Q000637:transplantation; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D035683:MicroRNAs / Q000008:administration & dosage* / Q000235:genetics*; D018613:Microscopy, Confocal; D053758:Nanoparticles / Q000008:administration & dosage*; D036103:Nanotechnology / Q000379:methods; D018919:Neovascularization, Physiologic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D023822:Tissue Engineering; D014162:Transfection / Q000379:methods
D035683:MicroRNAs
controlled release; nanotechnology; neovascularization; tissue engineering