title: Nanoparticle-mediated expression of a Wnt pathway inhibitor ameliorates ocular neovascularization.
abstract: OBJECTIVE
  The deficiency of very low-density lipoprotein receptor resulted in Wnt signaling activation and neovascularization in the retina. The present study sought to determine whether the very low-density lipoprotein receptor extracellular domain (VLN) is responsible for the inhibition of Wnt signaling in ocular tissues.
  
  
  RESULTS
  A plasmid expressing the soluble VLN was encapsulated with poly(lactide-co-glycolide acid) to form VLN nanoparticles (VLN-NP). Nanoparticles containing a plasmid expressing the low-density lipoprotein receptor extracellular domain nanoparticle were used as negative control. MTT, modified Boyden chamber, and Matrigel (™) assays were used to evaluate the inhibitory effect of VLN-NP on Wnt3a-stimulated endothelial cell proliferation, migration, and tube formation. Vldlr(-/-) mice, oxygen-induced retinopathy, and alkali burn-induced corneal neovascularization models were used to evaluate the effect of VLN-NP on ocular neovascularization. Wnt reporter mice (BAT-gal), Western blotting, and luciferase assay were used to evaluate Wnt pathway activity. Our results showed that VLN-NP specifically inhibited Wnt3a-induced endothelial cell proliferation, migration, and tube formation. Intravitreal injection of VLN-NP inhibited abnormal neovascularization in Vldlr(-/-), oxygen-induced retinopathy, and alkali burn-induced corneal neovascularization models, compared with low-density lipoprotein receptor extracellular domain nanoparticle. VLN-NP significantly inhibited the phosphorylation of low-density lipoprotein receptor-related protein 6, the accumulation of β-catenin, and the expression of vascular endothelial growth factor in vivo and in vitro.
  
  
  CONCLUSIONS
  Taken together, these results suggest that the soluble VLN is a negative regulator of the Wnt pathway and has antiangiogenic activities. Nanoparticle-mediated expression of VLN may thus represent a novel therapeutic approach to treat pathological ocular angiogenesis and potentially other vascular diseases affected by Wnt signaling.
authors: From the Department of Ophthalmology, Shanghai First People's Hospital Affiliated to Shanghai Jiaotong University, Shanghai, China (Z.W., X.X.); Department of Physiology, University of Oklahoma Health Sciences Center, Oklahoma City (Z.W., R.C., K.L., L.D., J.-x.M.); Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Denver, Aurora (P.T., U.B.K.); Department of Ophthalmology, Xiangya Hospital, Central South University, Changsha, China (L.D.); and Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (J.C.).
authors: Zhongxiao
authors: Z
authors: Wang
authors: From the Department of Ophthalmology, Shanghai First People's Hospital Affiliated to Shanghai Jiaotong University, Shanghai, China (Z.W., X.X.); Department of Physiology, University of Oklahoma Health Sciences Center, Oklahoma City (Z.W., R.C., K.L., L.D., J.-x.M.); Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Denver, Aurora (P.T., U.B.K.); Department of Ophthalmology, Xiangya Hospital, Central South University, Changsha, China (L.D.); and Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (J.C.).
authors: Rui
authors: R
authors: Cheng
authors: From the Department of Ophthalmology, Shanghai First People's Hospital Affiliated to Shanghai Jiaotong University, Shanghai, China (Z.W., X.X.); Department of Physiology, University of Oklahoma Health Sciences Center, Oklahoma City (Z.W., R.C., K.L., L.D., J.-x.M.); Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Denver, Aurora (P.T., U.B.K.); Department of Ophthalmology, Xiangya Hospital, Central South University, Changsha, China (L.D.); and Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (J.C.).
authors: Kyungwon
authors: K
authors: Lee
authors: From the Department of Ophthalmology, Shanghai First People's Hospital Affiliated to Shanghai Jiaotong University, Shanghai, China (Z.W., X.X.); Department of Physiology, University of Oklahoma Health Sciences Center, Oklahoma City (Z.W., R.C., K.L., L.D., J.-x.M.); Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Denver, Aurora (P.T., U.B.K.); Department of Ophthalmology, Xiangya Hospital, Central South University, Changsha, China (L.D.); and Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (J.C.).
authors: Puneet
authors: P
authors: Tyagi
authors: From the Department of Ophthalmology, Shanghai First People's Hospital Affiliated to Shanghai Jiaotong University, Shanghai, China (Z.W., X.X.); Department of Physiology, University of Oklahoma Health Sciences Center, Oklahoma City (Z.W., R.C., K.L., L.D., J.-x.M.); Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Denver, Aurora (P.T., U.B.K.); Department of Ophthalmology, Xiangya Hospital, Central South University, Changsha, China (L.D.); and Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (J.C.).
authors: Lexi
authors: L
authors: Ding
authors: From the Department of Ophthalmology, Shanghai First People's Hospital Affiliated to Shanghai Jiaotong University, Shanghai, China (Z.W., X.X.); Department of Physiology, University of Oklahoma Health Sciences Center, Oklahoma City (Z.W., R.C., K.L., L.D., J.-x.M.); Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Denver, Aurora (P.T., U.B.K.); Department of Ophthalmology, Xiangya Hospital, Central South University, Changsha, China (L.D.); and Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (J.C.).
authors: Uday B
authors: UB
authors: Kompella
authors: From the Department of Ophthalmology, Shanghai First People's Hospital Affiliated to Shanghai Jiaotong University, Shanghai, China (Z.W., X.X.); Department of Physiology, University of Oklahoma Health Sciences Center, Oklahoma City (Z.W., R.C., K.L., L.D., J.-x.M.); Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Denver, Aurora (P.T., U.B.K.); Department of Ophthalmology, Xiangya Hospital, Central South University, Changsha, China (L.D.); and Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (J.C.).
authors: Jing
authors: J
authors: Chen
authors: From the Department of Ophthalmology, Shanghai First People's Hospital Affiliated to Shanghai Jiaotong University, Shanghai, China (Z.W., X.X.); Department of Physiology, University of Oklahoma Health Sciences Center, Oklahoma City (Z.W., R.C., K.L., L.D., J.-x.M.); Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Denver, Aurora (P.T., U.B.K.); Department of Ophthalmology, Xiangya Hospital, Central South University, Changsha, China (L.D.); and Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (J.C.). jian-xing-ma@ouhsc.edu drxuxun@sjtu.edu.cn.
authors: Xun
authors: X
authors: Xu
authors: From the Department of Ophthalmology, Shanghai First People's Hospital Affiliated to Shanghai Jiaotong University, Shanghai, China (Z.W., X.X.); Department of Physiology, University of Oklahoma Health Sciences Center, Oklahoma City (Z.W., R.C., K.L., L.D., J.-x.M.); Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Denver, Aurora (P.T., U.B.K.); Department of Ophthalmology, Xiangya Hospital, Central South University, Changsha, China (L.D.); and Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, MA (J.C.). jian-xing-ma@ouhsc.edu drxuxun@sjtu.edu.cn.
authors: Jian-Xing
authors: JX
authors: Ma
mesh_terms: D000818:Animals; D002465:Cell Movement; D049109:Cell Proliferation; D002478:Cells, Cultured; D003315:Cornea / Q000098:blood supply*; D016510:Corneal Neovascularization / Q000235:genetics / Q000378:metabolism / Q000503:physiopathology / Q000517:prevention & control*; D004195:Disease Models, Animal; D006801:Humans; D058449:Intravitreal Injections; D019344:Lactic Acid / Q000737:chemistry*; D060488:Low Density Lipoprotein Receptor-Related Protein-6 / Q000378:metabolism; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D053758:Nanoparticles; D010766:Phosphorylation; D011100:Polyglycolic Acid / Q000737:chemistry*; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D034622:RNA Interference; D017207:Rats, Sprague-Dawley; D011973:Receptors, LDL / Q000235:genetics / Q000378:metabolism*; D015861:Retinal Neovascularization / Q000235:genetics / Q000378:metabolism / Q000503:physiopathology / Q000517:prevention & control*; D012171:Retinal Vessels / Q000378:metabolism* / Q000503:physiopathology; D013997:Time Factors; D014162:Transfection / Q000379:methods*; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism; D060449:Wnt Signaling Pathway; D060509:Wnt3A Protein / Q000235:genetics / Q000378:metabolism*; D051176:beta Catenin / Q000378:metabolism
chemical_list: D060488:Low Density Lipoprotein Receptor-Related Protein-6; D011973:Receptors, LDL; C036053:VLDL receptor; D042461:Vascular Endothelial Growth Factor A; D060509:Wnt3A Protein; D051176:beta Catenin; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid
keywords: VLDLR; Wnt; eye; nanoparticle; neovascularization
references: Diabetes. 2012 Nov;61(11):2948-57
references: 22891217
references: J Clin Invest. 2009 Mar;119(3):611-23
references: 19188685
references: PLoS One. 2011;6(1):e16712
references: 21304961
references: J Burn Care Res. 2014 May-Jun;35(3):261-8
references: 23877138
references: Ann Med. 2006;38(7):450-71
references: 17101537
references: Nat Struct Mol Biol. 2011 Nov;18(11):1204-10
references: 21984209
references: Circulation. 2011 Oct 25;124(17):1871-81
references: 21969016
references: Front Biosci. 2006;11:2442-64
references: 16720326
references: Diabetes Care. 2004 Oct;27(10):2568-9; author reply 2569
references: 15451946
references: Exp Cell Res. 2013 May 15;319(9):1317-23
references: 23291327
references: J Clin Invest. 1993 Aug;92(2):883-93
references: 8349823
references: Ann Med. 2012 Feb;44(1):1-17
references: 21284527
references: Diabetes. 2009 Aug;58(8):1902-13
references: 19491211
references: Diabetes. 2013 Dec;62(12):4228-38
references: 23884893
references: FASEB J. 2002 Aug;16(10):1217-26
references: 12153989
references: Invest Ophthalmol Vis Sci. 2009 Feb;50(2):844-50
references: 18936153
references: Nat Commun. 2014;5:3930
references: 24852848
references: J Mol Med (Berl). 2013 Mar;91(3):311-21
references: 23329331
references: Circ Res. 2010 Oct 15;107(8):943-52
references: 20947863
references: Invest Ophthalmol Vis Sci. 2013 Jan;54(1):141-54
references: 23211829
references: Endocrinology. 1995 Jan;136(1):340-8
references: 7828550
references: FEBS Lett. 1999 May 14;451(1):56-62
references: 10356983
references: Nat Rev Cancer. 2008 May;8(5):387-98
references: 18432252
references: Asian Pac J Cancer Prev. 2014;15(2):517-35
references: 24568455
references: Nature. 2001 May 17;411(6835):321-5
references: 11357136
references: Br J Ophthalmol. 2005 Dec;89(12):1627-30
references: 16299144
references: Retina. 2003 Aug;23(4):518-22
references: 12972764
references: J Biol Chem. 2007 Nov 23;282(47):34420-8
references: 17890782
references: Genomics. 1994 Mar 15;20(2):298-300
references: 8020981
references: Arch Ophthalmol. 2007 Jun;125(6):795-803
references: 17562991
references: Int J Ophthalmol. 2010;3(4):304-7
references: 22553579
references: Int J Nanomedicine. 2013;8:495-503
references: 23439842
references: Am J Pathol. 2012 Aug;181(2):376-9
references: 22749677
references: Macromol Biosci. 2012 May;12(5):608-20
references: 22508445
references: J Cell Sci. 2014 Nov 15;127(Pt 22):4857-69
references: 25271056
references: EMBO Rep. 2013 May;14(5):422-33
references: 23598517
references: PLoS One. 2011;6(2):e16733
references: 21364932
references: Annu Rev Biochem. 1994;63:601-37
references: 7979249
references: Proc Natl Acad Sci U S A. 2006 Apr 18;103(16):6315-20
references: 16606824