Utilizing targeted gene therapy with nanoparticles binding alpha v Beta 3 for imaging and treating choroidal Neovascularization

Hani Salehi-Had, Mi In Roh, Andrea Giani, Toshio Hisatomi, Shintaro Nakao, Ivana K. Kim, Evangelos S. Gragoudas, Demetrios Vavvas, Samira Guccione, Joan W. Miller

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Purpose: The integrin αvβ3 is differentially expressed on neovascular endothelial cells. We investigated whether a novel intravenously injectable αvβ3 integrin-ligand coupled nanoparticle (NP) can target choroidal neovascular membranes (CNV) for imaging and targeted gene therapy. Methods: CNV lesions were induced in rats using laser photocoagulation. The utility of NP for in vivo imaging and gene delivery was evaluated by coupling the NP with a green fluorescing protein plasmid (NP-GFPg). Rhodamine labeling (Rd-NP) was used to localize NP in choroidal flatmounts. Rd-NP-GFPg particles were injected intravenously on weeks 1, 2, or 3. In the treatment arm, rats received NP containing a dominant negative Raf mutant gene (NP-ATPμ-Raf) on days 1, 3, and 5. The change in CNV size and leakage, and TUNEL positive cells were quantified. Results: GFP plasmid expression was seen in vivo up to 3 days after injection of Rd-NP-GFPg. Choroidal flatmounts confirmed the localization of the NP and the expression of GFP plasmid in the CNV. Treating the CNV with NP-ATPμ-Raf decreased the CNV size by 42% (P<0.001). OCT analysis revealed that the reduction of CNV size started on day 5 and reached statistical significance by day 7. Fluorescein angiography grading showed significantly less leakage in the treated CNV (P<0.001). There were significantly more apoptotic (TUNEL-positive) nuclei in the treated CNV. Conclusion: Systemic administration of αvβ3 targeted NP can be used to label the abnormal blood vessels of CNV for imaging. Targeted gene delivery with NP-ATPμ-Raf leads to a reduction in size and leakage of the CNV by induction of apoptosis in the CNV.

Original languageEnglish
Article numbere18864
JournalPloS one
Volume6
Issue number4
DOIs
Publication statusPublished - May 12 2011

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Gene therapy
Choroidal Neovascularization
gene therapy
nanoparticles
angiogenesis
Genetic Therapy
Nanoparticles
image analysis
Imaging techniques
Membranes
plasmids
Plasmids
Genes
Adenosine Triphosphate
integrins
In Situ Nick-End Labeling
gene transfer
Integrins
Rats
Injections

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

Cite this

Utilizing targeted gene therapy with nanoparticles binding alpha v Beta 3 for imaging and treating choroidal Neovascularization. / Salehi-Had, Hani; Roh, Mi In; Giani, Andrea; Hisatomi, Toshio; Nakao, Shintaro; Kim, Ivana K.; Gragoudas, Evangelos S.; Vavvas, Demetrios; Guccione, Samira; Miller, Joan W.

In: PloS one, Vol. 6, No. 4, e18864, 12.05.2011.

Research output: Contribution to journalArticle

Salehi-Had, H, Roh, MI, Giani, A, Hisatomi, T, Nakao, S, Kim, IK, Gragoudas, ES, Vavvas, D, Guccione, S & Miller, JW 2011, 'Utilizing targeted gene therapy with nanoparticles binding alpha v Beta 3 for imaging and treating choroidal Neovascularization', PloS one, vol. 6, no. 4, e18864. https://doi.org/10.1371/journal.pone.0018864
Salehi-Had, Hani ; Roh, Mi In ; Giani, Andrea ; Hisatomi, Toshio ; Nakao, Shintaro ; Kim, Ivana K. ; Gragoudas, Evangelos S. ; Vavvas, Demetrios ; Guccione, Samira ; Miller, Joan W. / Utilizing targeted gene therapy with nanoparticles binding alpha v Beta 3 for imaging and treating choroidal Neovascularization. In: PloS one. 2011 ; Vol. 6, No. 4.
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abstract = "Purpose: The integrin αvβ3 is differentially expressed on neovascular endothelial cells. We investigated whether a novel intravenously injectable αvβ3 integrin-ligand coupled nanoparticle (NP) can target choroidal neovascular membranes (CNV) for imaging and targeted gene therapy. Methods: CNV lesions were induced in rats using laser photocoagulation. The utility of NP for in vivo imaging and gene delivery was evaluated by coupling the NP with a green fluorescing protein plasmid (NP-GFPg). Rhodamine labeling (Rd-NP) was used to localize NP in choroidal flatmounts. Rd-NP-GFPg particles were injected intravenously on weeks 1, 2, or 3. In the treatment arm, rats received NP containing a dominant negative Raf mutant gene (NP-ATPμ-Raf) on days 1, 3, and 5. The change in CNV size and leakage, and TUNEL positive cells were quantified. Results: GFP plasmid expression was seen in vivo up to 3 days after injection of Rd-NP-GFPg. Choroidal flatmounts confirmed the localization of the NP and the expression of GFP plasmid in the CNV. Treating the CNV with NP-ATPμ-Raf decreased the CNV size by 42{\%} (P<0.001). OCT analysis revealed that the reduction of CNV size started on day 5 and reached statistical significance by day 7. Fluorescein angiography grading showed significantly less leakage in the treated CNV (P<0.001). There were significantly more apoptotic (TUNEL-positive) nuclei in the treated CNV. Conclusion: Systemic administration of αvβ3 targeted NP can be used to label the abnormal blood vessels of CNV for imaging. Targeted gene delivery with NP-ATPμ-Raf leads to a reduction in size and leakage of the CNV by induction of apoptosis in the CNV.",
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AU - Giani, Andrea

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AU - Nakao, Shintaro

AU - Kim, Ivana K.

AU - Gragoudas, Evangelos S.

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