March 2012
Volume 53, Issue 14
Free
ARVO Annual Meeting Abstract  |   March 2012
Effect of Morpholino Targeting Flt-1 in Mice Corneal Keratoplasty Model
Author Affiliations & Notes
  • Yang Kyung Cho
    Ophthalmology, St.Vincent's hospital, The Catholic University of Korea, Suwon, Republic of Korea
    Moran Eye Center, University of Utah, Salt Lake City, Utah
  • Hironori Uehara
    Moran Eye Center, University of Utah, Salt Lake City, Utah
  • Jason R. Young
    Moran Eye Center, University of Utah, Salt Lake City, Utah
  • Wei Huang
    Moran Eye Center, University of Utah, Salt Lake City, Utah
  • Ling Luo
    Moran Eye Center, University of Utah, Salt Lake City, Utah
    Ophthalmology, The 306 Hospital, Beijing, China
  • Xiaohui Zhang
    Moran Eye Center, University of Utah, Salt Lake City, Utah
  • Bonnie Archer
    Moran Eye Center, University of Utah, Salt Lake City, Utah
  • Jacquelyn Simonis
    Moran Eye Center, University of Utah, Salt Lake City, Utah
  • Thomas Olson
    Moran Eye Center, University of Utah, Salt Lake City, Utah
  • Bala Ambati
    Moran Eye Center, University of Utah, Salt Lake City, Utah
  • Footnotes
    Commercial Relationships  Yang Kyung Cho, None; Hironori Uehara, None; Jason R. Young, None; Wei Huang, None; Ling Luo, None; Xiaohui Zhang, None; Bonnie Archer, None; Jacquelyn Simonis, None; Thomas Olson, None; Bala Ambati, None
  • Footnotes
    Support  NIHEY017950
Investigative Ophthalmology & Visual Science March 2012, Vol.53, 2950. doi:
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      Yang Kyung Cho, Hironori Uehara, Jason R. Young, Wei Huang, Ling Luo, Xiaohui Zhang, Bonnie Archer, Jacquelyn Simonis, Thomas Olson, Bala Ambati; Effect of Morpholino Targeting Flt-1 in Mice Corneal Keratoplasty Model. Invest. Ophthalmol. Vis. Sci. 2012;53(14):2950.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : Morpholinos are synthetic molecules similar to DNA oligonucleotides which can bind mRNA and pre-mRNA and sterically block the molecular machinery of translation or alternative splicing. We sought to determine whether morpholinos can target the alternative splicing of Flt-1 pre-mRNA to upregulate the production of sFlt, limiting neovascularization and lymphangiogenesis resulting in increased graft survival in a murine corneal penetrating keratoplasty(PK) model.

Methods: : We used a murine corneal keratoplasty model to investigate the effect of a morpholino that promotes sFlt to suppress neovascularization and lymphangiogenesis to increase graft survival. We performed mice corneal penetrating keratoplasty in one eye of each Balb/C mouse (C57BL6 mice as donors) and injected 40ng/ul of Flt morpholino(n=16), nonspecific (STD) morpholino(n=18), and PBS(n=11) into the subconjucntival space on the day of PK and weekly until postoperative 4 weeks. Corneal opacity was graded to evaluate graft survival weekly through postoperative week 8, at which point the corneas were harvested. After flat mounting the harvested corneas, we performed immunohistochemistry and fluorescent microscopy to digitally quantify neovascularization. We compared the area of neovascularization and lymphangiogenesis to the total corneal area and graft area

Results: : Flt morpholino increased graft survival compared to the PBS group (p=0.043).Flt morpholino group showed less neovascularized area in both total cornea (0.2267±0.0139) and graft (0.1005±0.0173) cornea when compared with STD the morpholino group (0.2771±0.0257 in total cornea, 0.2190±0.0328 in graft cornea, respectively) and PBS group (0.2777±0.0220 in total cornea, 0.1883±0.0384 in graft cornea, respectively) (all p’s<0.05). The Flt morpholino group showed less lymphangiogensis in both total cornea (0.0780±0.0095) and graft cornea (0.0719±0.0109) compared with the STD morpholino group (0.1005±0.0114 in total cornea, 0.1258±0.0198 in graft cornea) and PBS group (0.1199±0.0078 in total cornea, 0.1231±0.0165 in graft cornea) (all p’s<0.05).

Conclusions: : We found that morpholinos promoting sFlt reduced angiogenesis and lymphangiogenesis compared with STD morpholino and PBS in a murine corneal penetrating keratoplasty model and increased graft survival compared with PBS.

Keywords: cornea: basic science • neovascularization • vascular endothelial growth factor 
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