July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
Role of Insulin-like growth factor binding protein-3 (IGFBP-3) in pathogenesis of herpes stromal keratitis (HSK)
Author Affiliations & Notes
  • Susmit Suvas
    Ophthalmology, Visual and Anatomical Sciences, Wayne State University School of Medicine, Detroit, Michigan, United States
  • Pushpa Rao
    Ophthalmology, Visual and Anatomical Sciences, Wayne State University School of Medicine, Detroit, Michigan, United States
  • Jena J Steinle
    Ophthalmology, Visual and Anatomical Sciences, Wayne State University School of Medicine, Detroit, Michigan, United States
  • Footnotes
    Commercial Relationships   Susmit Suvas, None; Pushpa Rao, None; Jena Steinle, None
  • Footnotes
    Support  NH Grant EY022417 and NH Grant EY029690
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 4627. doi:
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    • Get Citation

      Susmit Suvas, Pushpa Rao, Jena J Steinle; Role of Insulin-like growth factor binding protein-3 (IGFBP-3) in pathogenesis of herpes stromal keratitis (HSK). Invest. Ophthalmol. Vis. Sci. 2019;60(9):4627.

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

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Abstract

Purpose : IGFBP-3 binds to 75% of serum IGF proteins and regulates their bioavailability and IGF-1R mediated signaling. IGFBP-3 also acts in IGF-independent mode, which involves cell surface binding, cytosolic accumulation and nuclear localization of IGFBP-3. IGF-dependent and independent action of IGFBP-3 is known to regulate hemangiogenesis. In this study, we determined the relative contribution of IGFBP-3 in pathogenesis of HSK in a mouse model.

Methods : Uninfected and HSV-1 infected C57BL/6J (B6) mice corneas were homogenized at 5-, 10-, and 15-day post-infection. The corneal lysates were used to perform the angiogenic array blots and quantitation of positive signal on membrane was carried out using Image Studio Lite Ver 4.0 software. RT-qPCR assay, immunofluorescence staining, ELISA assay, Zymography, and flow cytometery was carried out on HSV-1 infected corneal samples. Development of corneal opacity and hemangiogenesis in infected corneas from B6 and IGFBP-3KO mice was measured using hand-held slit lamp microscope.

Results : An elevated level of IGFBP-3 protein was detected in infected corneas at 5-, 10-, and 15-day post-infection than uninfected control corneal samples. Similarly, an elevated mRNA expression of IGFBP-3 was detected in infected corneas at 5-, and 10-day post-infection. Immunofluorescence staining showed an intracellular accumulation of IGFBP-3 protein in corneal epithelial cells. A progressive decrease in IGFBP-3 protein was detected in serum samples obtained from HSV-1 infected mice at different day post-infection. Since IGFBP-3 proteolysis is regulated by matrix metalloproteinases (MMPs) especially MMP-3, we next ascertained the level and enzymatic activity of MMP-3 in infected corneas. An increased level of MMP-3 (by ELISA) and enzymatic activity (Zymography) was detected in corneal samples with HSK. Furthermore, HSV-1 infected corneas of IGFBP-3 KO mice showed a significantly increased scoring of hem-angiogenesis and corneal opacity than control infected B6 mice. This was associated with an increased influx and an enhanced phosphorylation of IGF-1R expressing leukocytes in infected corneas of IGFBP-3 KO mice.

Conclusions : Altogether, our results suggest that IGFBP-3 in HSV-1 infected corneas is likely exerting a protective effect, as their absence causes an increase in opacity and the extent of hemangiogenesis in infected corneas.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

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