July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
Silk-Derived Protein-4 (SDP-4) Inhibits Nuclear Factor Kappa B (NF-κB) Inflammatory Signaling that Underlies Dry Eye Disease (DED)
Author Affiliations & Notes
  • David W Infanger
    Research and Development, Silk Technologies, Ltd. , Plymouth, Minnesota, United States
  • Waleed Abdel-Naby
    Research and Development, Silk Technologies, Ltd. , Plymouth, Minnesota, United States
  • Jinniece J Kalal
    Research and Development, Silk Technologies, Ltd. , Plymouth, Minnesota, United States
  • Nicholas B Paulson
    Research and Development, Silk Technologies, Ltd. , Plymouth, Minnesota, United States
  • Yue Bai
    Research and Development, Silk Technologies, Ltd. , Plymouth, Minnesota, United States
  • Brian D Lawrence
    Research and Development, Silk Technologies, Ltd. , Plymouth, Minnesota, United States
  • Footnotes
    Commercial Relationships   David Infanger, Silk Technologies, Ltd. (E); Waleed Abdel-Naby, Silk Technologies, Ltd. (F); Jinniece Kalal, Silk Technologies, Ltd. (E); Nicholas Paulson, Silk Technologies, Ltd. (E); Yue Bai, Silk Technologies, Ltd. (E); Brian Lawrence, Silk Technologies, Ltd. (E)
  • Footnotes
    Support  Army SBIR P1 W81XWH-17-C-0150
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 2820. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      David W Infanger, Waleed Abdel-Naby, Jinniece J Kalal, Nicholas B Paulson, Yue Bai, Brian D Lawrence; Silk-Derived Protein-4 (SDP-4) Inhibits Nuclear Factor Kappa B (NF-κB) Inflammatory Signaling that Underlies Dry Eye Disease (DED). Invest. Ophthalmol. Vis. Sci. 2019;60(9):2820.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : Chronic inflammation governed by the NF-κB signaling pathway is a hallmark of DED that contributes to the development of patient symptomology. Previous work has demonstrated that solubilized proteins derived from the Bombyx mori silkworm cocoon can be utilized to reduce DED related biomarkers in animal models. The data presented here support the hypothesis that the silk-derived biotherapeutic candidate, SDP-4, effectively inhibits NF-κB activation and downstream inflammatory signaling in vitro in both human corneal limbal-epithelial (HCLE) and embryonic kidney (HEK293) cell lines.

Methods : SDP-4 was prepared by performing an alkaline extraction on Bombyx mori silkworm cocoons, which was followed by dissolution of extracted fibers in concentrated LiBr solution under heat and pressure. Final purification of the 20kDa SDP-4 molecule was achieved using membrane dialysis. NF-κB-mediated gene expression was stimulated in HCLEs using TNF-α (1ng/mL) in the presence/absence of SDP-4 (up to 1%) for up to 6-hours, and expression of inflammatory cytokines measured using quantitative PCR and ELISA. Coculture immune cell (HL-60) migration assays were performed to evaluate the functional impact of impaired HCLE signaling. To determine whether SDP-4 inhibits transcription factor activation directly, a HEK reporter cell line was used to quantitate NF-κB-driven chemiluminescence in the presence or absence of SDP-4.

Results : SDP-4 significantly reduced (p<0.05, n=3) TNF-α induced expression of the inflammatory genes TNF-α, MMP-9, IL-1a/b and IL-8. Furthermore, there was a dose-dependent inhibition of IL-8 production by over 50% with increasing SDP-4 concentration. This reduction in cytokine release by HCLEs was functionally sufficient to significantly inhibit (p<0.05, n=3) activation/migration of immune-like HL-60 cells. Lastly, SDP-4 demonstrated a dose-dependent reduction in NF-κB-mediated chemiluminescence in stimulated reporter cells.

Conclusions : These studies support that SDP-4 inhibits in vitro corneal inflammatory signaling mediated by NF-κB. Additionally, this reduced expression was effective to inhibit activation of immune-like cells, which may provide a novel opportunity to inhibit chronic inflammatory signaling in DED.

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

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×