June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Assessing Inhibition of HUVEC Migration by a Novel VEGF-A and Angiopoietin-2 Bispecific Protein (RO-634)
Author Affiliations & Notes
  • John Kunzeman
    Southern Illinois University School of Medicine, Springfield, Illinois, United States
  • Jeffrey L Olson
    University of Colorado - Anschutz Medical Campus, Aurora, Colorado, United States
  • Anthony Jones
    University of Colorado - Anschutz Medical Campus, Aurora, Colorado, United States
  • Josh Morgenstern
    University of Colorado - Anschutz Medical Campus, Aurora, Colorado, United States
  • Anne Strong
    University of Colorado - Anschutz Medical Campus, Aurora, Colorado, United States
  • Steven Droho
    University of Colorado - Anschutz Medical Campus, Aurora, Colorado, United States
  • Shaun Bevers
    Department of Structural Biology and Biochemistry, University of Colorado Denver School of Medicine, Aurora, Colorado, United States
  • Niklaus Mueller
    University of Colorado - Anschutz Medical Campus, Aurora, Colorado, United States
  • Michael Huvard
    University of Colorado - Anschutz Medical Campus, Aurora, Colorado, United States
  • Li Xu
    Independent Research Consultant, California, United States
  • Peter K Kaiser
    Cleveland Clinic Cole Eye Institute, Cleveland, Ohio, United States
  • Arshad M. Khanani
    Sierra Eye Associates, Reno, Nevada, United States
    University of Nevada Reno School of Medicine, Reno, Nevada, United States
  • Jeffrey S Heier
    OCB, Boston, Massachusetts, United States
  • Nikhil Gupta
    Glenwood High School, Chatham, Illinois, United States
  • Alina Sinha
    University of Missouri Kansas City School of Medicine, Kansas City, Missouri, United States
  • Ramanath Bhandari
    Eye Institute, Springfield Clinic LLP, Springfield, Illinois, United States
  • Footnotes
    Commercial Relationships   John Kunzeman None; Jeffrey Olson RevOpsis Therapeutics, 2C Tech, Code O (Owner); Anthony Jones None; Josh Morgenstern None; Anne Strong None; Steven Droho None; Shaun Bevers None; Niklaus Mueller None; Michael Huvard None; Li Xu RevOpsis Therapeutics, Protagonist Therapeutics, Code C (Consultant/Contractor); Peter Kaiser AffaMed, Allergan, Bayer, Regeneron, Novartis, Kanghong, RevOpsis, Boerenger Ingelhein, Kodiak, RegenxBio, Code C (Consultant/Contractor), RevOpsis, Code I (Personal Financial Interest); Arshad Khanani 4DMT, Adverum, Allergan, Genentech, Regeneron, Novartis, Kanghong, RevOpsis, Kodiak, RegenxBio, Code C (Consultant/Contractor), RevOpsis, Code I (Personal Financial Interest); Jeffrey Heier 2020 Onsite, 4DMT, Abpro, Adverum, Allegro, Allergan, Annexon, Apellis, Asclepix, Aviceda, BVT, DTx, Gemini, Genetech/Roche, Graybug, Gyroscope, iRenix, Iveric, Johnson & Johnson, Kang Horn, NGM, Notal Vision, Novartis, Ocular Therapeutix, Ocuphire, OcuTerra, Oriole, Oxurion, Regeneron, Regenxbio, Relay Therapeutics, RetinAI, Retrotope, Roche, Stealth Biotherapeutics, Surrozen, Thea, Unity Bio, Verseon, Code C (Consultant/Contractor), Aldeyra, Apellis, Asclepix, Bayer, Genentech, Gyroscope, Iveric, Janssen R&D, Kanghong, Kodiak, NGM, Notal Vision, Novartis, Regeneron, Regenxbio, Stealth, Code F (Financial Support), Adverum, Aldeyra, Allegro, Aviceda, DTx Pharma, jCyte, Ocular Therapeutix, Vinci, Vitranu, Code I (Personal Financial Interest), Ocular Therapeutix, Code S (non-remunerative); Nikhil Gupta None; Alina Sinha None; Ramanath Bhandari Regeneron, Kodiak Biosciences, Code C (Consultant/Contractor), RevOpsis Therapeutics, Code O (Owner)
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 1325 – F0159. doi:
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      John Kunzeman, Jeffrey L Olson, Anthony Jones, Josh Morgenstern, Anne Strong, Steven Droho, Shaun Bevers, Niklaus Mueller, Michael Huvard, Li Xu, Peter K Kaiser, Arshad M. Khanani, Jeffrey S Heier, Nikhil Gupta, Alina Sinha, Ramanath Bhandari; Assessing Inhibition of HUVEC Migration by a Novel VEGF-A and Angiopoietin-2 Bispecific Protein (RO-634). Invest. Ophthalmol. Vis. Sci. 2022;63(7):1325 – F0159.

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

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Abstract

Purpose : To investigate how a novel vascular endothelial growth-factor A (VEGF-A) and angiopoietin-2 (Ang-2) bispecific protein (surrobody), RO-634, inhibits human umbilical vein endothelial cell (HUVECs) migration by assessing wound closure post exposure to VEGF-A in laboratory models.

Methods : HUVEC cells were purchased and grown in a 5% CO2 humidified incubator at 37°C. Endothelial Cell Medium was used for optimal proliferation. Culture-inserts were used to evaluate HUVEC migration potential. Thirty-five thousand HUVEC cells were seeded into a bifurcated chamber and incubated overnight. Eighteen hours after initial seeding, the culture inserts were removed and cells were washed 3x with 1x PBS. After washing, media containing 40 ng of VEGF-A was added along with either bevacizumab, aflibercept, or RO-634. Cells were incubated with the desired medium for 12 hours. The percent change in wound closure was calculated using an image of the same area over the two designated timepoints. This process was repeated in triplicate.

Results : At 12 hours, HUVEC cells exposed to 40 ng of VEGF-A and aflibercept demonstrated a 70% reduction in VEGF-induced HUVEC migration compared to no treatment. HUVEC cells exposed to 40 ng of VEGF-A and bevacizumab demonstrated a 22% reduction in VEGF-induced HUVEC migration compared to no treatment. HUVEC cells exposed to 40 ng of VEGF-A and RO-634 demonstrated a 68% reduction in VEGF-induced HUVEC migration compared to no treatment. The difference in reduction in VEGF-induced HUVEC migration between aflibercept and RO-634 was insignificant (p=0.93). The results are shown in figure 1.

Conclusions : This data suggests that novel bispecific protein (surrobody), RO-634, is effective at preventing HUVEC migration post exposure to VEGF-A and is superior at preventing cell migration when compared with bevacizumab in this experiment. There is no statistically significant difference between aflibercept and RO-634 in preventing cell migration of endothelial cells.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.

 

Figure 1: Comparison of wound closure in HUVEC cells after exposure to 40 ng of VEGF-A and either Bevacizumab, Aflibercept, or RO-634 at 0 and 12 hours.

Figure 1: Comparison of wound closure in HUVEC cells after exposure to 40 ng of VEGF-A and either Bevacizumab, Aflibercept, or RO-634 at 0 and 12 hours.

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