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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)
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.
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.
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.
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.
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