The Effect of Genetic and Pharmacologic Loss of Notch4 on Angiogenesis
Session Number
Project ID: BIO 39
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Advisor(s)
Dr. Naiche Adler; University of Illinois at Chicago College of Medicine, Kitajewski Lab
Discipline
Biology
Start Date
22-4-2020 11:30 AM
End Date
22-4-2020 11:55 AM
Abstract
Notch signaling is critical to the formation of blood vessels, and although the role of Notch1 has been well characterized, less is known about Notch4 and the degree to which its function overlaps with Notch1. In order to quantify the effects of removing Notch4, Notch4 mutant mice were generated via use of CRISPR/Cas9 and Notch4 function was inhibited by an anti-Notch4 neutralizing antibody. The vasculature of postnatal retina from control animals was compared to either knockout or anti-Notch4 treated mice on measures of radial outgrowth, tip cell count, front density, capillary density, large vessel count, branching, and large vessel diameter. The experimental groups were then compared to each other to understand the degree to which pharmacologic inhibition recapitulates the genetic knockout phenotype. Our data shows a statistically significant reduction outgrowth and vein diameter in pharmacologic nulls and a trend towards increased tip cell density. This makes sense in the context of established literature and offers an explanation for why loss of Notch4 reduces tumor perfusion and growth, given that slower vessel growth and smaller vessels would impede blood flow and perfusion to tumors.
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The Effect of Genetic and Pharmacologic Loss of Notch4 on Angiogenesis
Notch signaling is critical to the formation of blood vessels, and although the role of Notch1 has been well characterized, less is known about Notch4 and the degree to which its function overlaps with Notch1. In order to quantify the effects of removing Notch4, Notch4 mutant mice were generated via use of CRISPR/Cas9 and Notch4 function was inhibited by an anti-Notch4 neutralizing antibody. The vasculature of postnatal retina from control animals was compared to either knockout or anti-Notch4 treated mice on measures of radial outgrowth, tip cell count, front density, capillary density, large vessel count, branching, and large vessel diameter. The experimental groups were then compared to each other to understand the degree to which pharmacologic inhibition recapitulates the genetic knockout phenotype. Our data shows a statistically significant reduction outgrowth and vein diameter in pharmacologic nulls and a trend towards increased tip cell density. This makes sense in the context of established literature and offers an explanation for why loss of Notch4 reduces tumor perfusion and growth, given that slower vessel growth and smaller vessels would impede blood flow and perfusion to tumors.