Notch Regulates Vascular Collagen IV Basement Membrane Through Modulation of Lysyl Hydroxylase 3 Trafficking

<p>Collagen type IV (Col IV) is a basement membrane protein associated with early blood vessel morphogenesis and is essential for blood vessel stability. Defects in vascular Col IV deposition are the basis of heritable disorders, such as small vessel disease, marked by cerebral hemorrhage and drastically shorten lifespan. To date, little is known about how endothelial cells regulate the intracellular transport and selective secretion of Col IV in response to angiogenic cues, leaving a void in our understanding of this process. Our aim was to identify trafficking pathways that regulate Col IV deposition during angiogenic blood vessel development. We have identified the GTPase Rab10 as a major regulator of Col IV vesicular trafficking during vascular development using both <em>in vitro</em> imaging and biochemistry as well as in vivo models. Knockdown of Rab10 reduced <em>de novo</em> Col IV secretion <em>in vitro</em> and <em>in vivo</em>. Mechanistically, we determined that Rab10 is an indirect mediator of Col IV secretion partnering with atypical Rab25 to deliver the enzyme lysyl hydroxylase 3 (LH3) to Col IV-containing vesicles staged for secretion. Loss of Rab10 or Rab25 results in depletion of LH3 from Col IV-containing vesicles and rapid lysosomal degradation of Col IV. Furthermore, we demonstrate that Rab10 is Notch responsive, indicating a novel connection between permissive Notch-based blood vessel maturation programs and vesicle trafficking. Our results illustrate both a new trafficking-based component in the regulated secretion of Col IV and how this vesicle trafficking program interfaces with Notch signaling to fine-tune basement membrane secretion during blood vessel development.</p>

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Year dc:date.available

2021

Author dc:creator

• Gross, Stephen J.B.

Contributors dc:contributor

• Erich J. Kushner
• Yan Qin
• Bruce Appel
• Michelle Knowles

• Collagen IV
• Endothelial
• Lysyl hydroxylase 3
• Rab10
• Rab25
• Vasculature
• Biochemistry, Biophysics, and Structural Biology
• Biology
• Molecular Biology

Statement dc:rights

• <p>Copyright is held by the author. User is responsible for all copyright compliance.</p>

Language dc:language

en

Repository record dc:identifier

https://digitalcommons.du.edu/etd/1932

OAI identifier oai:identifier

oai:digitalcommons.du.edu:etd-2926