MRes Bioengineering research project (new)
Title of the project:
Biophysics of Fenestrae Pore Formation
Supervisor 1:
Department: Dr. Darryl Overby
Imperial College campus: South Kensington
email: d.overby@imperial.ac.uk
Project Description: (up to 200 words)
Endothelial cells create a critical barrier to the transport of fluids and macromolecules between
compartments in the body, and alterations in endothelial transport contribute to disease.
Endothelial cells from several tissues have the ability to form small pores, less than 1 µm in
diameter, that allow passage of fluid or macromolecules across the endothelium. Disruption in
fenestrae/pore formation leads to elevated lipid content in the plasma, disrupted insulin transport
to the blood, and obstructed fluid transport within the eye. Thus, fenestrae/pore formation is
therefore fundamental for physiological homeostasis, but very little is known about the biophysics of
fenestra/pore formation.
Our laboratory has shown that pore formation is a mechano-sensitive process that is initiated by
biomechanical stretch[1] or transcellular pressure drop[2] applied to endothelial cells in culture. In
collaboration with investigators in Germany (Prof Ernst Tamm; U of Regensburg), we now have a
transgenic mouse that is deficient in a key protein (plasmalemma vesicle associated protein or
PLVAP) necessary for fenestra formation[3]. In this project, the student will isolate hepatic
endothelial cells from mice and examine how mechano-sensitive fenestra/pore formation is
disrupted by depletion of PLVAP. The student will also perform super-resolution microscopy to
examine the cytoarchitecture surrounding fenestrae and how fenestrae form dynamically following
stimulation with vascular endothelial growth factor. The findings from this project will have broad
applicability to understanding the role of mechanobiology and biophysics in regulating endothelial
transport in health and disease.
Key techniques: (please include only the names of techniques, not a description)
Cellular Biomechanics
Cell Culture and Molecular Biology
Work with Animals and Explant Cultures
Confocal and Electron Microscopy
Biofluid Mechanics
Control Engineering/Instrumentation for Experimentation
References: (up to 3 references related to the project)
1. Braakman ST, Pedrigi RM, Read AT, et al. Biomechanical strain as a trigger for
pore formation in Schlemm's canal endothelial cells. Exp Eye Res.
2014;127(C):224–235. doi:10.1016/j.exer.2014.08.003.
2.
Overby DR, Zhou EH, Vargas-Pinto R, et al. Altered mechanobiology of
Schlemm's canal endothelial cells in glaucoma. Proceedings of the National
Academy of Sciences. 2014;111(38):13876–13881. doi:10.1073/pnas.1410602111.
3.
Herrnberger L, Seitz R, Kuespert S, Bösl MR, Fuchshofer R, Tamm ER. Lack
of endothelial diaphragms in fenestrae and caveolae of mutant Plvap-deficient mice.
Histochem Cell Biol. 2012;138(5):709–724. doi:10.1007/s00418-012-0987-3.