Determining the role of Protocadherin-1 in angiogenesis

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Vecchiato, Werther (2022). Determining the role of Protocadherin-1 in angiogenesis. University of Birmingham. Ph.D.

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Abstract

Protocadherin-1 (PCDH1) is a single-pass transmembrane protein belonging to the Cadherin superfamily of adhesion molecules. Protocadherins are involved in neuronal guidance, however a biological role for PCDH1 has not been established. Many neuronal guidance cues are co-opted as angiogenic cues, and PCDH1 is expressed in both neural tissue and the endothelium. The hypothesis explored in this thesis is that PCDH1 participates in angiogenesis. Different siRNA duplexes reduced PCDH1 expression in human umbilical vein endothelial cells (HUVEC), thereby impairing chemotaxis and connectivity on Matrigel. However, these phenotypes were abolished when all duplexes were delivered as part of a siRNA-mixture. Transcriptomic analysis of siRNA-mixture-treated cells suggested that endogenous PCDH1 has a modestly anti-angiogenic role, but identified no clear candidate to mediate the phenotypes observed initially. Mass spectrometric analysis of the PCDH1 intracellular interactome detected Desmoplakin, a cytosolic scaffold protein. Proximity-sensitive immunofluorescence experiments corroborated these findings, and biochemical analysis revealed that the first four Spectrin repeats of Desmoplakin co-purify with PCDH1. The truncated, soluble PCDH1 ectodomain did not alter HUVEC connectivity on Matrigel, and Pcdh1-knockout embryos displayed no vascular abnormalities at E13.5. Overall, these findings indicate that PCDH1 does not play a critical role in angiogenesis but they do identify a novel intracellular interacting partner for PCDH1.

Type of Work:

Thesis (Doctorates > Ph.D.)

Award Type:

Doctorates > Ph.D.

Supervisor(s):