A Rac/Cdc42 exchange factor complex promotes formation of lateral filopodia and blood vessel lumen morphogenesis

Sabu Abraham; Margherita Scarcia; Richard D Bagshaw; Kathryn McMahon; Gary Grant; Tracey Harvey; Maggie Yeo; Filomena O G Esteves; Helene H Thygesen; Pamela F Jones; Valerie Speirs; Andrew M Hanby; Peter J Selby; Mihaela Lorger; T Neil Dear; Tony Pawson; Christopher J Marshall; Georgia Mavria

doi:10.1038/ncomms8286

A Rac/Cdc42 exchange factor complex promotes formation of lateral filopodia and blood vessel lumen morphogenesis

Sabu Abraham, Margherita Scarcia, Richard D Bagshaw, Kathryn McMahon, Gary Grant, Tracey Harvey, Maggie Yeo, Filomena O G Esteves, Helene H Thygesen, Pamela F Jones, Valerie Speirs, Andrew M Hanby, Peter J Selby, Mihaela Lorger, T Neil Dear, Tony Pawson, Christopher J Marshall, Georgia Mavria

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Abstract

During angiogenesis, Rho-GTPases influence endothelial cell migration and cell-cell adhesion; however it is not known whether they control formation of vessel lumens, which are essential for blood flow. Here, using an organotypic system that recapitulates distinct stages of VEGF-dependent angiogenesis, we show that lumen formation requires early cytoskeletal remodelling and lateral cell-cell contacts, mediated through the RAC1 guanine nucleotide exchange factor (GEF) DOCK4 (dedicator of cytokinesis 4). DOCK4 signalling is necessary for lateral filopodial protrusions and tubule remodelling prior to lumen formation, whereas proximal, tip filopodia persist in the absence of DOCK4. VEGF-dependent Rac activation via DOCK4 is necessary for CDC42 activation to signal filopodia formation and depends on the activation of RHOG through the RHOG GEF, SGEF. VEGF promotes interaction of DOCK4 with the CDC42 GEF DOCK9. These studies identify a novel Rho-family GTPase activation cascade for the formation of endothelial cell filopodial protrusions necessary for tubule remodelling, thereby influencing subsequent stages of lumen morphogenesis.

Original language	English
Article number	7286
Number of pages	14
Journal	Nature Communications
Volume	6
DOIs	https://doi.org/10.1038/ncomms8286
Publication status	Published - 1 Jul 2015

Keywords

Animals
Cytoskeleton
GTPase-Activating Proteins
Guanine Nucleotide Exchange Factors
Human Umbilical Vein Endothelial Cells
Humans
Mice, Inbred C57BL
Mice, Knockout
Neovascularization, Pathologic
Neovascularization, Physiologic
Pseudopodia
Vascular Endothelial Growth Factor A
cdc42 GTP-Binding Protein
rho GTP-Binding Proteins
Journal Article
Research Support, Non-U.S. Gov't

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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A RacCdc42 exchange factor complex promotes formation of lateral filopodia and blood vessel lumen morphogenesis
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Final published version, 4.62 MBLicence: CC BY

Valerie Speirs
- School of Medicine, Medical Sciences & Nutrition, Medical Sciences - Chair in Molecular Oncology
- Institute of Medical Sciences
Person: Academic

Cite this

Abraham, S., Scarcia, M., Bagshaw, R. D., McMahon, K., Grant, G., Harvey, T., Yeo, M., Esteves, F. O. G., Thygesen, H. H., Jones, P. F., Speirs, V., Hanby, A. M., Selby, P. J., Lorger, M., Dear, T. N., Pawson, T., Marshall, C. J., & Mavria, G. (2015). A Rac/Cdc42 exchange factor complex promotes formation of lateral filopodia and blood vessel lumen morphogenesis. Nature Communications, 6, [7286]. https://doi.org/10.1038/ncomms8286

Abraham, S, Scarcia, M, Bagshaw, RD, McMahon, K, Grant, G, Harvey, T, Yeo, M, Esteves, FOG, Thygesen, HH, Jones, PF, Speirs, V, Hanby, AM, Selby, PJ, Lorger, M, Dear, TN, Pawson, T, Marshall, CJ & Mavria, G 2015, 'A Rac/Cdc42 exchange factor complex promotes formation of lateral filopodia and blood vessel lumen morphogenesis', Nature Communications, vol. 6, 7286. https://doi.org/10.1038/ncomms8286

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title = "A Rac/Cdc42 exchange factor complex promotes formation of lateral filopodia and blood vessel lumen morphogenesis",

abstract = "During angiogenesis, Rho-GTPases influence endothelial cell migration and cell-cell adhesion; however it is not known whether they control formation of vessel lumens, which are essential for blood flow. Here, using an organotypic system that recapitulates distinct stages of VEGF-dependent angiogenesis, we show that lumen formation requires early cytoskeletal remodelling and lateral cell-cell contacts, mediated through the RAC1 guanine nucleotide exchange factor (GEF) DOCK4 (dedicator of cytokinesis 4). DOCK4 signalling is necessary for lateral filopodial protrusions and tubule remodelling prior to lumen formation, whereas proximal, tip filopodia persist in the absence of DOCK4. VEGF-dependent Rac activation via DOCK4 is necessary for CDC42 activation to signal filopodia formation and depends on the activation of RHOG through the RHOG GEF, SGEF. VEGF promotes interaction of DOCK4 with the CDC42 GEF DOCK9. These studies identify a novel Rho-family GTPase activation cascade for the formation of endothelial cell filopodial protrusions necessary for tubule remodelling, thereby influencing subsequent stages of lumen morphogenesis. ",

keywords = "Animals, Cytoskeleton, GTPase-Activating Proteins, Guanine Nucleotide Exchange Factors, Human Umbilical Vein Endothelial Cells, Humans, Mice, Inbred C57BL, Mice, Knockout, Neovascularization, Pathologic, Neovascularization, Physiologic, Pseudopodia, Vascular Endothelial Growth Factor A, cdc42 GTP-Binding Protein, rho GTP-Binding Proteins, Journal Article, Research Support, Non-U.S. Gov't",

author = "Sabu Abraham and Margherita Scarcia and Bagshaw, {Richard D} and Kathryn McMahon and Gary Grant and Tracey Harvey and Maggie Yeo and Esteves, {Filomena O G} and Thygesen, {Helene H} and Jones, {Pamela F} and Valerie Speirs and Hanby, {Andrew M} and Selby, {Peter J} and Mihaela Lorger and Dear, {T Neil} and Tony Pawson and Marshall, {Christopher J} and Georgia Mavria",

note = "We thank Hironori Katoh for EGFP DOCK4; Vijay Yajnik for flag-tagged DOCK4 and deletion mutants; Martin Schwartz for the DOCK9 expression construct; Len Stephens for RhoG expression construct; Jean-Fran{\c c}ois C{\^o}t{\'e} for GST–ELMO. We thank Janine Erler for advice on CA-IX staining and Kiran Kulkarni for discussions on DOCK9; Demelza Bird and Kris Wronski for the tumour experiments; Ben Woodman and staff at St James{\textquoteright} Biological Services for ES microinjections and animal husbandry; Biological Services Unit, ICR, for animal husbandry; Mike Shires for optimization of DOCK4 IHC; Anne Sanford and Rebecca Horsfall for technical assistance; Sean Lawler and other colleagues for commenting on the manuscript. The work was funded through project grant from Cancer Research UK (C107/A7631) awarded to C.J.M. and G.M.; project grant from Yorkshire Cancer Research (L357) and pilot grant from Breast Cancer Campaign (2012MaySP047) awarded to G.M. T.P. was supported by grants from the Ontario Research Fund GL2 program and CIHR (MOP-6849); and R.D.B. was supported by a CIHR fellowship. C.J.M. is a Gibb Life Fellow of Cancer Research UK. ",

year = "2015",

month = jul,

day = "1",

doi = "10.1038/ncomms8286",

language = "English",

volume = "6",

journal = "Nature Communications",

issn = "2041-1723",

publisher = "Nature Publishing Group",

}

TY - JOUR

T1 - A Rac/Cdc42 exchange factor complex promotes formation of lateral filopodia and blood vessel lumen morphogenesis

AU - Abraham, Sabu

AU - Scarcia, Margherita

AU - Bagshaw, Richard D

AU - McMahon, Kathryn

AU - Grant, Gary

AU - Harvey, Tracey

AU - Yeo, Maggie

AU - Esteves, Filomena O G

AU - Thygesen, Helene H

AU - Jones, Pamela F

AU - Speirs, Valerie

AU - Hanby, Andrew M

AU - Selby, Peter J

AU - Lorger, Mihaela

AU - Dear, T Neil

AU - Pawson, Tony

AU - Marshall, Christopher J

AU - Mavria, Georgia

N1 - We thank Hironori Katoh for EGFP DOCK4; Vijay Yajnik for flag-tagged DOCK4 and deletion mutants; Martin Schwartz for the DOCK9 expression construct; Len Stephens for RhoG expression construct; Jean-François Côté for GST–ELMO. We thank Janine Erler for advice on CA-IX staining and Kiran Kulkarni for discussions on DOCK9; Demelza Bird and Kris Wronski for the tumour experiments; Ben Woodman and staff at St James’ Biological Services for ES microinjections and animal husbandry; Biological Services Unit, ICR, for animal husbandry; Mike Shires for optimization of DOCK4 IHC; Anne Sanford and Rebecca Horsfall for technical assistance; Sean Lawler and other colleagues for commenting on the manuscript. The work was funded through project grant from Cancer Research UK (C107/A7631) awarded to C.J.M. and G.M.; project grant from Yorkshire Cancer Research (L357) and pilot grant from Breast Cancer Campaign (2012MaySP047) awarded to G.M. T.P. was supported by grants from the Ontario Research Fund GL2 program and CIHR (MOP-6849); and R.D.B. was supported by a CIHR fellowship. C.J.M. is a Gibb Life Fellow of Cancer Research UK.

PY - 2015/7/1

Y1 - 2015/7/1

N2 - During angiogenesis, Rho-GTPases influence endothelial cell migration and cell-cell adhesion; however it is not known whether they control formation of vessel lumens, which are essential for blood flow. Here, using an organotypic system that recapitulates distinct stages of VEGF-dependent angiogenesis, we show that lumen formation requires early cytoskeletal remodelling and lateral cell-cell contacts, mediated through the RAC1 guanine nucleotide exchange factor (GEF) DOCK4 (dedicator of cytokinesis 4). DOCK4 signalling is necessary for lateral filopodial protrusions and tubule remodelling prior to lumen formation, whereas proximal, tip filopodia persist in the absence of DOCK4. VEGF-dependent Rac activation via DOCK4 is necessary for CDC42 activation to signal filopodia formation and depends on the activation of RHOG through the RHOG GEF, SGEF. VEGF promotes interaction of DOCK4 with the CDC42 GEF DOCK9. These studies identify a novel Rho-family GTPase activation cascade for the formation of endothelial cell filopodial protrusions necessary for tubule remodelling, thereby influencing subsequent stages of lumen morphogenesis.

AB - During angiogenesis, Rho-GTPases influence endothelial cell migration and cell-cell adhesion; however it is not known whether they control formation of vessel lumens, which are essential for blood flow. Here, using an organotypic system that recapitulates distinct stages of VEGF-dependent angiogenesis, we show that lumen formation requires early cytoskeletal remodelling and lateral cell-cell contacts, mediated through the RAC1 guanine nucleotide exchange factor (GEF) DOCK4 (dedicator of cytokinesis 4). DOCK4 signalling is necessary for lateral filopodial protrusions and tubule remodelling prior to lumen formation, whereas proximal, tip filopodia persist in the absence of DOCK4. VEGF-dependent Rac activation via DOCK4 is necessary for CDC42 activation to signal filopodia formation and depends on the activation of RHOG through the RHOG GEF, SGEF. VEGF promotes interaction of DOCK4 with the CDC42 GEF DOCK9. These studies identify a novel Rho-family GTPase activation cascade for the formation of endothelial cell filopodial protrusions necessary for tubule remodelling, thereby influencing subsequent stages of lumen morphogenesis.

KW - Animals

KW - Cytoskeleton

KW - GTPase-Activating Proteins

KW - Guanine Nucleotide Exchange Factors

KW - Human Umbilical Vein Endothelial Cells

KW - Humans

KW - Mice, Inbred C57BL

KW - Mice, Knockout

KW - Neovascularization, Pathologic

KW - Neovascularization, Physiologic

KW - Pseudopodia

KW - Vascular Endothelial Growth Factor A

KW - cdc42 GTP-Binding Protein

KW - rho GTP-Binding Proteins

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1038/ncomms8286

DO - 10.1038/ncomms8286

M3 - Article

C2 - 26129894

VL - 6

JO - Nature Communications

JF - Nature Communications

SN - 2041-1723

M1 - 7286

ER -

A Rac/Cdc42 exchange factor complex promotes formation of lateral filopodia and blood vessel lumen morphogenesis

Abstract

Keywords

UN SDGs

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