MIGRATION, INVASION AND METASTASIS

www.beatson.gla.ac.uk/laura_machesky

Group Leader

Laura
Machesky
Research Scientists
Amelie Juin
Ben Tyrrell
Scientific Officer
Heather Spence
Clinical Research Fellows
Hayley Morris1
Richard Stevenson
Graduate Students
Loic Fort
Emma Woodham
Visitor
Emmy Borgmastars
1

MRC

Cancer metastasis requires cells to break away from the

primary tumour and to survive in a variety of environments
before settling into a new site. We aim to gain insights into
mechanisms of metastatic spread by determining the roles
of key actin cytoskeletal proteins, such as the actin filament
nucleation machinery and the bundling protein fascin, in
cancer cell invasive and migratory behaviour. The actin
cytoskeleton is important not only for cell strength and
migratory capacity but also for adhesion-dependent survival,
membrane trafficking and establishment of polarity. We aim
to understand how various actin regulators control these
processes and thus contribute to tumour initiation, growth
and spread as well as to fundamental mechanisms of
mammalian development.
Role of actin nucleating proteins in cell
migration, invasion and membrane trafficking
The Arp2/3 complex is the major inducer of
actin filaments in response to extracellular
signals. Members of the Wiskott-Aldrich
Syndrome Protein family (including WASP/NWASP, SCAR/WAVE, WASH, WHAMM and JMY)
transmit signals to the Arp2/3 complex to trigger
actin assembly. Each of these proteins is
regulated differently and one of our aims is to
understand the mechanisms of regulation and
the involvement of these proteins in invasion
and metastasis of cancer as well as their normal
cellular function. WASP family proteins regulate
actin assembly in multiple essential and
pathological cellular processes, such as
endocytic trafficking, protrusion of lamellipodia
and filopodia, cell division and assembly of

Figure 1
Mouse megakaryocyte labelled
with phalloidin (actin filaments,
red), ARPC2 antibody (Arp2/3
complex, green) and DAPI (DNA,
blue). The prominent dots are
podosomes. Photo by Hannah
Schachtner.

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SCIENTIFIC REPORT 2013 CANCER RESEARCH UK BEATSON INSTITUTE

invasive structures such as podosomes and

invadopodia.
Together with former postdoc Xinzi (Amber) Yu
and scientific officer Heather Spence and in
collaboration with Lynn McGarry and Emma
Shanks (RNAi Screening), we completed a
targeted screen for genes regulating invasion.
We continue to validate and study several new
targets involved in three-dimensional migration
and invasion. The most promising will be taken
forward into in vivo screens for invasion and
metastasis. Amber has moved on to a
postdoctoral position with Iain McNeish
(University of Glasgow).
This year, we (student Hannah Schachtner and
postdoc Simon Calaminus) published a study
showing that the platelet precursor cells,
megakaryocytes assemble actin-based
podosomes that resemble invadopodia of
cancer cells (Fig. 1 and Schachtner et al., Blood
2013; 121: 2542). Megakaryocyte podosomes
contain actin, Arp2/3 complex and WASP and
they are used for adhesion, spreading and
motility. Podosomes form an interconnected
network in cells that can sense and respond to
changes in the extracellular matrix environment.
We also found that megakaryocyte podosomes
could degrade matrix and protrude through

Sansom). We are interested to know whether the

role of N-WASP in invadopodia translates into
differences in tumour formation, progression
and spread in these models.

Figure 2
Expression of fascin in
a well-differentiated
mouse PDAC. Photo
shows immunofluorescence
staining of a murine tumour
expressing Pdx-1Cre KRasG12D
and p53R172H . Fascin is green,
E-cadherin is white, Slug is
red and DNA is blue. Photo
by Ang Li.

We are also studying the role of fascin-1 in

pancreatic ductal adenocarcinoma progression,
a project initiated by former student Ang Li (now
at Rockefeller University in New York) and now
continued by Amelie Juin, Hayley Morris and
students Emma Woodham and Loic Fort, and in
collaboration with Jen Morton and Owen
Sansom (Fig. 2). We found that fascin is a target
of the epithelial to mesenchymal transition in
pancreatic cancer and has a key role in early
tumour formation as well as metastatic
dissemination. Fascin mediates metastasis to the
peritoneal cavity via the formation of actin-rich
filopodia that enable cells to intercalate and
migrate through mesothelial cell layers and
seed a new tumour. Fascin also contributes to
liver metastasis. This work complements our
ongoing efforts to develop fascin inhibitor
compounds together with Martin Drysdale
(Drug Discovery).

Figure 3
Hindbrain immunostain
showing PECAM for blood
vessels (green), fascin (red)
and DNA (DAPI, blue) and
co-localisation in white of fascin
with endothelial cells. From Ma
et al., 2013. Photo by Ang Li.

native basement membranes. We speculate that

megakaryocytes may use podosomes for
extending proplatelet arms through the walls of
sinusoids or blood vessels to release platelets
into the bloodstream. In our review (Schachtner
et al., Cytoskeleton 2013; 70: 572), we discuss
podosomes as mechanosensors and mediators
of adhesion and matrix remodelling and the
relationship between podosomes and cancer
cell invadopodia. Hannah has taken up a
postdoc position with Tanja Maritzen in
Berlin, while Simon has moved on to a
lectureship in Hull.
We also continued our studies of the role
of WASH in actin networks of Dictyostelium
cells (together with Robert Insall, Park et al.,
Dev Cell 2013; 24: 169) and in breast cancer
cells. Postdoc Tobias Zech found a potential
connection between WASH and ERBB2
signalling, which we are studying further in
vitro and in vivo with the help of a pilot grant
from Breast Cancer Campaign. Tobias has
recently taken up a lectureship at the
University of Liverpool.
Role of actin regulatory proteins in
colorectal and pancreatic cancer
Clinical research fellow Richard Stevenson
identified a role of the actin bundling protein
fascin in tumour formation in colorectal cancer
models as well as a potential role in colitis. We
started new projects with MRC funded clinical
research fellow Hayley Morris on the role of
N-WASP in colorectal cancer (in collaboration
with Owen Sansom) and with postdoc Amelie
Juin on the role of N-WASP in pancreatic ductal
adenocarcinoma (with Jen Morton and Owen

Role of actin regulatory proteins in

melanoblast migration and melanoma
We also started new projects with Emma
Woodham and postdoc Ben Tyrrell on the role
of Rho family GTPases in melanocyte migration.
We previously showed that loss of Rac1 causes
major defects in melanoblast migration and
proliferation during development (Li et al., Dev
Cell 2011; 21: 722) and now we are studying the
roles of RhoA and Cdc42 in melanoblasts,
together with Cord Brakebusch (University of
Copenhagen, Denmark). With postdoc Yafeng
Ma, we published this year that loss of fascin
causes defects in melanoblast migration and
proliferation that are subtler than loss of Rac1
(Ma et al., Development 2013; 140: 2203). Fascin
is transiently expressed in murine melanoblasts
during their migration and is lost later when they
differentiate into melanocytes. Fascin is
expressed in some melanomas but is not tightly
correlated with stage or progression as it is in
many epithelial cancers.
Fascin is also present in tumour stroma and is
expressed by vascular endothelial cells, smooth
muscle cells and pericytes. Despite the
abundance of fascin in blood vessel walls, we
found that transplanted B16 melanomas
showed only slight differences in vascularisation
when the host was a fascin knockout. Tumour
growth was not affected by these small
differences, making fascin dispensable for
tumour angiogenesis. Likewise, developmental
angiogenesis does not seem to be greatly
affected by loss of fascin (Fig. 3 and Ma et al.,
Biol Open 2013; 2: 1187).
Publications listed on page 81

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MIGRATION, INVASION AND METASTASIS