Description of the PhD thesis project
Research focus of
Our lab is broadly interested in membrane
trafficking and in its perturbations associated with human pathologies such as
cancer. We are elucidating the various biological and biophysical mechanisms of
the regulation of vesicular transport and membrane trafficking, using a
combination of approaches, including live cell imaging, micro-patterning and
microfabrication, reconstitution of transport events using model membranes and
intracellular microrheology with optical tweezers.
The lab is headed by Bruno Goud who is a world
leader in the field of RAB GTPases.
The thesis will be co-supervised by
Jean-Baptiste Manneville (biophysicist) and Stéphanie Miserey-Lenkei (cell
Abstract of the
Cells can sense and respond to external forces
and mechanotransduction events appear to be critical for most cellular
functions. While mechanotransduction has been extensively studied at the plasma
membrane and at the nucleus, the impact of forces on other organelles is still
Our PhD project will study mechanotransduction
at the Golgi apparatus (GA), a central organelle for intracellular transport
We will ask three questions:
1) Can external and internal forces
propagate to the GA and impact its tension?
2) Is the tension of the GA regulated
by actin dynamics and/or the composition of Golgi membranes and the Golgi
3) Do post-Golgi trafficking and
polarized secretion depend on the tension of the GA?
To achieve these goals, we will develop new
fluorescent probes to visualize and quantify tension at the GA. The probes will
be first tested in the mammalian RPE-1 cell line then used in the context of
pancreatic ductal adenocarcinoma (PDAC).
Our results should provide new
fundamental insights in the role played by mechanical tension in force
transduction at the level of the GA as well as a better understanding of the
physical mechanisms underlying polarized secretion during metastatic niche
formation in PDAC.
International, interdisciplinary &
intersectoral aspects of the project
The project will be developed in
collaboration with Laura Machesky (Beatson Institute, Glasgow, UK) expert in
PDAC metastatic niche formation, Sirio Dupont (U. Padova, Italy) expert in
YAP/TAZ signalling, and Aurélien Roux (U. Geneva, Switzerland) for the
development of Flipper tension probe specific to the GA.
We will collaborate with Aurélien Roux’s
industrial partner Spirochrome together with the chemistry group of Stefan
Matile (U. Geneva) to develop a Golgi tension probe.
The project mixes physico-chemical approaches
and tools (optical tweezers, microfluidics, force measurements, synthesis of
novel fluorescent tension sensors) with fundamental cell biology questions
related to mechanotransduction and to metastatic niche formation.
1) The Golgi apparatus and cell polarity:
Roles of the cytoskeleton, the Golgi matrix, and Golgi membranes. Ravichandran
Y, Goud B, Manneville JB. Curr Opin
Cell Biol. 2020 Feb; 62:104-113. doi: 10.1016/j.ceb.2019.10.003.
2) Role of a Kinesin Motor in Cancer Cell
Mechanics. Mandal K, Pogoda K, Nandi S, Mathieu S, Kasri A, Klein E, Radvanyi
F, Goud B, Janmey PA, Manneville JB.
Nano Lett. 2019 Nov 13; 19(11):7691-7702. doi: 10.1021/acs.nanolett.9b02592.
3) Extracellular matrix mechanical cues
regulate lipid metabolism through Lipin-1 and SREBP. Romani P, Brian I,
Santinon G, Pocaterra A, Audano M, Pedretti S, Mathieu S, Forcato M, Bicciato
S, Manneville JB, Mitro N, Dupont S.
Nat Cell Biol. 2019 Mar;21(3):338-347. doi: 10.1038/s41556-018-0270-5
4) RAB6 and microtubules restrict protein
secretion to focal adhesions. Fourriere L, Kasri A, Gareil N, Bardin S,
Bousquet H, Pereira D, Perez F, Goud B, Boncompain G, Miserey-Lenkei S. J Cell Biol. 2019 Jul 1;218(7):2215-2231.
5) Coupling fission and exit of RAB6 vesicles
at Golgi hotspots through kinesin-myosin interactions. Miserey-Lenkei S, Bousquet H, Pylypenko O, Bardin S, Dimitrov A,
Bressanelli G, Bonifay R, Fraisier V, Guillou C, Bougeret C, Houdusse A, Echard
A, Goud B. Nat Commun. 2017 Nov 1;8(1):1254. doi: 10.1038/s41467-017-01266-0.