MOLECULAR BIOLOGY OF THE CELL - PROGRAM 2019-2020
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Pasteur course
MOLECULAR BIOLOGY OF THE CELL
PROGRAM 2019-2020
JANUARY 13 – FEBRUARY 13, 2020MOLECULAR BIOLOGY OF THE CELL
2019 - 2020
January, 13 - February, 13(*), 2020
(*) exams included
Directors of the course
Chiara ZURZOLO
Institut Pasteur
Paris, France
Roberto BRUZZONE Philippe CHAVRIER
HKU – Pasteur Research Pole Institut Curie – Research Section
The University of Hong Kong, Hong Kong SAR Paris, France
Location
Centre d'Enseignement de l'Institut Pasteur
Lectures : Pavillon Louis Martin, building 09, Room n°1
Practicals : Pavillon Louis Martin, building 09, 1st floor
28, rue du Docteur Roux
75724 Paris Cedex 15
(*) for Non-Master students, the course will end on FEBRUARY 7, 2020 at 6:00PM.Description of the course The Molecular Biology of the Cell course is an intensive laboratory and lecture course of five weeks divided into weekly modules, each focusing on a cutting-edge aspect of cell biology. It is composed of lectures given by internationally renowned scientists, and of two practical sessions organized together with teams from the Curie and the Pasteur Institutes. The main topics of the course extend across the cell biology of infection, cancer, signaling, epigenetics and intracellular trafficking, emphasizing new experimental approaches. The availability of the core Imaging Platform at Institut Pasteur will introduce students to advanced techniques for the dynamic visualization of cells in health and disease. Participants are selected from Master 2 students of the University of Paris 6, Paris 7 and Paris XI and foreign postgraduate programs. The course is intended to be a platform of excellence in which students can meet and closely interact with worldwide top-level scientists to discuss, exchange ideas and establish valuable contacts in the perspective of establishing a network of young cell biologists at an early stage in their careers. Students will be able to understand the importance of basic research and of a broad interdisciplinary approach to improve human health. We also expect to provide orientations and mentoring to help course alumni in their future career. The 2019-2020 course is subdivided into two modules. The first module will provide an overview of key biological concepts required to understand processes that regulate the dynamics of proteins and lipids at the plasma membrane. Topics of lectures include: mechanical forces and caveolae- dependent signaling; glycosylation, endocytosis, and intracellular sorting. It will be complemented by a practicum during which students will investigate caveolae dynamics and mechanotransduction and carbohydrate-based mechanisms to build endocytic pits using chemical biology and imaging techniques. The second module will review key biological concepts required to understand tissue morphogenesis. Topics of lectures include: mechanical forces and morphogenesis; epithelial organization and apico-basal cell polarity; collective migration and cell-cell adhesion; micropattern and self organisation; mechanotransduction. The practicum will discuss the main parameters regulating the formation and the remodeling of multicellular tubes in vitro, using micropatterns and microfluidics, and in vivo, using the formation of the tracheal system in Drosophila and the formation of the vasculature and the neural tube during Zebrafish development.
Practicum 1
ENDOCYTOSIS, SIGNALING, CAVEOLAE & GLYCOSYLATION
In the first part of this practicum, we will study how cells use caveolae to translate physical stimuli
into biochemical signals by mechanotransduction by which information from the cell surface is
transmitted to the nucleus, where gene expression is regulated. Mechanotransduction controls
multiple cellular aspects including, but not limited to, cell growth, shape, or differentiation.
Abnormal cell responses to external and internal mechanical constraints are often associated with
human pathologies such as heart diseases, myopathies and cancer. Caveolae are 60-80 nm bulb-
like plasma membrane invaginations discovered more than sixty years ago. Caveolae are
generated through tight association of caveolin oligomers, its main structural component, and are
stabilized by the assembly of cytoplasmic cavins into a coat-like structure around the caveolae
bulb. Our laboratory has shown that caveolae can act as mechanosensing and mechanoprotection
cellular organelles. Under increase of membrane tension generated by various types of
mechaniocal stress such as cell swelling or stretching, caveolae flatten out immediately to provide
additional surface area and prevent the rupture of the plasma membrane. The central role of
caveolae in cell mechanics has been extended to a large number of cell types.
During the first week, we will:
1) monitor the cytosolic release of the EHD2 ATPase from mechanically disassembled
caveolae and its subsequent translocation to the nucleus to mediate mechanotransduction.
2) monitor EHD2 SUMOylation, a post translational modification induced by mechanical
stress and involved in EHD2 nucleocytoplasmic shuttling.
Through the use of microscopy techniques, this practicum aims at illustrating the key role of
caveolae in the response to mechanical stress. It will focus on EHD2, which, by combining both
mechanosensing and mechanotransducing activities, plays a central role in the mechanical cell
response mediated by caveolae.
In the second part of the practicum, we will study how given endocytic pathways may be early
signatures for subsequent intracellular compartmentalization of cargo proteins. We will focus on
the well-characterized clathrin pathway as well as on a new endocytic process that was recently
hypothesized by our lab, termed the Glycolipid-Lectin (GL-Lect) mechanism. In this latter,
glycosylated cargoes, secreted galectins, and glycosphingolipids are key components to initiate
membrane curvature for subsequent clathrin-independent endocytic carriers’ formation.
For this practical course, we will dissect the endocytic behavior of a particular cargo, the adhesion
protein α5β1 integrin, depending on its conformational state. Using the chemical BG/SNAP-tag tool,
we have recently demonstrated that the non-ligand-bound conformation of β 1 integrin undergoes
retrograde trafficking from the plasma membrane to the Golgi compartment. Alteration of canonical
retrograde transport machinery is strongly linked with a defect in cell adhesion as well as a dramatic
loss of persistent cell migration.
We will first at all obtain a proof of concept demonstration for the use of the BG/SNAP tool for the
study of retrograde trafficking, using the receptor-binding B-subunit of Shiga toxin (STxB) as a
model cargo. Its Golgi accumulation will be monitored by microscopy techniques. We will then
analyze by biochemical methods the specific retrograde transport of the different conformations of
β1 integrin using conformational state-specific antibodies, in different drug treatment conditions(EGF, GENZ, CBD). In the second part of the program, we will focus on endocytosis. Using
fluorescence microscopy and imaging tools, we will analyze the contribution of the GL-Lect
mechanism to the endocytic uptake of β1 integrin in different conformational states. Together, our
program will provide insights on how the entry mode of a cargo represents a signature for its
intracellular compartmentalization and biological activity.
Practicum 2
ALL YOU WANT TO KNOW ABOUT TUBE FORMATION
Tissue morphogenesis is a multiscale problem integrating modification of cell shape, cell migration,
cell position swapping, remodeling of the extracellular environment as well as large scale
coordination of cell behavior at the tissue level. To illustrate the different approaches that can be
used to tackle a standard morphogenesis problem, we will use a combination of model systems to
characterize the main parameters regulating the formation and remodeling of multicellular tubes, a
common tissue shape present throughout Metazoans. Three independent examples of tube
formation will be studied by every student in subgroups:
1. The formation of a tube in vitro: We will study the parameters affecting the spontaneous
formation of tubes in vitro using a combination of endothelial cells and fibroblasts through
long term live imaging, micropatterns and microfluidics.
2. Morphogenesis of multicellular tubes in vivo: We will study the formation of the
intersegmental vessel and the neural tube in Zebrafish embryos. By combining knockdown
experiment and live imaging we will characterize cellular movements and deformations
during lumen cavitation and tube elongation. We will also introduce general concepts
regarding Zebrafish handling and live imaging of embryos.
3. Remodeling tubes in vivo: We will study the morphogenesis of the tracheal system in
Drosophila embryos. Combining fast live imaging, cell tracking and laser perturbation
approaches, we will test which type of forces contribute to the elongation of the terminal
dorsal branches of the tracheas. General concepts regarding Drosophila development, fly
handling as well as live imaging will also be introduced.
During the second week, data collected on those systems will be analyzed quantitatively using
various software and custom procedures (Matlab, Fiji, Imaris). The results obtained on each system
will be discussed and compared to outline the universal features regulating tube
formation/remodeling. This module will illustrate how the combination of several model systems,
with different technical advantages and limitations, can help to answer a similar biological question.MOLECULAR BIOLOGY OF THE CELL 2019-2020
WEEK 1 January 13-17, 2020
MODULE 1: ENDOCYTOSIS, SIGNALING, CAVEOLAE & GLYCOSYLATION
WEEK1: Christine VIARIS, Cédric BLOUIN and Christophe LAMAZE, Institut Curie
Monday, January 13
9:00 – 9:30 Introduction & Presentation of the course Roberto BRUZZONE
(HKU-Pasteur Research Pole)
Philippe CHAVRIER
(Institut Curie)
Chiara ZURZOLO
(Institut Pasteur)
9:30 – 10:00 Administrative issues &
Presentation of the Education Center Education Center
(Institut Pasteur)
10:00 – 10:30 Photos for badges (front desk, 25 rue du Dr Roux)
10:30 – 11:30 Self-presentation of the Students (5 min max, 2 slides each,
background + project)
12:00 – 13:00 BCI departmental seminar (Amphi JACOB) Serge MOSTOWY
"Septins and cell-autonomous immunity" (Imperial College, London, UK)
14:30 – 16:30 TBA Serge MOSTOWY
(Imperial College, London, UK)
17:00 – 18:00 Self-presentation of the Students
(5 min max, 2 slides each, background + project)
18:00 – 18:30 Presentation of practical sessions and exam
Tuesday, January 14
09:00 – 12:00 Practicum 1 – Day 1 C.VIARIS, C. BLOUIN, C. LAMAZE
Introduction Practicum
13:30 – 15:30 Control of signal transduction by caveolae Christophe LAMAZE
mechanics (Institut Curie)
13:30 – 18:00 Practicum 1 – Day 1
18:30 – 19:30 Welcome party (Cafétéria, ground floor, Education Center)Wednesday, January 15
9:00 – 11:00 Cell biology of neural progenitors and neural stem cells Laure BALLY-CUIF
(Institut Pasteur)
11:30 – 12:30 Practicum 1 – Day 2
Experiments
13:30 – 18:00 Practicum 1 – Day 2 (continued)
Experiments
Thursday, January 16
9:00 – 11:00 Integrins in cancer progression Johanna IVASKA
(University of Turku, Finland)
11:30 – 18:00 Practicum 1 – Day 3
Experiments
Friday, January 17
9:30 – 12:00 Fine-tuning killer cell secretion: single cell Gillian GRIFFITHS
and super-resolution approaches (Institute for Medical Research, Cambridge, UK)
12:15 – 12:30 Group picture
13:30 – 16:00 Practicum 1 – Day 4
Image acquisition
16:00 – 18:00 Data AnalysisMOLECULAR BIOLOGY OF THE CELL 2019-20120
WEEK 2 JANUARY 20-24, 2020
MODULE 1: ENDOCYTOSIS, SIGNALING, CAVEOLAE & GLYCOSYLATION
WEEK2: Massiullah SHAFAQ-ZADAH and Estelle DRANSART, Institut Curie
Monday, January 20
9:00 – 12:00 Practicum 1 – Day 5 Massiullah SHAFAQ-ZADAH, Estelle DRANSART
Introduction and experiments (Institut Curie)
12:00 – 13:00 BCI departmental seminar (Amphi JACOB) Fernando Martin BELMONTE
Organ tube formation in vertebrates:
a tale of autophagy and shape control (CBM Severo Ochoa, Madrid, Spain)
14:30 – 16:30 Mechanisms for the acquisition of cell Fernando Martin BELMONTE
polarity during epithelial morphogenesis (CBM Severo Ochoa, Madrid, Spain)
16:30 – 18:00 Practicum 1 – Day 5 Massiullah SHAFAQ-ZADAH, Estelle DRANSART
Experiments (continued) (Institut Curie)
Tuesday, January 21
9:00 – 12:00 Practicum 1 – Day 6
Experiments
13:30 – 15:30 Glycosphingolipid-dependent and lectin-driven Ludger JOHANNES
construction of endocytic pits (Institut Curie)
15:30 – 18:00 Practicum 1 – Day 6
Experiments (continued)
Wednesday, January 22
9:00 – 12:00 Practicum 1 – Day 7
Experiments
13:30 – 15:30 Dynamics of lipid membranes and protein coats Bruno ANTONNY
(IPMC, Valbonne, France)
15:30 – 18:00 Practicum 1 – Day 7
Finish experiments, observations and data analysisThursday, January 23
9:30 – 12:00 The complexity and the simplicity Maria MOTA
of host-Plasmodium interactions (Instituto de Medicina Molecular, Lisboa, Portugal)
13:30 – 16:30 Practicum 1 – Day 8
End of experiments, observations and data analysis
16:30 – 18:00 Preparation of presentations
Friday, January 24
9:30 – 12:00 Endocytosis, signaling and cancer Pier Paolo DI FIORE
(Instituto Europeo di Oncologia, Milano, Italy)
16:30 – 18:00 PresentationsMOLECULAR BIOLOGY OF THE CELL 2019-2020
WEEK 3 JANUARY 27-31, 2020
MODULE 2: How to Build a tube
Romain LEVAYER, Daria BONAZZI, Dorian OBINO, Nicolas DRAY, Institut Pasteur,
and Manuel THÉRY, CEA
Monday, January 27
9:30 – 11:30 Morphogenesis: a noisy path to order Buzz BAUM
(University College, London, UK)
12:00 – 13:00 BCI departmental seminar (Amphi JACOB) Buzz BAUM
Cancer cell division (University College, London, UK
14:30 – 18:00 Practicum 2 – Day 1 General introduction tube morphogenesis and preparation of
the slides for micropattern
Tuesday, January 28
9:00 – 11:00 From spindle orientation to tissue morphogenesis Yohanns BELLAÏCHE
(Institut Curie)
11:30 – 13:00 Cytoskeleton self-organization Manuel THERY
(CEA, Hôpital Saint-Louis, France)
14:30 – 18:00 Experiments with Drosophila (group1) and Zebrafish (group 2)
Wednesday, January 29
9:00 – 10:30 TBA Daria BONAZZI
(Institut Pasteur)
11:00 – 13:00 Fine tuning of tissue morphogenesis by cell death: Romain LEVAYER
the role of cell competition in development (Institut Pasteur)
and disease
14:30 – 18:00 Practicum 2 – Day 2
Experiments with Drosophila (group2) and Zebrafish (group 1)
Thursday, January 30
9:00 – 18:00 Practicum 2 – Day 3
Micropattern and microfluidics experimentsFriday, January 31
9:00 – 18:00 Practicum 2 – Day 4
Micropattern and microfluidics experimentsMOLECULAR BIOLOGY OF THE CELL 2019-2020
WEEK 4 FEBRUARY 3-7, 2020
MODULE 2: How to Build a tube
Romain LEVAYER, Daria BONAZZI, Dorian OBINO, Nicolas DRAY, Institut Pasteur,
and Manuel THÉRY, CEA
Monday, February 3
9:00 – 12:00 Practicum 2 – Day 5
Imaging of microfluidic results
12:00 – 13:00 BCI departmental seminar (Amphi JACOB) Markus AFFOLTER
From the genome to the proteome: novel approaches to study
the proteome in vivo (Biozentrum, Basel, Switzerland)
14:30 – 16:30 Forming tubes in drosophila and zebrafish: from Markus AFFOLTER
the trachea to the vasculature (Biozentrum, Basel, Switzerland)
Tuesday, February 4
09:30 – 12:00 SNARE: intracellular membrane fusion and much more Thierry GALLI
(Institute of Psychiatry and Neuroscience, Paris)
13:30 – 17:30 Practicum 2 – Day 6
Image acquisition of the micropattern
Wednesday, February 5
9:30 – 11:30 Mechanics of blastocyst morphogenesis Jean-Leon MAÎTRE
(Institut Curie)
13:30 – 17:30 Practicum 2 – Day 7
Data analysis Drosophila and Zebrafish movies
Thursday, February 6
9:30 – 12:00 Cytoskeletal crosstalk during Sandrine ETIENNE-MANNEVILLE
cell migration (Institut Pasteur)
13:30 – 17h30 Preparation of presentationsFriday, February 7
9:30 – 12 :00 Tunneling nanotubes: a novel mechanism Chiara ZURZOLO
of intercellular communication in physiology and pathology (Institut Pasteur)
13:00 – 16:00 Presentations
17:00 – 18:30 Oral Examination for Non-Master studentsMOLECULAR BIOLOGY OF THE CELL 2019-2020
WEEK 5 FEBRUARY 10-13, 2020
FINAL EXAMINATION
Monday, February 10 to Tuesday, February 11
Project preparation.
The written project has to be submitted to the Course Committee by Tuesday night.
Wednesday, February 12
Preparation of the oral examination
Thursday, February 13
9:00 – 18:00 Oral examinationDETAILED DESCRIPTION OF THE EXAMINATION
Oral examination on Thursday 13 February, 2020 (mark on a 1-20 scale, coefficient 1):
Critical analysis of a scientific article and presentation of an imaginary 3-year research project as
follow-up of the results of the article.
Presentation: 13 minutes; questions: 7 minutes; total duration: 20 minutes
Organization of the oral presentation:
The presentation is open to the public
Slides (Powerpoint or other supported formats)
The scientific articles will be given to students during the first week of the course. Each student will
write a fictional project intended to be a follow-up of the article received and submit a 4/5 page
document to the members of the jury no later than Tuesday 9th February at 20:00. This document
should include:
Summary of the article (max 1 page)
Aims and description of the project (max 3 pages including figures if appropriate)
References (max 1 page), using the style of a cell biology journal (e.g., JCB, JCS, MBC, Cell,
NCB….)
DESCRIPTION DETAILEE DE L’EXAMEN
Examen oral le jeudi 13 février 2020 (note sur 20, coefficient 1) :
Présentation critique d’un article et discussion d’un projet fictif sur 3 ans découlant de ces résultats.
Présentation : 13 minutes ; questions du jury : 7 minute ; durée totale : 20 minutes.
Organisation de la présentation orale :
Exposé public de chaque étudiant devant le jury
Diapositives (logiciel Powerpoint ou autre format compatible)
Les articles scientifiques seront donnés aux étudiants pendant la première semaine de cours. Le
projet fictif est présenté dans un document de 4/5 pages à remettre au jury au plus tard le mardi 9
février à 20h00, comprenant :
Résumé de l’article (max 1 page)
Objectifs et description du projet (max 3 pages, figures incluses)
Bibliographie (max 1 page) selon le style d’un journal type JCB, JCS, MBC, Cell, NCB…MOLECULAR BIOLOGY OF THE CELL 2019-2020
ADDRESS DETAILS
DIRECTORS OF THE COURSE
Chiara ZURZOLO
Membrane Trafficking & Pathogenesis Unit
Institut Pasteur
28, rue du Dr Roux
75724 Paris Cedex 15, France
Tel +33 - (0)1 45 68 82 77
chiara.zurzolo@pasteur.fr
Roberto BRUZZONE Philippe CHAVRIER
HKU - Pasteur Research Pole Institut Curie - Research Section
The University of Hong Kong CNRS UMR 144
5 Sassoon Road 26, rue d’Ulm
Pokfulam, Hong Kong SAR 75248 Paris Cedex 05, France
Tel +852 - 2831 5522 Tel +33 - (0)1 56 24 63 59
bruzzone@hku.hk philippe.chavrier@curie.frLECTURERS Mr AFFOLTER Markus markus.affolter@unibas.ch Biozentrum University of Basel Basel, Switzerland Mr ANTONNY Bruno antonny@ipmc.cnrs.fr Dynamique des membranes et manteaux protéiques IPMC Valbonne, France Ms BALLY-CUIF Laure laure.bally-cuif@pasteur.fr Department of Developmental and Stem Cell Biology Institut Pasteur Paris, France Mr BAUM Buzz b.baum@ucl.ac.uk MRC Laboratory for Molecular Biology University College London, UK Mr BELLAICHE Yohanns Yohanns.Bellaiche@curie.fr Genetics and Developmental Biology Institut Curie Paris, France Mr BELMONTE Fernando Martin fmartin@cbm.csic.es Department of Development and Differentiation Centro de Biologia Molecular Severo Ochoa Madrid, Spain Ms BONAZZI Daria daria.bonazzi@pasteur.fr Department of Cell Biology and Infection Institut Pasteur Paris, France Mr CHAVRIER Philippe Philippe.Chavrier@curie.fr Membrane and Cytoskeleton Dynamics Institut Curie Paris, France Ms ETIENNE-MANNEVILLE Sandrine Sandrine.etienne-manneville@pasteur.fr Department of Cell Biology and Infection Institut Pasteur Paris, France
Mr DI FIORE Pier Paolo pierpaolo.difiore@ieo.it Molecular Carcinogenesis and Stem Cell Biology Istituto Europeo di Oncologia Milano, Italy Mr GALLI Thierry Thierry.galli@inserm.fr Inserm U894 Insitute of Psychiatry and Neuroscience Paris, France Ms GRIFFITHS Gillian gg305@cam.ac.uk Cambridge Institute for Medical Research Department of Medicine, University of Cambridge Cambridge, UK Ms IVASKA Johanna joivaska@utu.fi Cell Adhesion and Cancer Laboratory Turku Centre for Biotechnology, University of Turku Turku, Finland Mr JOHANNES Ludger Ludger.Johannes@curie.fr Cellular and Chemical Biology/Endocytic Trafficking and Intracellular Delivery Institut Curie Paris, France Mr LAMAZE Christophe Christophe.lamaze@curie.fr Cellular and Chemical Biology/Membrane Mechanics and Dynamics of Intracellular Signaling Institut Curie Paris, France Mr LEVAYER Romain romain.levayer@pasteur.fr Department of Developmental and Stem Cell Biology Institut Pasteur Paris, France Mr MAITRE Jean-Leon Jean-Leon.Maitre@curie.fr Genetics and Developmental Biology/Mechanics of Mammalian Development Institut Curie Paris, France Mr MOSTOWY Serge s.mostowy@imperial.ac.uk Department of Infectious Diseases Imperial College London, UK Ms MOTA Maria mmota@medicina.ulisboa.pt Instituto de Medicina Molecular University of Lisboa Lisboa, Portugal
Mr THERY Manuel manuel.thery@cea.fr
Physics of Cytoskeleton & Morphogenesis
CEA, Hôpital Saint-Louis
Paris, France
PRACTICUM
Mr BLOUIN Cédric Cedric.Blouin@curie.fr
Cellular and Chemical Biology/Membrane Mechanics and Dynamics of Intracellular Signaling
Institut Curie
Paris, France
Ms BONAZZI Daria daria.bonazzi@pasteur.fr
Department of Cell Biology and Infection
Institut Pasteur
Paris, France
Ms DRANSART Estelle Estelle.Dransart@curie.fr
Cellular and Chemical Biology/Endocytic Trafficking and Intracellular Delivery
Institut Curie
Paris, FranceMr DRAY Nicolas nicolas.dray@pasteur.fr Department of Developmental and Stem Cell Biology Institut Pasteur, Paris, France Ms K'OUAS Guylène guylene.kouas@pasteur.fr Centre d'Enseignement, Institut Pasteur Paris, France Mr LEVAYER Romain romain.levayer@pasteur.fr Department of Developmental and Stem Cell Biology homéostasie des épithéliums Institut Pasteur Paris, France Mr LAMAZE Christophe Christophe.lamaze@curie.fr Cellular and Chemical Biology/Membrane Mechanics and Dynamics of Intracellular Signaling Institut Curie Paris, France Mr OBINO Dorian dorian.obino@pasteur.fr Department of Cell Biology and Infection Institut Pasteur Paris, France Mr SHAFAQ-ZADAH Massiullah massiullah.shafaq-zadah@curie.fr Cellular and Chemical Biology/Endocytic Trafficking and Intracellular Delivery Institut Curie Paris, France Mr THERY Manuel manuel.thery@cea.fr Physics of Cytoskeleton & Morphogenesis CEA, Hôpital Saint-Louis Paris – France Ms VIARIS Christine Christine.Viaris@curie.fr Cellular and Chemical Biology/Membrane Mechanics and Dynamics of Intracellular Signaling Institut Curie Paris, France
ADMINISTRATIVE CONTACTS
INSTITUT PASTEUR
Education Center
Gladys ELISABETH
Building 06 – Social building, Module 4
Tel: +33 (0)1 45 68 82 89
coursbmc@pasteur.fr
Registrar’s office
Sylvie MALOT, Céline CORBIN
Building 13, room St Joseph de Cluny
Tel: +33 (0)1 40 61 33 62
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