1
MARIE SKLODOWSKA-CURIE ACTIONS
Co-funding of regional, national and international programmes (COFUND)
DOC2AMU THESIS PROJECT 2018 CALL FOR APPLICATIONS
Active response of Red Blood Cells to mechanical stress in splenic filtration
1. GENERAL INFORMATION
Call 2018-23
Topic Nano-health
Keywords Human red blood cells, splenic filtration, active volume regulation, microfluidics, live microscopy, computational modelling
2. THESIS DIRECTOR(S), RESEARCH UNITS AND DOCTORAL SCHOOLS
Thesis director Emmanuèle HELFER
Research Unit Centre Interdisciplinaire de Nanoscience de Marseille
Doctoral school ED 352 - Physique et Sciences de la Matière
Thesis co-director Catherine BADENS
Research Unit Génétique Médicale et Génomique Fonctionnelle
Doctoral school ED 062 - Sciences de la Vie et de la Santé
1
MARIE SKLODOWSKA-CURIE ACTIONS
Co-funding of regional, national and international programmes (COFUND)
DOC2AMU THESIS PROJECT 2018 CALL FOR APPLICATIONS
Active response of Red Blood Cells to mechanical stress in splenic filtration
(ActiveRed)
1. DESCRIPTION OF THE PHD THESIS PROJECT
1.1 OBJECTIVES OF THE PROJECT BASED ON THE CURRENT STATE OF THE ART
Blood consists in a highly concentrated suspension (45% in volume) composed mainly of red blood cells (RBCs,
99%) and few other blood cells (leukocytes and platelets, 1%). The efficient and sustainable circulation of RBCs
is an outstanding physical tour de force. During their 120-days lifespan, they continuously circulate through our
intricate microvascular network composed of slits, capillaries, bifurcations, etc. During such cycles they undergo
very strong deformations: for example, the RBC passes through blood vessels as small as 4 µm in diameter or
through submicron slits located in the spleen. The RBC being a biconcave disk around 2 µm in thickness and 8
µm in diameter, it cannot go through such constrictions if not highly deformable, and highly robust as well. This
is in part due to its complex double envelope that encloses the viscous haemoglobin solution. This double shell
is made of an incompressible fluid viscous lipid bilayer at the outside and a 2D elastic network of cross-linked
spectrin filaments at the inside, connected to the lipid bilayer by protein complexes. Additionally, a
mechanosensitive ion channel was recently discovered whose activation is triggered by a mechanical stress
applied on the RBC membrane [Coste 2012]. A second ion channel ion is then activated in cascade, leading to
water release out of the cell, as a way to control the RBC volume [Rapetti-Maus 2015]. A new hypothesis thus
arised that these ion channels could play an active role in RBC volume changes, in order to rapidly adjust the cell
deformability under a mechanical constraint.
The purpose of the ActiveRed project is to understand quantitatively the physical mechanisms of large
deformation and the molecular mechanisms of volume regulation in the specific case of the passage through
the splenic submicron slits. Recent studies indeed suggest that the spleen senses RBC deformability and
spheroidicity thus defining the size and shape of RBCs allowed to remain in the microcirculation [Pivkin 2016]. A
current hypothesis is that RBCs have to pass a ‘physical fitness test’ in the spleen, the submicron slits (Fig. 1A),
to be allowed to remain in the blood flow. Yet, no known physical mechanisms rationalize this hypothesis as
experiments are strongly lacking.
From a physics point of view, the process of splenic filtration raises basic physical questions: what is the link
between RBC mechanical parameters and passage/sequestration in splenic slits? What mechanical properties of
RBCs are most crucial to go through submicron splenic slits? Is the selection mechanism of RBCs by the spleen
based on mechanical criterions only? Or are indeed additional active phenomena, such as volume change,
needed to avoid RBC rupture under large deformation?
From a clinical point of view, there is a strong need for understanding the clearance process by the spleen. It
normally occurs as RBCs age and lose their deformability, to eliminate older RBCs and renew the RBC pool in the
blood stream. However, the spleen is also a major player in a number of diseases, whether infectious (malaria)
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or genetic (sickle cell disease (SCD), hereditary spherocytosis (HS), hereditary xerocytosis (HX)…). In all these
diseases, the RBC deformability is altered and RBCs get massively sequestered in the spleen. It thus causes severe
hemolytic anemia due to an accelerated splenic destruction of RBCs. Therefore, we expect that physics
experiments leading to understanding how an RBC passes the spleen fitness test and how the process is affected
in RBC genetic disorders will have a major impact in haematology.
Figure 1. A) An RBC (highlighted in red) squeezing through a splenic slit (≈0.5 x 2 x 5 µm3). B) SEM image of
the silicon master of a typical microfluidic device with a series of slits. C) Optical images of RBCs squeezing
through 0.8 x 1.9 x 5 µm3 biomimetic slits. The bottom one displays a tip while exiting the slit. Scale bar: 5 µm.
The ActiveRed project is based on the recent technological breakthrough we made in 2017, by fabricating a
microfluidic device with slits of physiological splenic dimensions (Fig. 1B) [Gambhire 2017]. This device allowed
us to observe human RBCs passing through these biomimetic slits and revealed new modes of deformation due
to high confinement (Fig. 1C, bottom). This is the first device that reproduces the dimensions of splenic slits (≈0.5
x 2 x 5 µm3) in comparison with previous biomimetic devices that could not reach such small dimensions [Rigat-
Brugarolas 2014, Deplaine 2011]. We will study the RBC behavior as they are submitted to controlled mechanical
stress (the slit dimensions and the flow pushing the RBCs). To investigate whether the RBC volume actively
changes in response to the stress, we will target the two ion channels which are thought to act together, the
mechanosensitive Piezo1 and the Ca2+-sensitive Gardos (Fig. 2A).
Figure 2. A) Schematics of Piezo 1 and Gardos interplay that controls ion fluxes: mechanical stress, e.g. RBC
stretching, activates Piezo1 which become permeable to cations, including calcium. The Ca2+ influx activates
Gardos leading to K+ and Cl- exit concomitantly with water. It results in a decrease in RBC volume, thus in an
increase in area-to-volume ratio, and presumably an increase in healthy RBC deformability. B) Activators
(arrows) and inhibitors of the two channels that will be used in the study.
Most studies on RBC ion channels are done on non human cells, mostly murine ones [Cahalan 2015], neglect
physiological flow and usually focus on one or the other channel. Here, modulation of Piezo1 and Gardos channel
activities will be studied in combination and not separately, in human RBCs, and in the physiological situation of
RBCs flowing though splenic slits. A recent work by the group of Kaestner studied their interplay using 3-µm wide
constrictions, they observed a response even at such small RBC deformation [Danielczok 2017]. We thus expect
a stronger response by using our biomimetic splenic slits.
The channels’ activity and interplay will be modulated via various combinations of known inhibitors and
activators (Fig. 2B). Healthy RBCs as wells as RBCs with disordered channels will be studied. Indeed, HX disease,
A B
ℓ= 1.9 µm 𝓌 = 0.8 µm
C
A B
Ca2+
Piezo 1 Gardos K
+
RBC
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due to various mutations in either Piezo1 or Gardos channels, is characterized by abnormal cation leak and cell
dehydration, leading to cell fragility and hemolytic anemia [Badens 2016]. We have access to a pool of patients
with some of these mutations. Their RBCs will be assayed in the biomimetic slits, untreated and treated with the
biochemical blocking/activating agents.
Our quantitative results will be combined with 3D computations (from international collaborator) that take into
account the RBC dynamics and the channels’ activity to derive the physical mechanisms responsible for RBC
active response to mechanical stress applied. Our findings will highlight which physical parameters can be used
as a new read out to follow disease evolution or treatment effect, and potentially lead to novel therapeutic
targets.
References:
Badens C and Guizouarn H. Advances in understanding the pathogenesis of the red cell volume disorders. Review.
Brit J Haematology 174:674-685 (2016)
Cahalan SM, Lukacs V, Ranade SS, Chien S, Bandell M, Patapoutian A. Piezo1 links mechanical forces to red blood
cell volume. eLife 4:e07370 (2015)
Coste B et al. Piezo1 and Piezo2 are essential components of distinct mechanically activated cation channels.
Science 330:55–60 (2010)
Danielczok et al. Red Blood Cell Passage of Small Capillaries Is Associated with Transient Ca2+-mediated
Adaptations. Front Physiol 8:979 (2017)
Deplaine et al. The sensing of poorly deformable red blood cells by the human spleen can be mimicked in vitro.
Blood 117:e88–e95 (2011)
Gambhire et al. High aspect ratio sub-micron channels using wet etching: Biomimetic spleen slits for red blood
cell studies. Small 13:1700967 (2017)
Pivkin IV et al. Biomechanics of red blood cells in human spleen and consequences for physiology and disease.
PNAS 113:7804–7809 (2016)
Rapetti-Maus et al. A mutation in the Gardos channel is associated with hereditary xerocytosis. Blood 126:1273-
1280 (2015)
Rigat-Brugarolas et al. A functional microengineered model of the human splenon-on-a-chip. Lab Chip 14:1715
(2014)
1.2 METHODOLOGY
The PhD program is pluridisciplinary and the student will learn the different required skills in the groups of the
two supervisors. The different tasks and milestones of the ActiveRed project are:
Task 0 (UMR_S910): Blood collection – Healthy and pathological blood sample collection will be obtained in the
context of regular clinical follow up at Hospital La Timone, and characterized.
Task 1 (UMR_S910): Characterization of Piezo1 and Gardos channel activity – Inhibitors and activators of Piezo1
and Gardos will be used to modulate RBC permeability (healthy and mutated). RBC ion content will be measured
under the various treatments to define the optimal combinations that will be assayed in the physics experiments.
Task 2 (CINaM): Relationship between RBC mechanical properties, RBC volume and channel activity as a function
of the applied mechanical stress – Prior to the microfluidic experiments, RBCs will be submitted to
meso/macroscopic deformations using atomic force microscopy (AFM) and optical tweezers. These techniques
allow applying forces in the pN-µN range. The resulting Ca2+ influx will be tracked using the commercial Fluo-4
calcium probe. Healthy and channel-deficient RBCs will be studied. From these measurements the
mechanosensitive characteristic response times will be extracted for healthy and patients’ RBCs.
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Task 3 (CINaM): Design and fabrication of new biomimetic devices – In the current device both the main wide
channel and the thin slits have the same height, thus RBCs are horizontally constrained before reaching the slits.
To get a geometry closer to that of the spleen, the device will be improved to have a main channel of larger
height (> 10 µm) with slits of 5 µm height.
Task 4 (CINaM + UMR_S910): Physical fitness test: relation between RBC deformation, transit time, flow rate,
channel activity, and slit size – Microfluidic experiments on individual RBCs flowing through biomimetic slits
(current and future devices) will be performed using ultrafast-videomicroscopy (> 1,500 fps) to observe RBC
deformation combined with standard fluorescence videomicroscopy (25 fps) to track calcium influx.
Experimental conditions (flow rate, slit dimensions) will be optimized so that the transit time is higher than the
response time derived from mesoscopic experiments and to be able to observe the calcium entry. The passage
of RBCs from healthy donors and from patients will be studied in absence and presence of the channel’s
modulators. The goal is to establish the laws of behaviour between the severity of the fitness test (given by slit
dimensions and flow rate) and RBC dynamics (shape deformation and velocity, ion fluxes).
Task 5 (CINaM + UMR_S910): PhD thesis writing and defense
Task 6 (Collaborator, Univ Notre Dame, USA): Modelling of the RBC active deformation under mechanical stress
– During the total duration of the project, we will communicate with our collaborator in the USA who will develop
a 3D model of the RBC that integrates the double envelope components, the channels, the resulting ion
transport, and the effect of the channel regulators. The coupling of experimental and numerical approaches will
lead to identification of the critical factors that cause RBC deformation, entrapment or damage, and investigate
how molecular mutations influence these critical factors. We expect to provide a complete physical
understanding of the dynamics of RBCs in spleen-like slits that will allow to predict the RBC behavior and the risk
of damage in case of altered mechanical properties. Moreover, we will conclude on the existence of an active
volume regulation in response to applied stress during spleen filtration.
1.3 WORK PLAN
1.4 SUPERVISORS AND RESEARCH GROUPS DESCRIPTION
Supervisor 1 – Centre Interdisciplinaire de Nanoscience de Marseille (AMU/CNRS, UMR7325, Marseille)
Emmanuèle HELFER joined CINaM in 2014, and now belongs to the newly created Physics and Engineering of
Living Systems (PIV) Department. The CINaM is a multidisciplinary structure, composed of approximately 180
persons, which hosts a nano/micro-fabrication platform that allows design and fabrication of complex
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36
Task0 Blood collection
Task 1 Characterization of channel's activity
Task 2 Characterization of RBC response
times (AFM, optical tweezers)
Task 3Design and fabrication of new
microfluidic devices
Task 4 Microfluidic experiments and analysis
Task 5 PhD writing and defense
Task 6
(collab)
Modelling of the RBC active
deformation under mechanical stress
DescriptionTasks
Months 1-12 Months 13-24 Months 14-36
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microfluidics devices. PIV department is equipped for cell biology, optical and near-field microscopies,
micropipette experiments, and microfluidics. E. Helfer works in the interdisciplinary field of physics of
macromolecular assembly since her PhD. She is an expert in in vitro biomimetic reconstitution of biological
processes, focusing on interactions between the actin cytoskeleton and cell membranes. In CINaM she goes on
developing a biomimetic approach of actin-based regulation of intracellular traffic. Since her arrival in Marseille,
she also worked in close collaboration with scientists from PIV Department to develop a microfluidic setup with
features mimicking closely the submicron-sized slits of the spleen that are dedicated to red blood cell (RBC)
filtration and selection. She is now interested in the molecular mechanisms occurring during RBC passage
through these biomimetic slits, i.e. the active regulation of the cell volume while it undergoes such a strong
deformation to squeeze in through the slit.
Supervisor 2 – Medical Genetics & Functional Genomics Unit (AMU/Inserm, UMR_S910, Marseille)
The UMR_S910 is presently staffed by approximately 100 people organized into 8 teams and is located at the
Campus Santé Timone (Medical School) of Marseille. Catherine BADENS is the head of the Molecular Genetics
Unit at the children hospital La Timone. She has a longstanding experience in medical biology and molecular
diagnosis of RBC genetic diseases, among which Sickle Cell Disease (SCD) and Hereditary Spherocytosis (HS).
These two diseases alter the mechanical properties of the RBCs leading to their sequestration in the spleen and
subsequent anemia. Recently, she has also characterized mutations in the Gardos ion channel, which result in
RBC dehydration. She now works on the combined action of this Ca2+-activated channel and the
mechanosensitive ion channel PIEZO1. She is designing various combinations of specific inhibitors of the channels
activity to modulate the hydration level of normal RBCs.
The project follows up a research program funded by A*Midex (RedPath, 2015-2017, coordinator: A. Viallat)
which allowed the fabrication of the biomimetic splenic-like slits.
2. 3I DIMENSIONS AND OTHER ASPECTS OF THE PROJECT
2.1 INTERDISCIPLINARY DIMENSION
The ActiveRed project involves two teams with the two supervisors attached to two Doctoral Schools of Aix-
Marseille University: ED352 (Physics and Matter Sciences) and ED62 (Life and Health Sciences). The PhD deals
with the fabrication of innovative microfluidic devices mimicking splenic slits, combination of high-speed video-
microscopy and fluorescence microscopy, image analysis, and manipulation of biological material. The PhD
student will participate in all the phases of the project, from the development of the devices, the microscopy
setup for dual observation, and the various combinations of biochemical treatments on RBCs to patients’
recruitment and sample collection.
E. Helfer has longstanding expertise in physics of the cytoskeleton and of cell membranes, and in using
biochemistry, cell biology and optical microscopy techniques to derive molecular interactions between actin
networks and cell membranes. She will lead the microfluidic, AFM and optical tweezers experiments and
subsequent analysis to extract RBC behavior changes under mechanical stress. C. Badens will provide molecularly
characterized healthy blood samples and samples affected with mutations in the Piezo and Gardos channels. She
will design various combinations of specific inhibitors and activators of the channels’ activity so as to modulate
the volume of normal RBCs in circulation. She will measure the cell ion content changes under the various
treatments to define the optimal combinations that will be assayed in the physics experiments. The experimental
results from RBCs under stress will be interpreted thanks to the input of the international collaborator (Z. Peng,
Univ Notre Dame, USA) who will inject the molecular elements of RBC in his computational model.
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The project is interdisciplinary by essence as it proposes a combination of know-how, expertise and knowledge
from different scientific fields and cultures: medical research, mechanics, nanotechnology and computation.
From our results we expect major advances in the global understanding of increased haemolysis and subsequent
anemia in RBC volume disorders induced by genetic mutations of Piezo 1 or Gardos channels.
2.2 INTERSECTORAL DIMENSION:
The non-academic partner is the Centre Hospitalier Universitaire La Timone, a hospital from Assistance Publique
des Hôpitaux de Marseille (APHM). La Timone is the most important hospital in PACA region and considered as
the 3rd european hospital from its activity, equipment and staff.
Dr Emmanuelle Bernit, member of the medical team of Department of internal medicine of APHM, focuses her
activity on hereditary anemias such as hemoglobinopathies and red cells membrane disorders. Around 200
patients with such blood genetic diseases are regularly monitored at the department she belongs to. Dr Bernit
will provide access to the cohort of patients, and specifically to those carrying mutations in the Piezo/Gardos
channels. The PhD student will participate in the recruitment process, i.e. meeting the patients and obtaining
their consent for providing their blood for the study. This clinical experience will be of benefit for her/him as it
will extend her/his understanding of the project from the fundamental research aspect to the clinical and ethical
one as well.
This ActiveRed project fits particularly with the Nano-Health axis of the SRI-S3 objectives. It aims at deciphering
the role of ion channels’ molecular interplay in the deformability and cell volume homeostasis of RBCs. Such
understanding is indeed of importance for chronic hemolytic anemia as it can open the way to development of
future treatments and diagnosis tools.
2.3 INTERNATIONAL DIMENSION:
We collaborate with Zhangli PENG, assistant professor at University Notre Dame (Indiana, USA), Within the PhD
period, the student will go visit his lab for one month to extend her/his knowledge to computing skills. Moreover,
she/he will attend once a year the annual APS DFD (Division of Fluid Dynamics) Meeting, the annual meetings of
either the British or the American Societies of Hematology, and at the national level the annual congress of the
Club du Globule Rouge et du Fer.
Dr Z. Peng has ten years’ experience in computational modeling of RBCs. Using advanced computational methods
he successfully predicted the bilayer-cytoskeletal interaction strength at the molecular level in RBCs and
highlighted the biomechanical mechanisms of hereditary spherocytosis and malaria transmission. He is currently
developing a multiscale model of RBCs with mechanosensitive channels flowing through splenic slits. In this
project, he will adapt his model to include the Ca2+-sensitive Gardos channel and compute RBC dynamics and
Ca2+/K+ transport at the cellular level. He will take into account the presence of agonists/antagonists or genetic
mutations of the two channels and explore their effects on RBC volume control. He will compare the predicted
cell deformation and ion transport with experiments for validations. The synergistic effects from our
complementary skills in microfluidic experiments on RBCs and multiscale simulations will enable the realization
of the ActiveRed project.
3. RECENT PUBLICATIONS
Relevant publications and patents of Supervisor 1: Dr Emmanuèle HELFER
1. Ghambire P, Atwell S, Iss C, Badens C, Helfer E, Viallat A, Charrier A. High aspect ratio sub-micron channels
using wet etching: Biomimetic spleen slits for red blood cell studies. Small 13:1700967 (2017)
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2. Rommelaere S, Millet V, Rihet P, Atwell S, Helfer E, Chasson L, Beaumont C, Chimini G, do Rosário Sambo M,
Viallat A, Penha-Gonçalves C, Galland F, Naquet P. Serum pantetheinase/Vanin levels regulate erythrocyte
homeostasis and severity of malaria. The Am J Pathol 185:3039-3052 (2015)
3. Helfer E, Harbour ME, Henriot V, Lakisic G, Sousa-Blin C, Volceanov L, Seaman M, Gautreau A. Endosomal
recruitment of the WASH complex: active sequences and mutations impairing interaction with the retromer. Biol
Cell 105:1-17 (2013)
4. Derivery E, Helfer E, Henriot V, Gautreau A. Actin polymerization controls the organization of WASH domains
at the surface of endosomes. PLoS One 7:e39774 (2012)
5. Delatour V, Helfer E, Didry D, Lê KHD, Gaucher JF, Carlier MF, Romet-Lemonne G. Arp2/3 controls the motile
behavior of N-WASP-functionalized GUVs and modulates N-WASP surface distribution by mediating transient
links with actin filaments. Biophys J 94:4890-4905 (2008)
Relevant publications and patents of Supervisor 2: Prof Catherine BADENS
1. Rapetti-Mauss R, Picard V, Guitton C, Ghazal K, Proulle V, Badens C, Soriani O, Garçon L, Guizouarn H. Red
blood cell Gardos channel (KCNN4): the essential determinant of erythrocyte dehydration in hereditary
xerocytosis. Haematologica 102:e415-e418 (2017)
2. Rapetti-Mauss R, Soriani O, Vinti H, Badens C, Guizouarn H. Senicapoc: a potent candidate for the treatment
of a subset of hereditary xerocytosis caused by mutations in the Gardos channel. Haematologica 101:e431-e435
(2016)
3. Badens C and Guizouarn H. Advances in understanding the pathogenesis of the red cell volume disorders.
Review. Brit J Haematology 174:674-685 (2016)
4. Rapetti-Mauss R, Lacoste C, Picard V, Guitton C, Lombard E, Loosveld M, Nivaggioni V, Dasilva N, Salgado D,
Desvignes JP, Béroud C, Viout P, Bernard M, Soriani O, Vinti H, Lacroze V, Feneant-Thibault M, Thuret I, Guizouarn
H, Badens C. A mutation in the Gardos channel is associated with hereditary xerocytosis. Blood 126:1273-80
(2015)
5. Patent. Badens C, Guizouarn H, Thuret I. Diagnostic and treatment of Hereditary Xerocytosis. Property: APHM,
AMU. Patent # EP 15305921.7
4. EXPECTED PROFILE OF THE CANDIDATE
The applicant must have a master degree in experimental physics or biology, with a strong interest towards
biophysics. A background in optical microscopy and/or basic knowledge in biology, though not required, will be
welcome.
The PhD student is expected to be sociable and eager to broaden her/his interdisciplinary knowledge. She/he
will have to interact with physicists, biologists, biophysicists and physicians. Skills in image analysis and
programming (Image J, Matlab, Python) will be acquired during the PhD period.
5. SUPERVISORS’ PROFILES
Supervisor 1:
Emmanuèle HELFER was born in Nancy (France) in 1973. She did her studies at the University of Strasbourg
where she obtained her PhD in 1999 in Biophysics, on the viscoelastic properties of actin-coated vesicles, under
the supervision of D. Chatenay and L. Bourdieu. In 2001 she obtained a CNRS position in the 'Cytoskeleton and
Cell Motility' group, in the Laboratoire d'Enzymologie et Biochimie Structurales (LEBS, Gif-sur-Yvette, France)
where she worked on biomimetic reconstitution of actin-based motile processes. In 2010 she moved to the
'Cytoskeleton and Cell Morphogenesis' group (LEBS) where she studied the role of actin structures on
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intracellular endosomal membranes by combining cell biology and biochemical approaches. End of 2014, she
moved to Marseille (France) to join CINaM, where a new department, 'Physics and Engineering of Living Systems'
(PIV), was being created and she actively participated in its establishment (official start in January 2018). In PIV
department she develops her biomimetic approach to decipher further the role of endosomal actin in
intracellular traffic. On the other hand, she started collaborations with the other scientists of the group on red
blood cell dynamics and mechanics, and on cell adhesion. Her involvement in the RBC project has been increasing
since then and she is now taking the leadership in aspects related to molecular regulation.
E. Helfer has published 20 papers in peer-reviewed journals and 2 book chapters, in the domain of actin-
membrane interactions and actin biochemical regulation. She gave 3 invited talks, 4 oral presentations at
international conferences, and 2 at national ones. She has been funded an ANRJCJC (COORDACTIN, 2010-2012),
a local grant from PACA Région (BioMimWASH, 2017-2018) and one from A*Midex in which she is leading the
CINaM participation (MecaLam, 2018-2020; coordinator: C. Badens). She has been requested to participate in
committees for faculty selection (Paris-Diderot Univ, 2009-2011) and laboratory evaluation (LJP Hcéres
committee, 2018), and reviewed 2 PhDs in 2017 (ENS Chimie, Paris; Institut Jacques Monod, Paris). At the
national level, she is member of the National committee of the CNRS (2016-2021), in Section 11 (“Supra and
macromolecular systems and materials: design, properties, functions”) where she is one of the experts in the
field of physics-biology interface.
She obtained in 2014 her Accreditation to Supervise Research in Biochemistry and Cell Biology (Paris-Sud
University), on The interactions between the actin cytoskeleton and membranes, before she moved to Marseille.
She has been co-supervising 1 PhD thesis (2003-2007, Vincent Delatour) leading to 2 publications [New J Phys,
2008; Biophys J, 2008] and 1 review [Biophys Rev & Lett, 2009]. V. Delatour obtained a position in 2008 in
Laboratoire National de Métrologie et d'Essais (LNE, Paris). Since October 2017, she is co-supervising a PhD
student with Dr A. Viallat (PIV) on the vaso-occlusion mechanism in the pathological context of Sickle Cell Disease
where RBCs become less deformable and adhesive, and thus form aggregates. She will supervise a PhD student
from October 2018 on a new project on cell mechanics in premature aging diseases funded by A*Midex. She (co)-
supervised 12 master students since 2001, among them 3 were supervised and 3 were co-supervised since her
arrival in CINaM.
Supervisor 2:
Catherine BADENS is Pharm D, DES Biologie Médicale (1991), PhD(1996) and did her studies at Aix Marseille
University.
She is the head of the Molecular Genetics Unit at the children hospital La Timone and head of the Laboratory of
Biochemistry in La Conception, both hospitals located in Marseille city center. She has a longstanding experience
in medical biology, molecular diagnostic in rare diseases, patients registry and translational research projects.
Since October 2011, she is Professor of Biological Sciences, at the Faculty of Pharmaceutical Sciences, Aix
Marseille University, France. She is the scientific coordinator of the French Registry for Thalassemia and a
member of the board of the French Centre of reference for Thalassemia (http://www.chu-lyon.fr/web/2652 ).
She has authored or co-authored 104 papers in peer-reviewed journal and is co-inventor of 3 patents (pending).
She has supervised 3 PhD thesis, all 3 are now working at APHM (1 PH, 1 engineer) or AMU/APHM (1 MCU/PH).
She has recently been funded a grant from A*Midex as the coordinator: MecaLam, 2018-2020.
DD EE PP AA RR TT EE MM EE NN TT DD EE MM EE DD EE CC II NN EE II NN TT EE RR NN EE
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Docteur Emmanuelle BERNIT
Praticien Hospital ier
RPPS : 10003428785
Docteur Nicoléta ENE
Praticien Hospital ier
RPPS : 10004415906
Docteur J. François LAMARCHI
Praticien Hospital ier
RPPS : 10003431458
Docteur Christine NICOLINO
Praticien Hospital ier
RPPS : 10003346698
Docteur Véronique VEIT
Praticien Hospital ier
RPPS : 10003376133
Docteur Laure SWIADER
Praticien Hospital ier
RPPS : 10003370011
Docteur Estelle JEAN
Praticien Hospital ier
RPPS : 10100410215
Docteur Mikaël EBBO
Maître de Conférence des Universités
PH -RPPS : 10100287019
Docteur Pauline BELENOTTI
Assistant Chef de Cl inique
RPPS : 10100613701
Docteur Jacques POZZO di BORGO
Assistant Chef de Cl inique
RPPS : 10100686459
Docteur Aurélie GRADOS
Assistante Chef de Cl inique
RPPS :10100842276
Docteur M. Pierre DI COSTANZO
Vacataire
RPPS : 10003358172
Accueil Consultations Externes
Tél : 04 91 38 60 33
NUMADIC : Numéro d’Aide au Diagnostic
Du lundi au vendredi de 9 h à 18h Tél : 04 91 38 79 00
Marseille, le 06/01/2018
As a member of the medical team of Department of internal medicine of the
“Assistance Publique des Hôpitaux de Marseille”, I am supporting a biological physics
project entitled:
“Active response of red blood cells to mechanical stress in splenic filtration”
The physics student will conduct this project under the co-supervision of a physicist, Dr
Emmanuele HELFER and a biologist, Prof. Catherine BADENS. I am deeply convinced
that in the context of his/her doctoral training, an experience in clinical laboratory and a
reference center for rare diseases would be very beneficial for his/her project by enabling
a global understanding of the topic.
Our department participates in several research projects including clinical trials with
industrial or academic sponsors. My main activity focuses on hereditary anemias such as
hemoglobinopathies and red cells membrane disorders. A cohort of a bout 200 patients
are monitored regularly at our hospital. Anemia observed in such diseases is partly due to
splenic sequestration of red cells whose deformability is altered for further destruction.
An unmet need does exist for a better understanding of red cells deformability in chronic
hemolytic anemia and of the implication of red cells volume homeostasis via ion
channels. Advances in this field will certainly lead to the identification of new targets for
future innovative treatments and to more powerful diagnosis tools. This could be achieved
by interdisciplinary projects such as the one we are supporting here. The project aims in
deciphering the mechanisms at play during the passage of red cells through splenic
filtering slits, and more specifically the ones involving ion channels regulating the cell
volume.
The success of this project depends in part on the access to the largest number of blood
samples. These samples are collected in our department on clinical research participants
after their appropriate consent.
I deeply think that the student should participate in this essential step of the research:
meeting the medical team and discussing about his/her project and the expected outputs
will surely motivate him/her. Furthermore, it would raise the student’s awareness about
ethical, legal and clinical aspects of translational research. The student time dedicated to
this aspect of the project with the medical staff, will be approximately one day per month.
Thank you in advance for your attention to this project,
Sincerely,
Dr Emmanuelle Bernit