AnnualReport2020

DEVELOPMENT AND APPLICATION OF WELL-
DIFFERENTIATED CULTURE MODELS OF PRIMARY
AIRWAY EPITHELIUM FROM VARIOUS SOURCES
LE STUDIUM CONFERENCES
NOVEL HOST- AND MICROBIOTA-DIRECTED
STRATEGIES FOR TREATING RESPIRATORY
INFECTIONS
Life & Health Sciences 2020
42
Prof. Pieter Hiemstra
LE STUDIUM Research Professor
ARD 2020 BIOPHARMACEUTICALS Programme
From: Leiden University Medical Center - NL
In residence at: Centre for the Study of
Respiratory Pathologies (CEPR) -Tours
Nationality: Dutch
Dates:September 2018 to December 2018
September 2019 to October 2019
Pieter S. Hiemstra is Professor of Respiratory
Cell Biology and Immunology at the Department
of Pulmonology of the Leiden University Medical
Center (LUMC) in The Netherlands. He is
currently head of the Laboratory for Respiratory
Cell Biology and Immunology of the Department
of Pulmonology. His research is focused on basic
and translational research in chronic obstructive
pulmonary disease (COPD), asthma and lung
cancer. He has a specific interest in the role of
the epithelium of the airways and alveoli in these
diseases. His other activities include teaching and
curriculum development for Biomedical Sciences
and Medical students. Until October 2019, he
was head of Assembly 3 (Basic and translational
science) of the European Respiratory Society, and
he is currently the Section Editor of the European
Respiratory Journal. Pieter Hiemstra was elected
to a fellowship of the European Respiratory
Society in 2014.
Dr Mustapha Si-Tahar
Host Scientist
He is an INSERM research director who worked
for 10 years at the Pasteur Institute in Paris before
moving in 2012 to Tours to be the Head of Research
Center for Respiratory Diseases (french acronym:
“CEPR”). He also supervises one out of the three
research teams of this laboratory. M. Si-Tahar has
expertise in innate immunity and antimicrobial
responses in the lung mucosa as well as strong
competence with in vitro and in vivo models.Thus,
he demonstrated the role for specific patternrecognition
receptors and cell signalling pathways in
the immune response to major respiratory bacteria
and viruses, in the context of cystic fibrosis or
COPD. Besides, he contributes to several scientific
expert committees (including ANR, VLM) and is
the co-founder of Cynbiose Respiratory, a contract
research organization providing a set preclinical
services in the area of respiratory diseases.
This project is directly in link with the ARD2020 research programme
entitled «PRIMine – « Modèle préclinique de PRIMates non humains
(PNH) pour traiter les épisodes infectieux d’exacerbations de la Broncho-
Pneumopathie chronique obstructive (BPCO)». The specific aim of this
part of the project is to develop a state-of-the-art airway epithelial cell
(AEC) culture model from non-human primate tissue to study the effect
of cigarette smoke exposure and respiratory tract infections. This project
is supported by the ARD2020 Biopharmaceuticals Programme and this
report covers the work carried out in the second visit of my Professorship
in 2019: The set objectives were to:
1.Introduce the methods for culture of airway epithelial organoids
at CEPR with the aim to establish cultures of nasal epithelial cells
from nasal scrapings of non-human primates.
2.Further establish isolation and culture of primary airway human
epithelial cells at CEPR in collaboration with Thoracic Surgery (Dr
Antoine Legras) and Pathology clinical departments.
During the 2019 stay in Tours, to address aim 1 a nasal organoid
method was introduced in the laboratory that was developed in
the laboratory of the researcher in residence at LUMC. Using this
method, nasal epithelial cell cultures were established from nasal
scrapings derived from healthy volunteers and from non-human
primates. These 3D organoids cultures were next used to establish
2D air-liquid interface (ALI) cultures that are required to perform
exposures to airborne substances, such as respiratory viruses and
cigarette smoke. Organoid and ALI cultures were characterized using
immunofluorescence analysis of markers of epithelial differentiation.
To address aim 2, before and during the 2019 stay in Tours, methods
were provided and established at CEPR to begin setting up a biobank
of cultures of primary airway epithelial cells derived from resected
lung tissue. This proved to be successful, and a large number of vials
from several donors have now been stored in liquid nitrogen for the
establishment of the Tours biobank. Characterization for epithelial
cell markers and functional assays using influenza virus infections
were performed to confirm that such cultures established at the
CEPR behaved in a similar manner as those established at the home
institute of the researcher in residence (LUMC). The established
collaboration has resulted in a one-week training visit of one of the
researchers from CEPR (Virginie Vasseur) to the LUMC in Leiden in
June 2019. This visit was aimed at learning techniques required to
set up the above-mentioned biobank in of cultured human airway
epithelial cells in Tours.
Respiratory infections ranked
4 in the WHO 2016 list of “Top
10 causes of death worldwide”.
There is an urgent need for
improved treatment of respiratory
infections, in view of the absence
of affordable treatment, but
also because of the absence
of effective drugs for selected
respiratory pathogens and the
increase in microbial resistance to
conventional antibiotic treatment.
Recent insight into host defense
mechanisms against respiratory
infections and the respiratory
microbiota have opened up new
avenues for novel treatment
strategies.
The aim of this Le Studium Conference was to bring together experts from
a variety of European countries to discuss the latest developments in the
field, and to explore future opportunities for collaboration with the European
Horizon 2020 programme or related EU programmes.
This international conference was organised in the framework of the
BIOPHARMACEUTICALS ARD 2020 Programme.
This international conference was organised online in the framework of
the BIOPHARMACEUTICALS ARD 2020 Programme.
Life & Health Sciences 2020
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