Description du projet
La régulation de la sécrétion de mucines par le calcium
L’épithélium des voies respiratoires et de l’appareil digestif est recouvert d’une couche de mucus produite par les cellules caliciformes, capables de modifier leurs propriétés sécrétoires pour s’adapter aux changements incessants de l’épithélium. Les protéines les plus abondantes dans cette couche de mucus sont les mucines. Les défauts dans la sécrétion de mucines provoquent des anomalies dans le mucus, ainsi que le dysfonctionnement et le stress épithélial pouvant entraîner des infections microbiennes, une inflammation chronique et des pathologies plus agressives telles que la mucoviscidose, l’asthme et la bronchopneumopathie chronique obstructive. Le projet ICaRMuSs, financé par l’UE, abordera la manière dont le calcium régule le trafic, le tri et la sécrétion de mucines, en apportant de nouvelles informations sur la physiologie du mucus et en déterminant quelles sont les protéines membranaires importantes pour la sécrétion des mucines.
Objectif
The epithelium in the airways and gastrointestinal track are covered by a mucus layer which protects the epithelium from mechanical and microbial insults. The most abundant proteins in the mucus layer are mucins which are synthesized and secreted by goblet cells. Mucus function is largely dictated by mucin composition and goblet cells can modify their secretory properties to adapt to the continuous changes the epithelium is encountered with. Mucins are heavily glysosylated and packed into secretory vesicles in the Golgi apparatus. A calcium signal triggers the fusion of the secretory vesicles with the plasma membrane releasing mucins to the extracellular space. The role of intragranular calcium remains a mayor gap in our understanding of mucin physiology and how goblet cells precisely control mucus composition remains only a matter of speculation. These are highly relevant questions as defects in mucin secretion cause mucus abnormalities, epithelial stress and dysfunction that can lead to microbial deleterious infections, chronic inflammation and more aggressive pathologies such as cystic fibrosis, asthma, and chronic obstructive pulmonary disease.
In this project I will address how calcium regulates the trafficking, sorting and secretion of mucins, filling current gaps in our understanding of mucus physiology. I will genetically engineer mucin-secreting cells with the CRISPR/Cas9 tools to add GFP and RFP to MUC5AC and MUC2 loci and describe granular composition by super resolution microscopy and FACS. I will use a first-in its kind probe to measure intragranular calcium concentration and finally I will purify secretory granules and sequence them with mass spectrometry to determine which membrane proteins are important for mucin secretion. This project will allow me to apply my experience in highly relevant questions in cell biology and it will also enhance my creative and innovative potential through advanced training and international mobility.
Champ scientifique
- medical and health sciencesclinical medicinepneumologyasthma
- natural scienceschemical sciencesinorganic chemistryalkaline earth metals
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteins
- medical and health sciencesbasic medicinepathology
- medical and health sciencesbasic medicinephysiology
Programme(s)
Régime de financement
MSCA-IF-EF-ST - Standard EFCoordinateur
08003 Barcelona
Espagne