Descrizione del progetto
La regolazione del calcio nella secrezione della mucina
L’epitelio nelle vie aeree e nel tratto gastrointestinale è coperto da uno strato di mucosa prodotta da cellule mucipare caliciformi, che possono modificare le loro proprietà secretorie per adattarsi ai continui cambiamenti nell’epitelio. Le proteine più abbondanti nello strato di mucosa sono le mucine; difetti nella loro secrezione causano anomalie alle mucose, stress epiteliale e disfunzioni che possono condurre a infezioni microbiche, infiammazione cronica a e patologie più aggressive quali fibrosi cistica, asma e malattie polmonari ostruttive croniche. Il progetto ICaRMuSs, finanziato dall’UE, affronterà il modo in cui il calcio regola il trasporto, la selezione e la secrezione delle mucine, fornendo nuove informazioni sulla fisiologia del muco e determinando quali proteine di membrana sono importanti per la secrezione della mucina.
Obiettivo
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.
Campo scientifico
- medical and health sciencesclinical medicinepneumologyasthma
- natural scienceschemical sciencesinorganic chemistryalkaline earth metals
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteins
- medical and health sciencesbasic medicinepathology
- medical and health sciencesbasic medicinephysiology
Programma(i)
Argomento(i)
Meccanismo di finanziamento
MSCA-IF-EF-ST - Standard EFCoordinatore
08003 Barcelona
Spagna