ICaRMuSs · Intragranullar calcium regulation of mucin secretion and sorting.
Horizon 2020 — Marie Skłodowska-Curie Actions
- Duration
- 2020-04-01 → 2022-03-31
- EU contribution
- €172,932
- Participants
- 1
- Scheme
- MSCA-IF-EF-ST
Lines connect the coordinator with its partners.
Results in brief
Intragranullar calcium regulation of mucin secretion and sorting.
The action "Intragranullar calcium regulation of mucin secretion and sorting" aims to elucidate how cells in the respiratory and digestive systems generate the mucus layer that protects those tissues. The mucus is a thick viscous-elastic layer that sits on top of mucous epithelium. In the lungs it maintains the epithelium free from contaminant particles and microorganisms while in the intestine it maintains a micro-environment for symbiotic bacteria to live in. In both tissues the mucus layer has a protective role and its alteration may promote infections, airways obstruction or poor nutrient absorption.The most abundant proteins of the mucus layer are called mucins and these proteins confer most of its biophysical properties. Although some aspects of mucin secretion are known many questions remain unanswered. One very important aspect we still don't fully understand is how the amount of mucin released is regulated and this is the main issue addressed in this action. In the last few decades rural populations are migrating to cities in search for better economic opportunities and quality of living. This trend is accelerating at a global scale. At the same time, air, water and soil pollution is increasing at an alarming rate and negatively impacting human health. This is causing preventable premature deaths every year (https://www.theguardian.com/environment/2022/may/17/pollution-responsible-one-in-six-deaths-across-planet). The mucus layer is the actual interface that mediates substances exchange between the human body and the environment it inhabits. Understanding how this layer is generated and maintained is not only important for basic scientific knowledge itself but also of utmost importance when trying to mitigate the negative effects pollution has on humans. The objectives of this Marie Skłodowska Curie Action (MSCA) have been to (1) describe the cellular and molecular mechanisms that regulate mucin secretion; (2) motivate awareness of the importance of this topic on other scientists and students; and (3) to train students in the methodology and literature relevant to this topic and to motivate them to engage in specific research to better understand mucus layer generation and maintenance. Additionally, a parallel goal of this action is to foment the personal and scientific development of the fellow (me).
Data: CORDIS, © European Union
Project objective
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.
Original text from CORDIS.
Participants
- FUNDACIO CENTRE DE REGULACIO GENOMICA · BarcelonaCoordinatorSpain
Links
Data: CORDIS, © European Union