Intragranullar calcium regulation of mucin secretion and sorting.

Project Information

ICaRMuSs

Grant agreement ID: 894115

DOI

10.3030/894115

Project closed

EC signature date 9 March 2020

Start date 1 April 2020

End date 31 March 2022

Funded under

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Total cost

€ 172 932,48

EU contribution

€ 172 932,48

Coordinated by

FUNDACIO CENTRE DE REGULACIO GENOMICA
Spain

Periodic Reporting for period 1 - ICaRMuSs (Intragranullar calcium regulation of mucin secretion and sorting.)

Reporting period: 2020-04-01 to 2022-03-31

Summary of the context and overall objectives of the project

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).

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

Work was organized in 3 work packages (WP). WP1 comprised the generation and characterization of a CRISPR genetically-modified cell line to express a specific mucin tagged with a fluorescent protein. Several cell lines were generated an characterized. The data generated from these cell lines are going to be included in a publication for which the manuscript is being prepared and a second forthcoming publication. In addition these cell lines are going to be shared among scientists working in the field of mucus biology. These cell lines are also going to be used by me (fellow from hereon) as part of my future research as an independent investigator.
WP2 and 3 could not be performed as planned given an unsolvable problem with shared reagents. To circumvent this issue the fellow decided to characterize the role in mucin secretion of a protein for which no function was previously known. The fellow generated data that includes the interactome of the protein of interest, the effect of its ablation on mucin secretion and localization within the mucin-secreting cells. All these data are going to be part of the publication for which the manuscript is being prepared and a forthcoming publication.
For researcher training, the fellow assisted to 1 virtual course on microscopic analysis organized by the Crick Institute in the UK and another initiation course on proteomic analysis in the Center for Genomic Regulation. The fellow is also going to take part in an advanced course of proteomic analysis also to be taken in the Center for Genomic Regulation which had been postponed for limitations on people's gathering as a consequence of the COVID19 pandemic.
Although not included in the fellowship application, the fellow participated in the early response of the Catalan/Spanish government response to the COVID19 pandemic by taking part in the PCR screening of elderly people in residencies and hospitalized children (Ghose et al. F100Res. 2020).
For transfer of knowledge the fellow trained one under-graduate student. The training experience exceeded the expectations and the student became quickly independent and generated data that is going be included in the forthcoming publications. The student is going to be included as an author of the publication which is going to substantially contribute to her initiation steps in securing funding for her soon to begin PhD. The fellow trained several new members of the host laboratory in the CRISPR system and the methodology used to study mucin secretion and mucus formation. The fellow participated as an instructor in cell biology courses for PhD students in south america and is also participating in a PhD-postdoc mentoring program in the Center for Genomic Regulation in Catalonia/Spain which seeks to guide early PhD students in their scientific careers.
Results of this MSCA are reported in: (1) 2 oral presentations to the scientific staff of the Center of Genomic Regulation; (2) Oral presentation to researchers of the Institute of Medical Research in Argentina; (3) forthcoming papers on the regulation of mucin secretion; (4) forthcoming presentations in specific scientific congresses.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

This MSCA allowed the fellow to develop a deeper understanding and mastering of cutting-edge cell biology techniques such as CRISPR genetic engineering and super-resolution microscopy. It has also allowed the fellow to understand the importance and apply bio-informatic tools to analyze existing datasets to guide bench work much more efficiently. This learning and specialization are fundamental for the initial establishment of the fellow as an independent researcher in south america (Argentina) and the subsequent consolidation of the research group. As an independent researcher, the fellow will be responsible of student's initiation in science and cell biology research and subsequently in PhD careers. In addition, as an independent researcher in Argentina, the fellow will be required to share this knowledge and experiences with other researchers of the institute, country and region as well as to engage in events to disseminate scientific knowledge to the general public.
The fellow is now engaged in an initiative to promote research of understudied proteins, starting with the results and forthcoming publications of this action. This initiative has important researchers supporting it as evidenced by a recent open letter (Kustatscher et al. 2022). Future actions of the fellow are expected to increase the awareness of other researchers on the importance of dedicating efforts and resources to understand the functions of understudied proteins in all fields of cell biology. The impacts of this actions might have deep and highly ramifying societal implications as most current scientific publications are focused on a very narrow set of genes (@matthewhirschey).
The specific results and actions derived of this specific MSCA are also expected to rise scientist's and general public's awareness on the importance of understanding mucus formation in a time when air pollution has real impacts on human health.

Amazing group of people in the CRG that helpmed me during this MSCA

Enjoying science discussion in the amazing city of Barcelona