Resolving the mechanism of ciliary tip factors in primary and motile cilia assembly and function

Informations projet

ResolvCiliaTip

N° de convention de subvention: 896644

DOI

10.3030/896644

Projet clôturé

Date de signature de la CE 22 Avril 2020

Date de début 1 Juin 2020

Date de fin 31 Mai 2022

Financé au titre de

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Coût total

€ 145 355,52

Contribution de l’UE

€ 145 355,52

Coordonné par

KOC UNIVERSITY
Türkiye

Periodic Reporting for period 1 - ResolvCiliaTip (Resolving the mechanism of ciliary tip factors in primary and motile cilia assembly and function)

Période du rapport: 2020-06-01 au 2022-05-31

Résumé du contexte et des objectifs généraux du projet

The Action “Resolving the mechanism of ciliary tip factors in primary and motile cilia assembly and function” aims to find out how some of our cells build non-motile cilium, a tiny single projection on their surfaces to catch the signals from the environment like an antenna (photoreceptors in eyes). Another goal is to find out how other cells build motile cilium, which can propel that cell (sperm cell) or how do they build multiple motile cilia that move fluid around them (and in that way clear mucus from the lungs or move the egg to the uterus). This is medically important because the absence of cilia or the errors in their construction and content can cause a wide range of human diseases, collectively called ciliopathies, and they can affect up to 1 in 2000 people. These diseases can affect human development and multiple organs and systems in our bodies. They are caused by mutations in specific genes. Although many genetic causes of ciliopathies are known, a not-so-small proportion of ciliopathy cases remains without a genetic diagnosis. Sometimes even if the mutations in genes that cause ciliopathies are known, it is not clear how the protein products of those genes build cilia or control their functions.
The basic structure of both non-motile and motile cilia is very similar, as they contain the main central cytoskeletal scaffold and membrane around it. How this cytoskeletal scaffold is built and maintained is still not completely understood, and yet, the defects in its assembly and organization are the major causes of ciliopathies. Objectives of this Marie Skłodowska Curie Action (MSCA) have been to resolve: (1) how the proteins that cause ciliopathies build the ciliary scaffold and how this affects the function of both non-motile and motile cilia; (2) how they control the overall stability of that ciliary scaffold and the very tip of cilia; and (3) to identify the protein content of non-motile and motile ciliary scaffold and find the potential new causes shared between primary and motile ciliopathies. Another important goal of the MSCA Individual Fellowship is to foster the development of this individual researcher. The fellow and the researchers in the host laboratory resolved the localization and function of ciliopathy-related protein in detail in both non-motile and motile cilia, as well as synergistic action with other important proteins for cilia formation and function.

Travail effectué depuis le début du projet jusqu’à la fin de la période considérée dans le rapport et principaux résultats atteints jusqu’à présent

The work done by the fellow together with the host laboratory detailed the localization of the ciliopathy-related protein in non-motile cilia and described the role in cilia assembly, maintenance, ciliary trafficking and signalling, and distribution of other Joubert syndrome-associated proteins. We discovered that this protein directly stabilizes the ciliary scaffold and identified its new co-operations with other proteins in cilia. Excitingly, we discovered that this protein has a similar stabilization role in cell division, which could also contribute to the development of ciliopathies (such as kidney cysts). Moreover, our preliminary results discovered the specific localization and function of this protein in motile cilia. Therefore, this project identified a novel regulator of both non-motile and motile cilia, thus shedding a light on the novel connection between primary and motile ciliopathies. The fellow received extensive training in state-of-the-art techniques related to the field, as well as academic training in project management, grant application (during the grant, the fellow has secured three governmental grants which totals €100,000), research presentation, supervision of the graduate students, and lecture preparation.
The results of this MSCA are available with open access on bioRxiv (doi:10.1101/2021.07.20.453048; doi:10.1101/2022.04.10.487777; doi:10.1101/2022.04.22.489036) published in one peer-reviewed journal (doi:10.1111/febs.16367) while the other article is currently under review. One published article in a peer-reviewed journal with open access was an addition to this action (doi.org/10.1371/journal.pbio.3001708). The data and datasets collected during this MSCA will inform more publications (expected three publications) in the coming year, in addition to the ones produced and published during the period of the fellowship. The results were presented at two virtual conferences, one in person and one in the following two months.

Progrès au-delà de l’état des connaissances et impact potentiel prévu (y compris l’impact socio-économique et les conséquences sociétales plus larges du projet jusqu’à présent)

This action expanded scientific knowledge about cilia formation and regulation. State-of-the-art technology was applied to the cilia field for the first time, such as miniTurboID protein proximity labelling and TIRF microscopy. This shed a light on the novel connection between non-motile and motile ciliopathies. The result of this MSCA could influence the development of new diagnostic tools and therapeutic approaches for these diseases. The potential impact that we are anticipating (and hoping for) is that this research will improve the quality of life of ciliopathy patients.

The microscopically small and interesting world of my research

Midway during the lab effort to purify tubulin from bovine brain at Kurban Bayram Saturday

The EMBO Microtubules: from atoms to complex systems, my first experience of an online conference.

When the restrictions eased up I was happy to attend the conference EMBO Centrosomes meeting.

Me and my weekly microscope routine taken for the lab Instagram account.

This is what isolated motile cilia looks under a microscope, with colourful fluorescent proteins

Tubulin successfully purified and aliquoted in our cold room. Big smiles at the end of the long day.