DC9
Compartmentalized ciliary proteome analysis (WP3)
Supervisor: Dr Karsten Boldt
Host Institute: University of Tübingen, Germany
Secondments: Radboud University Medical Center Nijmegen, The Netherlands; University of Copenhagen, Denmark
Doctoral Program: University of Tübingen
I am Marco Tapia from Peru, with a B.Sc. in Biology and an Erasmus Mundus M.Sc. in Sustainable Drug Discovery, awarded jointly by Ghent University, the Medical University of Gdańsk, the University of Lille, and the University of Groningen. During my master's thesis. I investigated pharmacological strategies to enhance lung regeneration in chronic obstructive pulmonary disease (COPD). Using ex vivo mouse lung slices and in vitro organoid models of emphysema and cigarette smoke exposure. To explore underlying mechanisms, I conducted transcriptomic RNA-seq analysis. As part of the Cilia-AI network, I am excited to expand my expertise in omics and spatial proteomics to study the compartmentalized proteome of cilia and advance our understanding of their role in human disease.
Compartmentalized ciliary proteome analysis
Cilia are highly compartmentalized structures. These compartments, namely basal body, transition zone, axoneme and tip, are characterized by a distinct protein composition that is dynamically altered, depending on the ciliary state. To date, a comprehensive definition of the compartmentalized proteome and the changes induced by disease is lacking. By employing and adapting state of the art proximity-labelling technologies (TurboID or APEX), this project aims at defining the protein composition of the ciliary compartments. To this end, endogenously tagged cell lines with proximity-labelling tags on specific marker proteins will be developed to dissect the whole ciliary proteome, the ciliary tip proteome as well as the basal body proteome as well as the transition zone. The protein composition of these compartments as well as its alterations upon knock-out of ciliopathy associated proteins or knock-in of disease-associated mutant (i.e. LCA5, CEP290, RPGRIP1L) will be quantitatively analysed by mass spectrometry. Data analysis and integration with pre-existing data like the ciliary landscape will lead to a detailed understanding of the ciliary mechanisms, leading to disease.