WP2: Building research capacity on telomere function in health and disease

Work package number

WP2

Work package title

Building research capacity on telomere function in health and disease

Objectives:

1. Exchange data, methodologies, and experimental models to broaden the scope of investigations

2. Share state-of-the-art infrastructure and equipment that is beyond the capacity of individual labs

Description of work

Task 2.1. Exchange data, methodologies, and experimental models to broaden the scope of investigations: The Azzalin lab has expertise in studying human telomere transcription and the role of lncRNA (TERRA) in supporting telomere stability through interactions with shelterin. The research team also specializes in molecularly characterizing the ALT mechanism and identifying vulnerabilities in human ALT cancer cells. The Garinis lab contributes expertise in translational research pipelines related to DNA damage, innate immune signalling, and telomere-driven metabolic abnormalities during aging. The team will provide TRIAD with mice that carry inborn deficiencies in transcription-coupled repair for the study of accelerated aging, telomere attrition, and premature onset of age-related diseases, e.g., chronic inflammation. The animal models will also serve as robust preclinical models for developing and testing drugs to combat degenerative diseases. In addition, the Garinis lab will provide expertise on the in vivo biotinylation tagging methodology to ensures the precise biotinylation tagging of physiologically expressed telomeric proteins for downstream methodologies (mass spectrometry, ChIP-Seq). The research team will also share its expertise on targeted nanocarriers loaded with modified mRNA- or protein-based drugs against chronic inflammation driven by telomere attrition. The Azzalin group will share their knowledge on the TALE system to suppress or upregulate TERRA transcription at will as well as on the Nanopore Long read sequencing approach to simultaneously identify and quantify human TERRA transcripts from all chromosome ends. The Krejčí laboratory specializes in biochemistry and biophysics, utilizing cutting-edge techniques to investigate telomere maintenance and HR-related processes. The Krejčí group will contribute its expertise in Stopped-flow technique for rapid kinetics of protein-DNA interactions at telomeric DNA, in BioLayer Interferometry for exploring protein-protein and protein-DNA interactions, shedding light on telomere-associated complex formation and stability, in Microscale Thermophoresis to determine binding affinities and thermodynamic parameters of macromolecular interactions related to telomere maintenance and in SwitchSENCE technology for studying telomeric proteins at the single-molecule level.

Task 2.2. Share state-of-the-art infrastructure and equipment that is beyond the capacity of individual labs:

The Garinis lab will offer TRIAD participants access to cutting-edge super-resolution microscopy that will allow the TRIAD research teams to observe and analyse biological structures at much higher resolutions as well as to 2-photon microscopy enabling visualization of living tissues at the cellular and subcellular level with high spatial resolution. The Azzalin group will provide access to state-of-the-art Flow Cytometry Facility that is equipped with multiple cell analysers and cell sorters. The Krejčí lab will grant access to Atomic Force Microscopy for visualizing biological samples with nanometric resolution, including analysis of nucleic acid-protein interactions as well as to Cryo-electron microscopy for visualizing telomeric structures and their interactions with other protein complexes at near-atomic or atomic resolution.