Project description DEENESFRITPL Molecular mechanisms of human centromere stability maintenance Cell division depends on centromeres, which connect chromosomes to the spindle for separating sister chromatids in mitosis. Human centromeres consist of large arrays of repetitive DNA and are often sites of rearrangements in cancer. The molecular mechanisms that maintain repetitive DNA stability are poorly understood. The EU-funded Centromere Stability project will identify the human centromere maintenance network and investigate the mechanisms of repeats stability. The project will also study the consequences of centromere dysfunction, including changes in the size of the array, cell ploidy and proliferation dynamics. The proposed research will create a conceptual framework to explain the fragility of repetitive centromere DNA and the consequences for cell physiology and disease. Show the project objective Hide the project objective Objective Cell division relies on centromeres, which connect chromosomes to the spindle for separating sister chromatids in mitosis. Human centromeres are composed of large arrays of repetitive DNA, which are often sites of aberrant rearrangements in cancer. While centromere defects can cause chromosomal instability, the molecular mechanisms that maintain their repetitive DNA stable are poorly understood. During the fellowship, I aim to investigate how human centromere stability is maintained and the consequences of centromere dysfunction in driving cancer and aging. To circumvent impeding technical barriers due to incomplete centromere sequence annotation, I have ideated the use of Chromosome Orientation Fluorescence In Situ Hybridization at human centromeres (Cen-CO-FISH; Giunta, 2018). Using this innovative technique, I revealed that CENP-A and CCAN (constitutive centromere-associated network) proteins prevent centromere instability, and this functionality is compromised in cancer cell lines and in primary cells undergoing senescence (Giunta & Funabiki, 2017); my data show that CENP-A may play a new role during centromere replication, preventing DNA damage, repeats shortening, and subsequent aneuploidy. I will use the Auxin-Inducible Degron (AID) system and CRISPR-Cas genome editing with high-throughput imaging of Cen-CO-FISH to identify the human centromere maintenance network and investigate the mechanisms of repeats stability. I will also examine the consequences of centromeres dysfunction, including changes in the size of the array, cell ploidy and proliferation dynamics, using a variety of validated and novel methods, including Cen-qRT-PCR, qFISH and cytogenetic assays. Altogether, the proposed research will unveil a novel conceptual framework to explain the fragility of repetitive centromere DNA and its consequences on cell physiology and disease. This work will lay the foundation for my future independent research on centromere instability in age-associated cancers. Fields of science medical and health sciencesmedical biotechnologygenetic engineeringgene therapynatural sciencesbiological sciencesgeneticsDNAmedical and health sciencesclinical medicineoncologymedical and health sciencesbasic medicinephysiologynatural sciencesbiological sciencesgeneticschromosomes Keywords Centromere repetitive DNA genome stability recombination sister chromatid exchange chromosome-orientation fluorescence in situ hybridization (CO-FISH) senescence cancer aging aneuploidy Programme(s) H2020-EU.1.3. - EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions Main Programme H2020-EU.1.3.2. - Nurturing excellence by means of cross-border and cross-sector mobility Topic(s) MSCA-IF-2018 - Individual Fellowships Call for proposal H2020-MSCA-IF-2018 See other projects for this call Funding Scheme MSCA-IF - Marie Skłodowska-Curie Individual Fellowships (IF) Coordinator UNIVERSITA DEGLI STUDI DI ROMA LA SAPIENZA Net EU contribution € 183 473,28 Address Piazzale Aldo Moro 5 00185 Roma Italy See on map Region Centro (IT) Lazio Roma Activity type Higher or Secondary Education Establishments Links Contact the organisation Opens in new window Website Opens in new window Participation in EU R&I programmes Opens in new window HORIZON collaboration network Opens in new window Total cost € 183 473,28
