New insights in Progerin-induced modifications of chromatin landscapes

Genome structure, expression, and regulation are crucial in maintaining the physiological state of any cell. However, even a single variation in one of these processes can induce genomic instability, leading to different pathologies, including aging and cancer. Our work focuses on a particular model for of laminopathies disease, Hutchinson Gilford Progeria Syndrome, HGPS. HGPS is a genetic disorder in which patients show aging-related symptoms in the first years of their lives. HPGS is caused by a mutation in the Lamin A gene, LMNA, that causes the permanent anchoring of the mutated protein, named Progerin, to the nuclear membrane. This permanent bond causes abnormal tractions leading to a crumpled nuclear morphology that affects chromatin organization, inducing alteration in replication, transcription, and DNA repair. Indeed, one single point mutation in the LMNA gene results in massive damage to many cellular processes. Here, we exploit Structured Illumination microscopy, SIM, to investigate altered heterochromatin distribution in HGPS-model and compare the results with the control cell line. In particular, we explore how Progerin affects facultative heterochromatin organization in the proximity of the Nuclear Pore Complex, NUP, at the nuclear lamina level. To this aim, we perform multicolor SIM of DNA, Progerin or lamin (respectively HGPS or control), facultative heterochromatin (tagging histone H3K9me2) and NUP. Then, we combine SIM with Image Cross-Correlation Spectroscopy analysis, ICCS, to evaluate the differences in distances between NUP and H3K9me2. Our approach enables the visualization of Progerin-induced alterations of chromatin nanoscale organization at the single-cell level and will hopefully lead to a deeper understanding of the molecular mechanisms associated with HGPS development.