Periodic Reporting for period 4 - TeloRNAging (The role of damage-induced non coding RNA in the control of DNA damage response activation at telomeres in aging)
Reporting period: 2024-04-01 to 2024-09-30
This project aimed to elucidate the role of telomeric non-coding RNA (tncRNA), which are transcribed following telomere dysfunction, in activating the DNA damage response (DDR). By leveraging our unique ability to selectively inhibit DDR at telomeres by suppressing tncRNA with antisense oligonucleotides (ASO), we identified the specific contribution of telomeric DDR activation to detrimental phenotypes associated with age-related disorders.
The results of this project are significant for society, as it provided evidence for a novel target to address age-related conditions, supporting the therapeutic potential of ASO - a class of drugs already in clinical use - to treat various diseases.
The project's objectives were as follows: to dissect the pathways controlling the transcription of dysfunctional telomeres, to investigate the molecular mechanisms through which tncRNA modulate DDR activation at telomeres, and to assess the impact of telomeric DDR inhibition on conditions of accelerated aging and age-related diseases.
The results of this project are significant for society, as it provided evidence for a novel target to address age-related conditions, supporting the therapeutic potential of ASO - a class of drugs already in clinical use - to treat various diseases.
The project's objectives were as follows: to dissect the pathways controlling the transcription of dysfunctional telomeres, to investigate the molecular mechanisms through which tncRNA modulate DDR activation at telomeres, and to assess the impact of telomeric DDR inhibition on conditions of accelerated aging and age-related diseases.
The results of this research explore the transcriptome of dysfunctional telomeres, focusing on telomeric non-coding RNAs (tncRNAs) and their role on DNA damage response (DDR) and aging. We analyzed RNA modifications that influence the interaction with DDR factors like 53BP1 and appear to affect DDR assembly and foci formation.
The study how tncRNAs and their interactors regulate DDR activation, we performed proteomic analysis of tncRNA-bound proteins in cells with induced telomere dysfunction. We identified RNA-binding the regions of 53BP1 that are crucial for phase separation and we are testing different mutants for their ability to form foci and allow DNA repair. Additionally, structural interactions between long and short RNAs were analyzed for their role in enhancing DDR activity.
Using a mouse model of aging, lacking telomerase activity and with critically short telomeres, (G3 Terc KO), we investigated the role of telomeric DDR in aging. These mice showed age-related diseases, including lung fibrosis and hematopoietic dysfunction. Treatment with telomeric antisense oligonucleotides (tASOs) reduced DDR activation, alleviated lung pathology, and restored tissue homeostasis. RNA sequencing confirmed tASOs normalized aging-associated transcriptional profiles, including pathways involved in inflammation and fibrosis. In hematopoietic organs, tASOs improved stem cell function, balancing proliferation and quiescence, and enhanced bone marrow reconstitution capacity. This research underscores the connection between telomere dysfunction, aging, and potential therapeutic interventions.
The study how tncRNAs and their interactors regulate DDR activation, we performed proteomic analysis of tncRNA-bound proteins in cells with induced telomere dysfunction. We identified RNA-binding the regions of 53BP1 that are crucial for phase separation and we are testing different mutants for their ability to form foci and allow DNA repair. Additionally, structural interactions between long and short RNAs were analyzed for their role in enhancing DDR activity.
Using a mouse model of aging, lacking telomerase activity and with critically short telomeres, (G3 Terc KO), we investigated the role of telomeric DDR in aging. These mice showed age-related diseases, including lung fibrosis and hematopoietic dysfunction. Treatment with telomeric antisense oligonucleotides (tASOs) reduced DDR activation, alleviated lung pathology, and restored tissue homeostasis. RNA sequencing confirmed tASOs normalized aging-associated transcriptional profiles, including pathways involved in inflammation and fibrosis. In hematopoietic organs, tASOs improved stem cell function, balancing proliferation and quiescence, and enhanced bone marrow reconstitution capacity. This research underscores the connection between telomere dysfunction, aging, and potential therapeutic interventions.
With these results, we developed a tool to separate telomere dysfunction from its effects - such as DDR activation, senescence, and aging - which represents a significant and groundbreaking discovery. We demonstrated that this tool can be used in vivo, in mouse models of age-related diseases, therefore it has the potential to be applied broadly to conditions linked to or driven by telomere dysfunction.