Periodic Reporting for period 1 - REVERSE (Circular RNAs to reverse pathological remodelling of the injured heart)
Reporting period: 2022-10-01 to 2025-03-31
Heart failure (HF) is a growing global health crisis, affecting over 60 million people worldwide. It is a leading cause of morbidity and mortality in industrialized nations. Current therapies for HF, while effective in some areas, fall short for specific subtypes of cardiac injury. For example, chemotherapy-induced cardiotoxicity and cardiac manifestations in COVID-19 patients represent emerging HF subforms for which no dedicated treatments exist. These conditions lead to irreversible damage to the heart, marked by cellular remodeling processes such as hypertrophy, inflammation, fibrosis, and apoptosis. The lack of effective therapies for these subtypes underscores the need for a deeper understanding of the underlying mechanisms driving HF.
The REVERSE project aims to tackle these challenges by focusing on circular RNAs (circRNAs), a novel and promising class of noncoding RNAs with critical roles in cardiac health and disease. CircRNAs have shown potential to reverse pathological changes in the heart, offering a groundbreaking avenue for therapeutic innovation. By developing circRNA-based therapies, REVERSE addresses unmet medical needs and improve the lives of millions suffering from HF.
The project pursues four key objectives:
1. Discovering Therapeutic Targets: Using advanced technologies, REVERSE will identify circRNAs involved in heart remodeling and damage caused by chemotherapy or COVID-19. By screening human cardiomyocytes with a CRISPR-Cas system designed specifically for circRNA silencing, researchers aim to uncover therapeutic targets that can reverse these processes.
2. Validating in Living Human Heart Tissue: A unique feature of REVERSE is its use of living human myocardial slices. This cutting-edge model preserves the heart's natural architecture and function, allowing researchers to study circRNAs in an environment closely resembling the human heart. This approach provides highly relevant insights into the therapeutic potential of circRNAs.
3. Uncovering Mechanisms of Action: Understanding how circRNAs work at a molecular level is crucial for developing effective therapies. REVERSE combines bioinformatics, proteomics, and advanced sequencing techniques to map interactions between circRNAs and their targets. This knowledge will pave the way for precision therapies tailored to specific HF subtypes.
4. Developing Targeted Delivery Systems: The project focuses on creating safe and effective delivery systems for circRNA-based therapies, including lipid nanoparticles and next-generation adeno-associated viruses (AAVs). These technologies ensure precise targeting to cardiac cells, minimizing side effects and enhancing therapeutic efficacy.
The REVERSE project aims to tackle these challenges by focusing on circular RNAs (circRNAs), a novel and promising class of noncoding RNAs with critical roles in cardiac health and disease. CircRNAs have shown potential to reverse pathological changes in the heart, offering a groundbreaking avenue for therapeutic innovation. By developing circRNA-based therapies, REVERSE addresses unmet medical needs and improve the lives of millions suffering from HF.
The project pursues four key objectives:
1. Discovering Therapeutic Targets: Using advanced technologies, REVERSE will identify circRNAs involved in heart remodeling and damage caused by chemotherapy or COVID-19. By screening human cardiomyocytes with a CRISPR-Cas system designed specifically for circRNA silencing, researchers aim to uncover therapeutic targets that can reverse these processes.
2. Validating in Living Human Heart Tissue: A unique feature of REVERSE is its use of living human myocardial slices. This cutting-edge model preserves the heart's natural architecture and function, allowing researchers to study circRNAs in an environment closely resembling the human heart. This approach provides highly relevant insights into the therapeutic potential of circRNAs.
3. Uncovering Mechanisms of Action: Understanding how circRNAs work at a molecular level is crucial for developing effective therapies. REVERSE combines bioinformatics, proteomics, and advanced sequencing techniques to map interactions between circRNAs and their targets. This knowledge will pave the way for precision therapies tailored to specific HF subtypes.
4. Developing Targeted Delivery Systems: The project focuses on creating safe and effective delivery systems for circRNA-based therapies, including lipid nanoparticles and next-generation adeno-associated viruses (AAVs). These technologies ensure precise targeting to cardiac cells, minimizing side effects and enhancing therapeutic efficacy.
1. Discovering Therapeutic Targets: A high-throughput screening system was established to identify circRNAs associated with chemotherapy-induced cardiotoxicity. Over 400 circRNAs were screened in iPSC-derived cardiomyocytes exposed to chemotherapeutic agents.
2. Validating in Living Human Heart Tissue: Studies using living human myocardial slices (LHMS) provided a physiologically relevant platform to evaluate chemotherapy effects. IC50 values for various agents were determined, and transcriptomic analyses are underway to validate candidate circRNAs. Improved handling of LHMS enabled further applicability to other disease contexts.
3. Mechanistic Characterization of circRNAs: Preliminary mechanistic insights are being explored to understand how identified circRNAs influence cardiac injury and recovery, laying the groundwork for targeted therapeutic development.
4. Development of Targeted Delivery Approaches: Progress was made in both viral and non-viral delivery systems. An AAV capsid library tailored to chemotherapy-affected cardiomyocytes and lipid nanoparticle (LNP) formulations demonstrated promising specificity and efficacy in preclinical models.
Overall, the project achieved significant technical and scientific milestones, establishing advanced screening systems, identifying therapeutic RNA targets, and developing innovative delivery technologies. These efforts pave the way for clinical applications in cardiac and other disease contexts.
2. Validating in Living Human Heart Tissue: Studies using living human myocardial slices (LHMS) provided a physiologically relevant platform to evaluate chemotherapy effects. IC50 values for various agents were determined, and transcriptomic analyses are underway to validate candidate circRNAs. Improved handling of LHMS enabled further applicability to other disease contexts.
3. Mechanistic Characterization of circRNAs: Preliminary mechanistic insights are being explored to understand how identified circRNAs influence cardiac injury and recovery, laying the groundwork for targeted therapeutic development.
4. Development of Targeted Delivery Approaches: Progress was made in both viral and non-viral delivery systems. An AAV capsid library tailored to chemotherapy-affected cardiomyocytes and lipid nanoparticle (LNP) formulations demonstrated promising specificity and efficacy in preclinical models.
Overall, the project achieved significant technical and scientific milestones, establishing advanced screening systems, identifying therapeutic RNA targets, and developing innovative delivery technologies. These efforts pave the way for clinical applications in cardiac and other disease contexts.
By achieving these objectives, REVERSE is expected to make significant contributions to global health:
• New Treatments for Unmet Needs: For HF subtypes caused by chemotherapy or COVID-19, REVERSE will provide the first targeted therapies, addressing critical gaps in current medical care.
• Reduced Animal Testing: The use of human myocardial slices aligns with ethical research practices, reducing reliance on animal models while enhancing the relevance of findings.
• Advancing Precision Medicine: By tailoring therapies to specific HF subtypes, REVERSE will demonstrate the potential of personalized medicine in combating complex diseases.
• New Treatments for Unmet Needs: For HF subtypes caused by chemotherapy or COVID-19, REVERSE will provide the first targeted therapies, addressing critical gaps in current medical care.
• Reduced Animal Testing: The use of human myocardial slices aligns with ethical research practices, reducing reliance on animal models while enhancing the relevance of findings.
• Advancing Precision Medicine: By tailoring therapies to specific HF subtypes, REVERSE will demonstrate the potential of personalized medicine in combating complex diseases.