Periodic Reporting for period 1 - MERAS (Mechanoregulation of alternative splicing - a multi-omics and single cell approach to improved cardiac function)
Periodo di rendicontazione: 2023-01-01 al 2025-06-30
Heart failure represents a growing global health burden, significantly impacting patient quality of life and healthcare systems. A prominent form, heart failure with preserved ejection fraction (HFpEF), remains poorly understood, lacking effective therapeutic options. Central to heart function is the sarcomere, the basic unit responsible for heart contraction, where the giant protein titin plays a critical role. Titin not only influences muscle elasticity but also participates in sensing mechanical stress, signaling to the nucleus, and subsequently affecting gene expression through a process known as mechanotransduction.
The MERAS project (Mechanoregulation of Alternative Splicing) investigates how mechanical signals from the heart muscle affect the alternative splicing of genes—an essential regulatory mechanism that allows a single gene to generate multiple protein variants. Misregulation of alternative splicing significantly contributes to cardiac diseases. MERAS utilizes state-of-the-art multi-omics approaches, combining advanced proteomics, transcriptomics, and single-cell sequencing to unravel the mechanistic links between mechanical stress and splicing regulation.
The overall goal of MERAS is to map how mechanical forces within cardiac muscle cells regulate alternative splicing, and identify how disruptions in these pathways contribute to heart disease. The project employs engineered heart tissues (EHTs) derived from human induced pluripotent stem cells (hiPSCs), allowing precise control of mechanical and environmental conditions.
By integrating machine learning techniques for data analysis, MERAS researchers can identify patterns and interactions within extensive datasets, significantly enhancing the understanding of cardiac mechanotransduction. Such insights are crucial for developing targeted therapeutic strategies.
Ultimately, MERAS aims to translate foundational biological discoveries into clinical innovations. The identification of new molecular targets and mechanisms promises to guide future drug development, addressing a critical need in cardiology. By bridging basic science and clinical application, MERAS aims to improve our ability to manage and treat heart failure and enhance patient outcomes.
The MERAS project (Mechanoregulation of Alternative Splicing) investigates how mechanical signals from the heart muscle affect the alternative splicing of genes—an essential regulatory mechanism that allows a single gene to generate multiple protein variants. Misregulation of alternative splicing significantly contributes to cardiac diseases. MERAS utilizes state-of-the-art multi-omics approaches, combining advanced proteomics, transcriptomics, and single-cell sequencing to unravel the mechanistic links between mechanical stress and splicing regulation.
The overall goal of MERAS is to map how mechanical forces within cardiac muscle cells regulate alternative splicing, and identify how disruptions in these pathways contribute to heart disease. The project employs engineered heart tissues (EHTs) derived from human induced pluripotent stem cells (hiPSCs), allowing precise control of mechanical and environmental conditions.
By integrating machine learning techniques for data analysis, MERAS researchers can identify patterns and interactions within extensive datasets, significantly enhancing the understanding of cardiac mechanotransduction. Such insights are crucial for developing targeted therapeutic strategies.
Ultimately, MERAS aims to translate foundational biological discoveries into clinical innovations. The identification of new molecular targets and mechanisms promises to guide future drug development, addressing a critical need in cardiology. By bridging basic science and clinical application, MERAS aims to improve our ability to manage and treat heart failure and enhance patient outcomes.
The MERAS project has successfully established robust technological and methodological platforms essential for dissecting the complexities of cardiac mechanotransduction and alternative splicing. Significant progress includes the development of engineered heart tissues (EHTs) derived from hiPSCs, optimized under physiological hypoxic conditions to closely mimic in vivo cardiac environments. Advanced single-cell sequencing has provided detailed insights into cardiomyocyte heterogeneity, revealing distinct cellular subpopulations that respond differently to mechanical stimuli and hypoxia.
Innovative multi-omics tools, including targeted single-cell phenomics, proximity-based proteomics (BioID), and advanced imaging technologies, have enabled comprehensive profiling of sarcomeric proteomes under defined mechanical conditions. These approaches facilitated the detailed analysis of splice factor localization and activity, uncovering novel mechanisms by which mechanical signals alter gene splicing. Notably, we characterized previously unknown RBM20 isoforms, highlighting their regulatory roles in cardiac development and disease.
Additional significant achievements include generating unique mouse models to study titin elongation and its impact on the splicing machinery, revealing adaptive isoform-specific regulation rather than broad transcriptional responses. The establishment of laser capture microdissection combined with full-length RNA sequencing provided unprecedented spatial resolution of gene expression and splicing patterns within heart tissues. These technical and scientific advancements have laid a strong foundation for targeted therapeutic strategies against heart failure.
Innovative multi-omics tools, including targeted single-cell phenomics, proximity-based proteomics (BioID), and advanced imaging technologies, have enabled comprehensive profiling of sarcomeric proteomes under defined mechanical conditions. These approaches facilitated the detailed analysis of splice factor localization and activity, uncovering novel mechanisms by which mechanical signals alter gene splicing. Notably, we characterized previously unknown RBM20 isoforms, highlighting their regulatory roles in cardiac development and disease.
Additional significant achievements include generating unique mouse models to study titin elongation and its impact on the splicing machinery, revealing adaptive isoform-specific regulation rather than broad transcriptional responses. The establishment of laser capture microdissection combined with full-length RNA sequencing provided unprecedented spatial resolution of gene expression and splicing patterns within heart tissues. These technical and scientific advancements have laid a strong foundation for targeted therapeutic strategies against heart failure.
MERAS has significantly advanced the understanding of how mechanical signals regulate cardiac splicing, revealing insights into regulatory networks previously unexplored. The development of single-cell and spatial multi-omics approaches provides a deeper resolution of cardiomyocyte heterogeneity and the transcriptional adaptations underpinning heart function and dysfunction.
The identification and functional validation of novel splice isoforms, particularly those regulated by RBM20, offer promising new targets for therapeutic intervention in heart diseases, especially HFpEF and dilated cardiomyopathy. MERAS’s results underscore the potential for splice modulation therapies, an area currently underexplored in clinical cardiology.
To ensure translation and prepare and clinical validation, we continue to improve the development of splice-modulating drugs and delivery systems for targeted therapeutics. Further research collaborations, supportive regulatory frameworks, intellectual property rights management, and strategic access to commercial and financial resources will be crucial for the successful commercialization and implementation of novel therapeutic strategies derived from our research.
The identification and functional validation of novel splice isoforms, particularly those regulated by RBM20, offer promising new targets for therapeutic intervention in heart diseases, especially HFpEF and dilated cardiomyopathy. MERAS’s results underscore the potential for splice modulation therapies, an area currently underexplored in clinical cardiology.
To ensure translation and prepare and clinical validation, we continue to improve the development of splice-modulating drugs and delivery systems for targeted therapeutics. Further research collaborations, supportive regulatory frameworks, intellectual property rights management, and strategic access to commercial and financial resources will be crucial for the successful commercialization and implementation of novel therapeutic strategies derived from our research.