OrganoidsFHeartbreak · Organoids For Heartbreak

Organoids For Heartbreak

Context and Overall Objectives of the Project Cardiovascular disease remains the leading cause of mortality in Europe, with post-injury cardiac remodelling and fibrosis representing major contributors to long-term morbidity and death. Current preclinical models, predominantly 2D monocultures or small-animal systems fail to capture the cellular diversity, mechanical environment, and complex signaling interactions of the human heart. As a result, approximately 90% of drug candidates fail in early clinical testing, underscoring an urgent need for physiologically relevant human in vitro models of cardiac fibrosis that can reliably predict therapeutic responses. The OrganoidsFHeartbreak project was conceived to address these unmet needs by developing an advanced human induced pluripotent stem cell (hiPSC)–based, three-cell-type spheroid model capable of reproducing the molecular and functional hallmarks of cardiac fibrosis. The scientific rationale rests on the critical role of fibroblast activation—driven by TGF-β and other pro-fibrotic cues, extracellular matrix accumulation, tissue stiffening, and progressive loss of cardiac function. A robust and inducible fibrosis model is therefore essential for understanding pathological mechanisms and for enabling early-stage drug discovery. The overall objective of the project was to establish a reproducible, physiologically relevant 3D cardiac organoid platform integrating hiPSC-derived cardiomyocytes, endothelial cells, and fibroblasts, and to validate its suitability for automated, high-content analysis of fibrosis initiation and inhibition. Specific goals included: 1. Generating and optimising the derivation of all three cardiac cell types, including development of a new differentiation strategy to obtain quiescent, stimulus-responsive hiPSC-derived fibroblasts. 2. Assembling multicellular cardiac spheroids with defined size and cell ratios, and demonstrating their structural integrity and lineage composition. 3. Inducing fibrosis with canonical activators (TGF-β1, Ang II) and quantifying fibrotic marker expression at both protein and transcript levels. 4. Integrating high-throughput kinetic image cytometry with customised CellProfiler pipelines to establish an automated, unbiased fibrosis-detection system. 5. Assessing functional consequences of fibrosis on contractility and calcium handling to confirm that the organoids reproduce clinically relevant aspects of cardiac dysfunction. Project Pathway to Impact The project positioned itself at the intersection of regenerative medicine, drug discovery, and advanced in vitro modelling. By overcoming major technical limitations—most critically, the generation of quiescent and activatable fibroblasts—the work produced a validated human 3D fibrosis model ready for further optimisation toward large-scale screening applications. The integration of automated imaging with quantitative image-analysis pipelines demonstrates that fibrotic activation and inhibition can be reliably detected across spheroids, establishing a methodological framework directly transferable to pharmaceutical screening workflows. The project’s pathway to impact is therefore threefold: Scientific impact: Provides a high-fidelity human model to study early mechanisms of cardiac fibrosis. Offers a platform for testing targeted therapies that modulate fibroblast activation and extracellular-matrix dynamics. Enables multi-marker, multiparametric readouts compatible with modern high-content screening technologies. Technological impact: Delivers an automated, reproducible workflow combining organoid culture, kinetic imaging, and computational analysis. Establishes protocols adaptable to 96- and 384-well formats, a prerequisite for high-throughput drug discovery pipelines. Demonstrates functional readouts (contractility, calcium flux) that complement structural fibrosis markers. Translational and societal impact: Enhances early-stage identification of promising anti-fibrotic compounds, accelerating development pipelines. Reduces reliance on animal models by providing a reliable human-based alternative. Contributes to EU priorities in precision medicine and improved cardiovascular health outcomes. In summary, the OrganoidsFHeartbreak project successfully lays the scientific and technological foundation for a next-generation human cardiac fibrosis platform. It addresses clear biomedical and translational needs and establishes a credible pathway toward impactful future applications in drug discovery, personalised medicine, and mechanistic research.

Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз