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| Item Type: | Article |
|---|---|
| Title: | Enhancing KLF15 activity in cardiomyocytes: a novel approach to prevent pathological reprogramming and fibrosis via nuclease-deficient dCas9VPR |
| Creators Name: | Schoger, Eric, Kim, Rosa, Bleckwedel, Federico, Peralta, Tomas, Priesmeier, Laura, Fischer, Janek, Stengel, Laura, Rocha, Cheila, Santos, Gabriela L., Lutz, Susanne, Boileau, Etienne, Baumgarten, Nina, Schulz, Marcel H., Dieterich, Christoph, Müller, Oliver J., Cyganek, Lukas, Cabrera-Orefice, Alfredo, Eberl, Hanna, Maack, Christoph, Streckfuss-Bömeke, Katrin, Pavez-Giani, Mario, Doroudgar, Shirin, Sossalla, Samuel T. and Zelarayán, Laura C. |
| Abstract: | Transcriptional activity perturbation holds promise for selectively modulating harmful transcriptional networks, but its therapeutic potential remains largely unexplored. We employed a network-based analysis of single-cell heart transcriptomes to identify transcription factor activities linked to pathological cardiomyocytes in vivo. This analysis revealed that transcriptional activity of Krüppel-like factor 15 (KLF15) exhibited the most significant change in pathological cardiomyocytes, characterized by less effective repression of disease-associated genes in stressed hearts, which correlated with reduced KLF15 expression. To restore KLF15 activity, we utilized CRISPR/nuclease-dead (d)Cas9-based transcriptional enhancement (CRISPRa) in cardiomyocytes, which effectively abolished fetal reprogramming by simultaneously suppressing pathological gene expression and restoring metabolic homeostasis under sustained stress conditions. Furthermore, we identified a novel cell-nonautonomous anti-fibrotic effect mediated by cardiomyocyte-fibroblast crosstalk, and revealed the contribution of KLF15-dependent Alpha-2-glycoprotein 1, zinc-binding (AZGP1) regulation in this process. We also elucidated the upstream mechanisms of KLF15 regulation, highlighting its role as a cell-specific downstream target of the broad TGF-β canonical signaling pathway, along with its downstream-dependent mechanisms in human cardiomyocytes. Finally, to enhance the therapeutic potential of this approach, we engineered and validated an adeno-associated viral (AAV) vector with a small CRISPRa system for endogenous regulation in human cardiomyocytes suitable for clinical applications. Overall, we elucidated a regulatory circuit involving TGF-β, KLF15, and AZGP1, which coordinates critical pathological responses through cellular crosstalk between cardiomyocytes and fibroblasts. Importantly, we demonstrated the efficacy of CRISPRa as an epigenetic intervention restoring a critical transcriptional function disrupted in non-genetic heart failure. This approach provides a promising blueprint for future adaptation targeting additional non-hereditary pathologies. |
| Keywords: | CRISPR-Cas Systems, Cardiac Myocytes, Cellular Reprogramming, Fibroblasts, Fibrosis, Gene Expression Regulation, Kruppel-Like Transcription Factors, Transcription Factors, Animals, Mice |
| Source: | Signal Transduction and Targeted Therapy |
| ISSN: | 2095-9907 |
| Publisher: | Springer Nature |
| Volume: | 11 |
| Number: | 1 |
| Page Range: | 76 |
| Date: | 3 March 2026 |
| Official Publication: | https://doi.org/10.1038/s41392-026-02593-9 |
| PubMed: | View item in PubMed |
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