bioRxiv Science⌕ Search

Biology subjects

Lekkos, K.

Publications and source records attributed to Lekkos, K..

4 recordsLinked to original sources

A balanced circadian glycolytic rhythm drives cardiomyocyte cell cycle progression during fish heart regeneration.

The ability of heart tissue to repair itself after injury has fascinated scientists for decades1,2. Researchers have long studied the internal body clock, or circadian rhythm, for its role in coordinating daily cycles of metabolism and cell activity3,4, but its relevance to heart repair has remained unknown. This study explores, for the first time, whether natural daily rhythms influence heart regeneration--a process driven by cardiomyocyte proliferation. We discovered that DNA replication, mitosis, oxidative phosphorylation, and glycolysis follow a precise daily order in regenerating zebrafish hearts. Disrupting core clock gene expression abolishes the rhythms of glycolysis and mitosis, preventing cardiomyocyte cell cycle progression and regeneration. Insulin-resistant Astyanax mexicanus cavefish, which have adapted to dark caves, similarly show a loss of mitosis rhythm and cardiomyocyte cell cycle progression, which we find is caused by reduced glycolysis. Despite this reduction, glycolysis rhythm displays a larger amplitude in cavefish--a pattern recapitulated in insulin-resistant zebrafish. Insulin resistance resets metabolic rhythms to the morning, which is equally detrimental to regeneration. Here, we show that successful cardiac regeneration depends on synchronised clock and glucose rhythms, which together orchestrate the cell cycle events essential for cardiomyocyte proliferation and tissue repair.

physiology↗

Absence of a prolonged macrophage and B cell response inhibits heart regeneration in the Mexican cavefish

A balanced immune response after cardiac injury is crucial to successful heart regeneration, but knowledge of what distinguishes a regenerative from a scarring response is still limited. The Mexican cavefish provides a unique comparative model to study heart regeneration and scarring within a single species. Surface-dwelling fish are capable of heart regeneration whereas their cave-dwelling Pachon counterparts lack this ability, similar to the human heart. Using single-cell transcriptomics and immune perturbations, we find significant differences in the immune response between the two populations. Unlike the transient response in the scarring Pachon, the regenerative surface fish heart generates an unexpected functionally active prolonged innate and adaptive immune response at the late stages of regeneration. Inhibiting the overall prolonged immune response impairs regeneration and cardiomyocyte proliferation. Further characterisation of specific cell types shows that late-present macrophages are phagocytic, and their depletion disrupts regeneration but not cardiomyocyte proliferation while inhibiting B cells impairs regeneration by reducing cardiomyocyte proliferation. This B cell response is conserved in zebrafish. Our findings reveal critical immune mechanisms distinguishing regenerative and non-regenerative responses, offering insights for potential therapeutic strategies to enhance heart repair.

developmental biology↗

Endothelial Slit2 guides the Robo1-positive sympathetic innervation during heart development

Axon guidance cues direct nerves in the heart during development, disease and regeneration. These cues determine cardiac innervation patterning by regulating the balance between chemo-attraction and chemo-repulsion. However, the role of one of the most crucial ligand-receptor combinations among axon guidance molecules, the Slit chemo-active ligands and their Roundabout (Robo) transmembrane receptors, remains unknown during cardiac innervation patterning. To test if Slit-Robo signalling is important for cardiac innervation guidance, we analysed Slit and Robo mouse knock-outs. Constitutive Slit2-/- ventricles showed significantly reduced innervation, while Slit3-/- hearts showed temporary increased levels of innervation compared to wild-type littermate controls. Whereas innervation was not affected in Robo2-/- mice, the phenotype seen in Slit2-/- ventricles was recapitulated in Robo1-/- mice. Detailed expression analysis identified expression of Slit2 ligand in the endothelium of the coronary vessels, while Slit3 was highly present in the coronary smooth muscle wall and in the innervation. Both the Robo1 and Robo2 receptors were present in the nerves and at low levels in the vessels. Knocking out Slit2 specifically in the endothelium recapitulated the defects seen in the constitutive Slit2-/- hearts. Ex vivo axon guidance cultures showed that attraction of axons extending from the ganglia was strongly reduced in ventricles with absence of endothelial Slit2 compared to wild-type controls. In absence of endothelial Slit2, adult mice showed reduced response to challenging the sympathetic innervation. In conclusion, we have identified an important new chemo-active Slit2-Robo1 pathway required for correct cardiac innervation development.

developmental biology↗

Injured endocardium obtains characteristics of haemogenic endothelium during adult zebrafish heart regeneration

Reactivation of embryonic developmental pathways during regeneration aims to restore tissue architecture and functionality. We previously reported that following cryoinjury, a heterogeneous population of Runx1-expressing endocardial cells differentially upregulates genes associate with scarring and myofibroblast identity. Further analysis of our published RNAseq data alongside 5 publicly available datasets now identifies additional heterogeneity in the Runx1-positive injured endocardium. Here, we show that the endocardium also reactivates a dormant endocardial-to-haematopoietic transition (EHT) mechanism. Runx1-expressing endocardial cells upregulate genes associated with haemogenesis and morphologically display features of EHT. Live imaging shows cells budding off the endocardium and lineage analysis identifies overlap with leukocyte markers. Ablation of runx1 function further shifts differentiation of the endocardium towards the EHT fate. The identification of transient runx1-expressing cells transitioning towards myofibroblast or haemogenic endocardium identities demonstrates the complexity of the zebrafish endocardial injury response and highlights the role of Runx1 in regulating cell fate decisions in the endocardium.

developmental biology↗