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Mechanisms regulating cardiovascular progenitor specification and migration

par Chiapparo, Giuseppe
Président du jury Parmentier, Marc
Promoteur Blanpain, Cédric
Publication Non publié, 2015-12-11

Thèse de doctorat

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Résumé :

During embryonic development and embryonic stem (ES) cell differentiation, the different cardiovascular cell lineages that compose the mature heart arise from different groups of cardiovascular progenitors (CPs), which contribute to the formation of different heart regions. Mesp1 expression is the earliest sign of cardiovascular development. Tracing of the early Mesp1-exressing cells reveals that all cardiovascular cell of the mature heart arise from Mesp1-derived cells. Various functional studies demonstrated that Mesp1 acts as a central regulator of cardiovascular lineage commitment, controlling the expression of early cardiac transcription factors, and promoting the differentiation into cardiomyocytes (CMs), endothelial cells (ECs) and smooth muscle cells (SMCs). In the first part of my thesis work, we generated a Mesp1-GFP reporter ES cell line allowing tracking and isolating the earliest Mesp1-expressing cells. We found that Mesp1 marks an early population of CPs that presents the ability to differentiate into CM, EC and SMC from both heart fields. Transcriptional profiling of Mesp1-CPs uncovered cell surface markers allowing their isolation. In this work, we demonstrated that Mesp1 is required for the specification of these early CPs. To ensure harmonious cardiovascular development, CP specification and migration must be tightly coordinated. In addition to the induction of cardiovascular cell fate, Mesp1 seems also involved in the migration of early CPs, as suggested by the cardiac malformation associated with the persistence of cardiovascular specification and differentiation in Mesp1 null embryo. The upregulation of the expression of Mesp2, its closest homologue, in the cardiac mesoderm, suggest that Mesp2 could partially compensate for Mesp1 function during early cardiogenesis, while Mesp1 presents a unique and non-redundant function during CP migration. However, Mesp2 KO embryos do not present cardiac malformation but rather major skeletal malformations. Inactivation of both Mesp1 and Mesp2 leads to the complete absence of mesoderm formation and consequently heart development, precluding the assessment of the redundant function of Mesp1 and Mesp2 during cardiovascular specification and differentiation. The second aim of this work was to investigate the mechanisms that compensate for Mesp1 functions during the early steps of cardiogenesis, and those that control the unique Mesp1 function during CP migration. Using inducible gain of function experiments during ES cell differentiation, we found that Mesp2 was as efficient as Mesp1 at promoting CP specification, EMT and cardiovascular differentiation. However, only Mesp1 stimulated the polarity and directional cell migration through a cellular autonomous mechanism. Transcriptional analysis and ChIP experiments revealed that Mesp1 and Mesp2 activate a common set of target genes that promote CP specification and differentiation. We identified two direct Mesp1 target genes, Prickle1 and RasGRP3, that are strongly induced by Mesp1 and not Mesp2 and which control the polarity and the speed of cell migration. Altogether our results identify the molecular interface controlled by Mesp1 that links CP specification and migration during ES cell differentiation.