LYL1 (Lymphoblastic leukemia-derived sequence 1) est un facteur de transcription basic hélice-boucle-hélice (bHLH) exprimé dans les lymphocytes B, les cellules myéloïdes et les cellules endothéliales (CE). Les souris déficientes pour Lyl1 (Lyl1-/-) sont viables et chez la souris adulte, LYL1 a un rôle majeur dans la maturation des vaisseaux sanguins nouvellement formés et dans le contrôle de la perméabilité vasculaire basale, suggérant l'importance de LYL1 dans le maintien de la quiescence et/ou stabilisation des CE. Les vaisseaux sanguins représentent une barrière entre le sang et le tissu conjonctif. Ils peuvent également jouer le rôle de niche vasculaire contenant des progéniteurs des différentes cellules murines (par exemple, des cellules hématopoïétiques, des cellules β-pancréatiques, des cellules neuronales, des cellules hépatiques et des cellules adipeuses). Les deux tissus adipeux, blancs et bruns (WAT et BAT), sont très vascularisés. Jusqu'à présent, rien n'était connu sur le rôle de LYL1 dans le tissu adipeux. Les résultats présentés dans cette thèse montrent que l'augmentation significative du poids corporel des mâles Lyl1-/- par rapport aux souris sauvages (WT), sous régime normal, n'est pas associée à des troubles métaboliques. Ils présentent également un poids plus élevé de tissus adipeux (WAT et BAT) et de plus grandes gouttelettes lipidiques. In vivo, la perte de Lyl1 accélère le processus de différenciation des cellules souches adipeuses (CSA), puisque les adipocytes blancs et bruns sont matures et actifs plus tôt. De plus, les CSA sont moins nombreuses dans les tissus adipeux, ce qui confirme que la perte de Lyl1 favorise la différenciation des CSA vers adipocytes matures. Nous avons également démontré que Lyl1 est exprimée dans les CSA et les pré-adipocytes, suggérant un rôle direct dans LYL1 dans la différenciation adipocytaire. D'autre part, les vaisseaux des WAT des souris Lyl1-/- sont mal recouverts de cellules murales et plus perméables, suggérant que la niche vasculaire des tissus adipeux pourrait être perturbée. Sous alimentation riche en graisses (HFD), le poids corporel et le poids du tissu adipeux sont plus faibles chez les souris Lyl1-/- par rapport à WT. De plus les souris Lyl1-/- présentent de plus petites gouttelettes lipidiques que les WT, sous HFD. Ces résultats préliminaires, suggèrent que les souris Lyl1-/- pourraient être protégées contre l'obésité induite par l'alimentation. Cependant d'autres expériences sont nécessaires pour valider ces résultats. Il existe probablement un mécanisme de compensation qui se met en place chez les souris Lyl1-/- sous HFD. Ce travail a démontré que, sans Lyl1, la différenciation adipocytaire est accélérée et que la niche vasculaire adipocytaire est perturbée. / LYL1 (Lymphoblastic leukemia-derived sequence 1) is a basic helix-loop-helix (bHLH) transcriptional factor, which is expressed in B lymphocytes, myeloid cells and endothelial cells (EC). Lyl1 deficient (Lyl1-/-) mice are viable and in adult mice, LYL1 has an active role in the maturation of newly formed blood vessels and is also involved in the control of basal vascular permeability, suggesting that LYL1 is required for the maintenance of EC quiescence and stabilization. Blood vessels provide a barrier between connective tissue and blood. They also have been described as “vascular niche” containing progenitors of different murine cells (e.g. hematopoietic cells, pancreatic β-cells, neuronal cells, liver cells and adipose cells). Both white and brown adipose tissues (WAT and BAT) are highly vascularized. Up to now, nothing was known concerning the role of LYL1 in adipose tissue. The results presented in this thesis revealed that the significant increase in body weight of Lyl1-/- males compared to their wild type (WT) littermates under chow diet is not due to any metabolic disorders. They also showed higher adipose tissue weights (BAT and WAT) and bigger lipid droplets. In vivo Lyl1 deficiency cause early differentiation process of adipose stem cells (ASCs) since both white and brown adipocytes are mature and active faster. In addition, ASCs are less numerous in Lyl1-/- adipose tissues, which confirm that Lyl1 deficiency favors the differentiation of ASCs towards mature adipocytes. We also demonstrated that Lyl1 is expressed both in ASCs and pre-adipocytes, suggesting a direct role of LYL1 in adipocyte differentiation. On the other hand, the vessels in Lyl1-/- WAT are poorly covered with mural cells and more permeable, proposing that adipose stem cell vascular niche could be disturbed. Under high fat diet (HFD), total body weight and adipose tissue weight are lower in Lyl1-/- mice compared to WT. Moreover smaller lipid droplets were observed in Lyl1-/- mice under HFD. These preliminary results suggest that Lyl1-/- mice could be protected from diet-induced obesity. However more experiments are needed to validate these results. Probably there is a compensatory type of mechanism going on under HFD in Lyl1-/- mice. This work demonstrated that under Lyl1 deficiency adipocyte differentiation process becomes faster and adipose tissue vascular niche could be disturbed.

Umbilical cord blood (UCB) is used as a source of haematopoietic stem cells (HSCs) for transplantation but shows defective homing to the bone marrow niche and delayed haematological reconstitution. Following transplantation, HSCs will home to the bone marrow in response to the CXCL12 chemokine, adhere to the bone marrow sinusoidal endothelial cells and then migrate into and lodge in bone marrow niches. In addition to CXCR4, a variety of molecules have been described as being important in these processes. In this laboratory, junctional adhesion molecule-A (JAM-A) was shown to be expressed on human UCB CD133⁺/CD34⁺ cells and regulated by hypoxia. In this thesis, further phenotypic studies show that this molecule is most highly expressed on human CD41a⁺ megakaryocytes and CD14⁺ monocytes/macrophages in UCB. JAM-A was also found to be expressed on all human UCB CD133⁺ cells, which have been shown by others to encompass the HSCs and early myeloid-lymphoid precursors and on the majority of CD34⁺ haematopoietic progenitor cells (HPCs). While it is also present on bone marrow sinusoidal endothelium (BMEC), JAM-A is not detected on cultured bone marrow mesenchymal stromal cells (MSCs). JAM-A blockade, silencing and overexpression experiments showed that JAM-A contributes to, but is not solely responsible for, the adhesion of CD34⁺ haematopoietic progenitor cells to IL-1β activated BMEC-60 cells and fibronectin. Lack of significance in cell migration suggested that JAM-A is more likely to act as an adhesion molecule or a regulator of adhesion rather than as a migratory molecule in such cells. Further functional studies using the proximity ligation assay highlight a potential association of JAM-A with CXCR4 and the adhesion molecules, tetraspanin CD82 and integrin β1. Mechanistic studies were commenced to establish if JAMA could modulate CXCR4 signalling following CXCL12 stimulation, but time constraints prevented these from being completed. These preliminary experiments which were carried out first in the Jurkat cell line lacking JAM-A or transduced to express JAM-A, however, suggest that JAM-A may modulate CXCL12-induced Rap1 phosphorylation and ERK1/2 phosphorylation. The former pathway is important for integrin function and the latter pathway is important in cell adhesion. The results described here, although requiring finalisation, support the hypothesis that JAM-A acts as an adhesion molecule and also may fine tune CXCR4 and integrin mediated functions on human CD34⁺ cells, thereby potentially regulating engraftment of these cells to the bone marrow niche.