Project description:Eltrombopag, a small molecule thrombopoietin receptor (TPO-R) agonist and potent intracellular iron chelator, has shown remarkable efficacy to stimulate sustained multilineage hematopoiesis in patients with bone marrow failure syndromes, suggesting an effect at the most immature hematopoietic stem and multipotent progenitor level. While the functional and molecular effects of Eltrombopag on megakaryopoiesis have been carefully studied in the recent past, insights into its mechanistic impact on the earliest stages of hematopoiesis have been limited. Additionally, previous studies also revealed molecular pathway of Eltrombopag apart from stimulation of TPO signaling, detail characterization remains to be addressed. In this study, we investigated the effects of Eltrombopag, in comparison with TPO, on highly-purified primary hematopoietic stem cells (HSCs) from healthy human donors. The binding of Eltrombopag to TPO-R is species-specific to human and primate cells. we also utilized HSCs isolated mouse as separation-of-function models to directly assess the molecular effect of Eltrombopag on HSCs independent of TPOR stimulation.
Project description:Eltrombopag, a small molecule thrombopoietin receptor (TPO-R) agonist and potent intracellular iron chelator, has shown remarkable efficacy to stimulate sustained multilineage hematopoiesis in patients with bone marrow failure syndromes, suggesting an effect at the most immature hematopoietic stem and multipotent progenitor level. While the functional and molecular effects of Eltrombopag on megakaryopoiesis have been carefully studied in the recent past, insights into its mechanistic impact on the earliest stages of hematopoiesis have been limited. Additionally, previous studies also revealed molecular pathway of Eltrombopag apart from stimulation of TPO signaling, detail characterization remains to be addressed. In this study, we investigated the effects of Eltrombopag, in comparison with TPO, on highly-purified primary hematopoietic stem cells (HSCs) from healthy human donors. The binding of Eltrombopag to TPO-R is species-specific to human and primate cells. we also utilized HSCs isolated mouse as separation-of-function models to directly assess the molecular effect of Eltrombopag on HSCs independent of TPOR stimulation.
Project description:Identification of differentially expressed genes upon treatment with Eltrombopag in HL60 cells. HL60 cells were untreated, or treated with 3ug/ml of Eltrombopag for 36 hrs in RPMI with 10% FBS HL60 cells were untreated, or treated with 3ug/ml of Eltrombopag for 36 hrs in RPMI with 10% FBS
Project description:Identification of differentially expressed genes upon treatment with Eltrombopag in HL60 cells. HL60 cells were untreated, or treated with 3ug/ml of Eltrombopag for 36 hrs in RPMI with 10% FBS
Project description:Eltrombopag (EP) is a small molecule that acts on hematopoietic cells and megakaryocytes to stimulate the hematopoiesis. Mesenchymal stem/stromal cells (MSCs) are key hematopoietic niche regulators. In the current work we aimed to assess if EP exert any effect on MSCs function, especially on their hematopoietic-supporting ability, and if so, what are the changes (including transcriptomic) induced in MSCs after EP treatment. For this purpose, MSCs were isolated from 12 healthy donors and treated with 15 uM and 50 uM of EP for 24 hours. The toxicity of the drug on MSCs and their differentiation ability were analyzed, as well as the transcriptomic profile, ROS and DNA damage and finally the changes induced in the clonogenic capacity of HSC. We observed that EP acts on MSCs decreasing their adipogenic differentiation, increasing the expression of genes involved in hypoxia and other pathways and enhancing their ability to support hematopoiesis. We used high density oligo microarrays (Affymetrix Clariom_S_Human) to obtain the global programme of gene expression underlying the effect of the Eltrombopag (EP) and Thrombopoietin (TPO) in human MSCs (derived from bone marrow) and identify the genes activity.