Dissecting The Underlying Mechanism Of GM-CSF/G-CSF Induced Myeloid Progenitor Cell Differentiation

Purpose: The myeloid progenitor cells derived from hematopoietic stem cells can eventually differentiate into a series of mature blood cell components,including mononuclear macrophages.The function of macrophages is crucial to the maintenance of homeostasis.Conversely,defective regulation of myeloid differentiation will lead to immune deficiency.Studies on the differentiation of macrophages provide the basis for the treatment of blood diseases.The purpose of this study is to explore the main mechanisms of the macrophages differentiation.Research content: The research on granulocyte-macrophage colony stimulating factor(GM-CSF)induced macrophage differentiation,the influence of m TOR signaling pathways on the macrophage differentiation,and the effect of autophagy pathway which situate the downstream of m TOR in macrophages differentiation.Methods: Flow cytometry and quantitative PCR were used to determine macrophage differentiation model.By pharmacological and genetic modification to activate or inhibit m TOR signaling and autophagy activity,the differentiation related markers,m TOR signaling and autophagy related components were tested with western blot,immunofluorescence and other technical.Results: GM-CSF triggers macrophage differentiation and activation of the m TOR signaling pathway.Activation or inhibition of m TOR signaling enhances or attenuates macrophage differentiation,respectively.We further showed that autophagy is inhibited with the addition of GM-CSF.Furthermore,pharmacological inhibition and genetic modification of autophagy enhances macrophage differentiation and rescues the inhibitory effect on differentiation caused by m TOR inhibition.Conclusion: The m TOR signaling pathway regulates macrophage differentiation of myeloid progenitors by inhibiting autophagyPurpose: Neutrophils are derived from myeloid progenitor cells,playing an important role in the innate immune system.Aberrant neutrophils(amount or functions)can result in many kinds of blood diseases.Dissecting the underlying mechanismof the differential process from the myeloid progenitor cells to the neutrophil is the key point to study how to prevent and treat the blood disease.The purpose of this study is to explore the relevant molecular and signaling pathways of neutrophil differentiation,to provide new targets for hematopoietic differentiation research and will shade new insight on the research of hematological diseases.Research content: Study the role of NLRP12 in the neutrophil differentiation induced by granulocyte colony stimulating factor(G-CSF).Figure out the relationship between NLRP12 expression and ?l?gthe activity of non-canonical NF-?B signal pathway and ERK1/2 pathways.Methods: Using flow cytometry and quantitative PCR to confirm the differentiation model.Western blot and quantitative PCR were used to confirm the interference efficiency of NLRP12,which was knocked down by two sh-RNAs.Detecting the changes of key moleculatr of non-canonical NF-?B signal pathway,ERK1/2 pathways and markers of neutrophilResults: In the process of G-CSF induced neutrophil differentiation,the expression of NLRP12 increased significantly.The absence of NLRP12 expression inhibits the non-canonical NF-?B signaling pathway,but activates the Erk1/2 pathway,which inhibited the neutrophil differentiation.Conclusions: NLRP12 promotes mouse neutrophil differentiation through regulation of Non-canonical NF-?B and MAPK ERK1/2 Signaling...