Studies About Effect Of PFKFB3 Inhibition On The Differentiation Of CD4~+T Cells And Alleviation Of Experimental Autoimmune Encephalomyelitis

Objective:Multiple sclerosis(MS)is a chronic disease that causes nontraumatic nerve dysfunction in the central nervous system(CNS).It mainly occurs in young adults aged 20-40 years old,and the incidence is increasing year by year.The etiology and pathology of MS have not yet been fully understood,even though years of studies have been spent on MS.And also there is no effective medicine to treat MS once and for all.This study focused on studying the pathology of MS further and finds a novel strategy for its treatment.Content:Inflammatory lesions and demyelinating lesions are the main pathological indications of MS,and CD4~+T cells are considered to be the main pathogenic cells.Metabolic reactions within the cells control their proliferation,activation and even death.As the major pathogenic cells of MS,CD4~+T cells have undergone significant metabolic changes during the activation process.Using the mouse model of MS---experimental autoimmune encephalomyelitis(EAE),we first detected the expression of PFKFB3,which is a key enzyme during glycolysis,in pathological CD4~+T cells and studied its relationship with EAE development.Then,through comparing the cellularity of different CD4~+T cell subsets,we made a hypothesis that PFKFB3 might regulate the differentiation of CD4~+T cells.Finally,we conducted in vitro induction of Th1,Th17 and Treg cells to verify what we had found in vivo and further explored related molecular mechanism.Methods:PFKFB3 has been demonstrated to be key to various diseases with immune disorders.Using a mouse model of MS---experimental autoimmune encephalomyelitis(EAE),we studied the effect of PFK15-mediated PFKFB3inhibition on the survival,clinical and pathological score of the disease mice.In the meantime,the role of PFKFB3-mediated glycolysis in the differentiation of CD4~+T cells and its role in EAE disease development were explored and related molecular and cellular mechanisms were analyzed by western blot,real-time PCR,flow cytometry analysis,glucose absorption and lactate concentration detection.Results:In our study,we found an overexpression of PFKFB3 in EAE mice.Inhibition of PFKFB3 by PFK15 could alleviate the development of EAE,with improved survival and reduced clinical score.Through both in vitro and in vivo studies,we found that PFK15 could inhibit the differentiation of Th1 and Th17 cells,while promote the Treg differentiation,rebuilding the Th1/Th17/Treg balance in EAE mice and improving their survival.We also found that the role of PFK15 on Th1/Th17/Treg balance might dependent on the expression of mTOR.Conclusion:Collectively,our study demonstrated that PFKFB3 contributes to the development of EAE and targeted inhibition of PFKFB3 could regulate the balance of Th1/Th17/Treg and further alleviate EAE development,suggesting it as a potential target of MS treatment.