Study On ACT001 Effects In Expression Pattern Changing As Well As Invasion Inhibiting Via CCR2 In GBM

Introduction:Glioblastoma(GBM)is the most common primary cancer of the central nervous system(CNS),which is always in form of highly invasive growth.The median survival of GBM patients is only 14.6 months.ACT001,as a promising drug for glioma,has a unique character with ability to easily pass through blood brain barrier.As a derivative of immunomodulator,ACT001 has a unique effect on immunogenic cells.However,as the Phase I clinical trials of ACT001 in Australia goes better,the mechanism in treating with glioma is still unclear.Our research aims to reveal the role of ACT001 in the treatment of gliomas via high-throughput screening followed by bioinformatics research and immunological research.This research will reveal the mechanism of ACT001 treating with gliomas in some degree,and combination of temozolomide(TMZ)in the treatment for tumor-bearing mice that is expected to provide a theoretical basis for the future combined-application of temozolomide.Methods:In this study,gene expression pattern analysis when treating with glioma stem cell,glioma cell,tumor-associated macrophages,GL261 tumor-bearing mice,and combined with temozolomide in the treatment of GL261 tumor-bearing mice was performed with annotation databases GO,KEGG,Reactome,MSig Db with differential expressed genes.The enrichment analysis and gene set enrichment analysis were performed to identify and screen the whole gene expression patterns.Finally,we screened CCR2 as the key protein in glioma invasion and migration under treatment of ACT001.Results:Data from micro array shows that the stemness of glioma stem cells can be inhibited by ACT001.Gene expressed pattern changed in induced M2 macrophage shows that cell phenotype was transformed into epithelial by ACT001.Data from glioma cells suggests that ACT001 can kill glioma cells but is not as good as TMZ.Micro array of tumor-bearing mice shows that chemokine-related pathways were inhibited by ACT001,especially CCR2.Data from flow cytometry shows glioma induced-recruitment of myeloid-derived suppressor cells(MDSCs)was inhibited by ACT001.Western Blot results suggest that tumor metastasis niche has been inhibited by ACT001 via CCR2 pathway,which can inhibit invasion and migration of glioma cells.Conclusions:ACT001 can inhibit recruitment of MDSCs by inhibiting CCR2 and can reverse the tumor induced immune suppression in the tumor microenvironment.ACT001 can also inhibit the formation of pre-metastasis niche by CCR2 and MDSCs,which can inhibit the invasion and migration of glioma cells.This excited result shows that ACT001 would have a good outcome combined with TMZ in treating glioma.