| ObjectiveGiant cell tumor of bone is one of the most common bone tumors in Asian population,and it can damage bone tissue,disability rate,recurrence and certain metastasis.Therefore,it is focus to find the mechanism of bone damage and therapeutic targets.The most studies are about surgical exploration or retrospective research,and basic research are on signaling pathway,so we focus on the cell types to analyze the cellular heterogeneity.In combination with gene sequencing technology,the separation of multinuclear osteoclast-like giant cells and osteoclasts that cause bone erosion was realized,and the different genes of the them carried out and mechanisms for the diagnosis,treatment,prognosis assessment,invasion and metastasis of GCTB.MethodsThe method is single cell sequencing.It is developed from the second generation sequencing technology,and it can achieve the separation of cell types,independent sequencing and gene library.It can also improve the precision of the gene expression.This method can truly achieve the resolution of cell heterogeneity,which has opened up a new direction for the research of tumor development,diagnosis and treatment.In this study,GCTB tissue was processed by processing-dissociation-filter detection until sequence and build a database.The data was subjected to standardized filtering,transformation,dimensionality reduction and other processing to classify cells into 12 groups.We used the two-way query definition method when defining the types,extracting differentially expressed genes and querying information,collecting marker genes and then comparing,twelve cell populations are defined.This fills the gap in defining cell methods.After identifying the osteoclast-like giant cell and the osteoclast,we extracted the highly expressed genes of these two,and annotated them with GO(molecular and organisms function)to search for genes related to the excessive bone resorption.ResultThe cell types include: NKT cells,osteoblasts,plasma cells,B cells,giant cells,macrophages,T cells,stromal cells,monocytes,osteoclasts,a small number of muscle cells,osteoclast precursor cells,vascular endothelial cells and fibroblasts.50 highly expressed in osteoclast-like giant cells and osteoclasts genes were picked.More than half(27)are the overlapping parts in the two cell groups by Venn.After performing heat map analysis of these genes,it was found that genes higher express in osteoclasts include CA2,ATP6V1E1,CKB,SPP1,ATP6V1C1,ATP6V1G1,GLRX,CSTB,ATP6V0E1,MATK,RAC2,SLC9B2,ATP6V1B2,CHCHD10,TCIRG1,ATP6V1H,ATP6V1F,RALA,ACP5,CTSD,CD68,genes highly expressed by giant cells include TKT,SNX10,MMP9,CD84,RGS10,CTSK.Upon the GO annotation for each gene,we found that: in addition to positively regulating bone resorption,the gene SPP1 also participates in the process of osteoblast differentiation and ossification,but the expression of SPP1 in giant cells is lower.The gene MATK is involved in the ErbB2 signaling pathway,and ErbB2 has been reported in research overexpression accompanied by lung metastases in patients,and it also marked a poor prognosis.Among the genes highly expressed in giant cells,the gene CTSK participates in negatively regulate intraosseous ossification,bone resorption and cartilage development.These may cause the destruction of GCTB,which has also been reported in research: AKT / NFATc1 / CTSK axis promotes osteoclast genesis and bone resorption of the skull,while the mechanism in GCTB has not been elucidated.Gene SNX10’s functions include many osteoclast-like functions,and studies have shown SNX10 is involved in osteoclast activation.The mutation of SNX10 will cause osteoclast dysfunction,and the mechanism in GCTB has not been elucidated.ConclusionIn this study,we achieve the analysis of the single cell level and gene level of GCTB,defined the cell type and genes SPP1,MATK,CTSK,and SNX10,which can cause tumor bone erosion characteristics,were identified to make up for the shortcomings in the current research field. |
PDF Full Text Request