Biomarker Screening And Verification Of Multiple Myeloma Treated By Acupuncture Combined With Medicine

Background and ObjectiveMultiple myeloma(MM)is a malignant tumor which causes proliferation of abnormal plasma cells in the bone marrow.MM has the characteristics of high relapse and incurability.Although there are a variety of clinical treatment strategies and new drugs,MM is fraught with great difficulties in therapy.Acupuncture is the world’s intangible cultural heritage.It has long been emphasized in the Neijing that acupuncture can calm the viscera,flat Yin and Yang,and smooth the meridian patency.Many studies have reported that acupuncture has potential therapeutic effects on cancer symptoms such as pain,vomiting,xerostomia,hot flashes,and fatigue.Therefore,the combination of acupuncture and medicine treatment of multiple myeloma can alleviate patient symptoms and improve the quality of life.The purpose of this study is to explore effect and possible mechanism of acupuncture combined with medicine treatment of multiple myeloma.This research identifies the serum metabolomics characteristics of the MM animal models treated with the combination of acupuncture and drug,its superior metabolic pathway and target genes,and studies further about the influence of the pathway on MM.This research finds potential biomarkers and verified them.This study will supply theoretic support for acupuncture combined with medicine treatment of MM.Methods1.Training set and validation set of 5TMM3VT MM murine model are established.These mice are treated with acupuncture and bortezomib.Their blood is collected from caudal vein once a week.At the same time,the survival status of mice is recorded.2.In terms of untargeted metabolomics for qualitative analysis,LC/MS instrument is used to analyze the metabolic differences in murine serum.This research finds unique pathways and targets for acupuncture combined with bortezomib treatment of MM.3.In terms of targeted metabolomics for quantitative analysis,the murine serum levels of potential biomarkers are measured on a UHPLC-MS/MS instrument to verify the result of untargeted metabolomics.4.Combined the gene expression omnibus databases(GEO),the association analysis of gene differential expression and overall survival(OS)in different databases is applied to find and verify the potential target genes of acupuncture combined with bortezomib treatment of MM.Results1.Animal experiments in the training set and validation set show that compared with acupuncture group or bortezomib group,MM mice treated by acupuncture combined with bortezomib have a longer survival period.2.The results of untargeted metabolomics show that compared with other treatment groups,the serum levels of 32 metabolites are significant decreased in MM murine model treated by acupuncture combined with bortezomib.Pathway analysis indicate that the arginine and proline metabolism pathway involving arginine and ornithine reducing gets the highest pathway impact value score,and ornithine also participates in the regulation of glutathione metabolism pathway.3.Targeted metabolomics results confirm that the level of ornithine in MM murine model treated with combined acupuncture and bortezomib is lower than the disease control group.4.There are 3 genes encoding enzymes involved in the regulation of ornithine metabolism.GEO databases analysis of these genes expression suggests that MM patients with higher ODC1 expression are evidently associated with poor overall survival,and ODC1 expression increased with the progression of MM.ConclusionAcupuncture and medicine have synergistic effects in the treatment of MM.The ornithine metabolic pathway is the unique dominant metabolic pathway in the effect of acupuncture combined with bortezomib treatment of MM.Ornithine can be used as a biomarker for the combination of acupuncture and bortezomib therapy for MM.Higher ODC1 expression is evidently associated with poor prognosis in MM patients.ODC1 is a potential biological target for prognosis prediction of MM.