NSD2 Expression In Prostate Cancer And Correlation With Immune Infiltrating Cells

Objective:To investigate the expression of NSD2,CD4~+T and CD8~+T cells in prostate cancer(PCa)and their effects on bone metastasis and prognosis of prostate cancer;Further,illustrate the involvement of NSD2 in the infiltration of immune cells and the potential molecular mechanisms in prostate cancer.Methods:Immunohistochemistry was performed to evaluate the expression pattern of NSD2 in benign prostatic hyperplasia(BPH),prostatic intraepithelial neoplasia(PIN),PCa and metastatic castration resistant prostatic cancer(m CRPC).Meanwhile,immunohistochemically labeled PCa with CD4~+and CD8~+T and counted,and divided into high and low expression groups according to the percentage/median number of NSD2,CD4,CD8 positive cells,and analyzed the correlation between the three and their correlation with bone metastasis and prognosis.Furthermore,using Single-cell RNA sequencing data,then gene set enrichment analysis(GSEA)was used to correlate NSD2 with certain downstream pathways.We used the Immuno-Oncology-Biological-Research(IOBR) software package to analyze the potential roles of NSD2 in the tumor microenvironment.Results:(1)The positive expression rate of NSD2 in BPH,PIN,PCa and m CRPC increased gradually.(2)The positive rate of NSD2 in PCa was 91.2%(52/57),and the positive rates of CD4~+T and CD8~+T in PCa were both 100%(57/57).NSD2 was highly expressed in 31.6%(18/57)of cases,and the high expression rates of CD4~+T and CD8~+T were 22.8%(13/57)and 45.6%(26/57),respectively.(3)Plasma PSA and CD4~+TILs were independent risk factors for PCa bone metastasis(P<0.05).(4)Patients with high NSD2 expression had shorter OS than those with low NSD2 expression(OS:P=0.007),and multifactorial Cox regression analysis showed that patients with PCa with higher Gleason score had shorter survival(OS:P=0.023).(5)High NSD2 expression was positively correlated with CD4~+TILs(P=0.0393)and negatively correlated with CD8~+TILs(P=0.0479),and NSD2 expression was correlated with prognosis of PCa patients(P=0.0007).A novel immune classification based on NSD2 expression and CD4~+TILs(P=0.0002)and CD8~+TILs(P=0.0102)successfully stratified the OS of PCa.(6)GSEA enrichment analysis showed that NSD2-KO gene was associated with IL-2-STAT5signaling pathway(P=0.01),IFN-γsignaling pathway(P=0.02),TGF-βsignaling pathway(P=0.01)and DNA repair pathways(P=0.01).(7)IOBR analysis showed that NSD2-KO was not significantly correlated with the immunosuppressive tumor microenvironment,but the infiltration density of follicular CD8~+T cells and CD4~+T cells was significantly higher in samples with NSD2-KO than in samples with CTR-KO,as calculated by CIBERSPOT.Conclusions:NSD2 expression is progressively increased in BPH,PIN,PCa and m CRPC with prostate cancer development and progression and is a risk factor for bone metastasis and poor prognosis in prostate cancer.Notably,NSD2 regulates the infiltration of CD4~+T and CD8~+T cells in prostate cancer and is important for the immune microenvironment.NSD2 is associated with multiple pathways involved in both intrinsic and acquired immunity,and NSD2 is expected to be a new target for immunotherapy.