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. 2021 Jun 2:2021:9967954.
doi: 10.1155/2021/9967954. eCollection 2021.

Construction and Validation of a Macrophage-Associated Risk Model for Predicting the Prognosis of Osteosarcoma

Kang-Wen Xiao  1 , Zhi-Bo Liu  1 , Zi-Hang Zeng  2 , Fei-Fei Yan  1 , Ling-Fei Xiao  1 , Jia-Li Li  2 , Lin Cai  1
Affiliations

Construction and Validation of a Macrophage-Associated Risk Model for Predicting the Prognosis of Osteosarcoma

Kang-Wen Xiao et al. J Oncol. .

Abstract

Background: Osteosarcoma is one of the most common bone tumors among children. Tumor-associated macrophages have been found to interact with tumor cells, secreting a variety of cytokines about tumor growth, metastasis, and prognosis. This study aimed to identify macrophage-associated genes (MAGs) signatures to predict the prognosis of osteosarcoma.

Methods: Totally 384 MAGs were collected from GSEA software C7: immunologic signature gene sets. Differential gene expression (DGE) analysis was performed between normal bone samples and osteosarcoma samples in GSE99671. Kaplan-Meier survival analysis was performed to identify prognostic MAGs in TARGET-OS. Decision curve analysis (DCA), nomogram, receiver operating characteristic (ROC), and survival curve analysis were further used to assess our risk model. All genes from TARGET-OS were used for gene set enrichment analysis (GSEA). Immune infiltration of osteosarcoma sample was calculated using CIBERSORT and ESTIMATE packages. The independent test data set GSE21257 from gene expression omnibus (GEO) was used to validate our risk model.

Results: 5 MAGs (MAP3K5, PML, WDR1, BAMBI, and GNPDA2) were screened based on protein-protein interaction (PPI), DGE, and survival analysis. A novel macrophage-associated risk model was constructed to predict a risk score based on multivariate Cox regression analysis. The high-risk group showed a worse prognosis of osteosarcoma (p < 0.001) while the low-risk group had higher immune and stromal scores. The risk score was identified as an independent prognostic factor for osteosarcoma. MAGs model for diagnosis of osteosarcoma had a better net clinical benefit based on DCA. The nomogram and ROC curve also effectively predicted the prognosis of osteosarcoma. Besides, the validation result was consistent with the result of TARGET-OS.

Conclusions: A novel macrophage-associated risk score to differentiate low- and high-risk groups of osteosarcoma was constructed based on integrative bioinformatics analysis. Macrophages might affect the prognosis of osteosarcoma through macrophage differentiation pathways and bring novel sights for the progression and prognosis of osteosarcoma.

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Conflict of interest statement

The authors declare that there are no conflicts of interest.

Figures

Figure 1
Figure 1
Flowchart of this study.
Figure 2
Figure 2
Identification of prognostic MAGs through PPI and DGE and immune infiltration of TARGET-OS: (a) PPI analysis of 384 MAGs, (b) differential gene expression analysis between osteosarcoma samples and normal bone sample, (c) correlation plot of each immune cell in TARGET-OS, and (d) immune infiltration of each sample in TARGET-OS.
Figure 3
Figure 3
The results of survival curve for 5 MAGs (BAMBI, PML, GNPDA2, WDR1, and MAP3K5) and the expression of each MAG in a normal bone sample and osteosarcoma: (a)–(e) survival curve of 5 MAGs (BAMBI, PML, GNPDA2, WDR1, and MAP3K5) by Kaplan–Meier method and (f)–(j) the boxplots of expression of each MAG in normal bone sample and osteosarcoma. ∗p < 0.05 and ∗∗p < 0.01.
Figure 4
Figure 4
Construction of macrophage-associated risk model and survival curve of high-/low-risk groups in TARGET-OS: (a) TARGET-OS was divided into high- and low-risk groups using the median risk score as the cutoff value, (b) the relationship between risk score and survival time and status of patients, (c) the survival curve of high- and low-risk groups in TARGET-OS, and (d) the heat map between expression of 5 MAGs and osteosarcoma samples.
Figure 5
Figure 5
Comparison of stromal and immune scores among high- and low-risk groups and survival analysis: (a) comparison of stromal score among high- and low-risk groups, (b) comparison of immune score among high- and low-risk groups, (c) survival analysis of different stromal scores among high- and low-risk groups (L: low, H: high, LSS: low stromal score, and HSS: high stromal score), and (d) survival analysis of different immune scores among high- and low-risk groups (L: low, H: high, LIS: low immune score, and HIS: high immune score).∗p < 0.05and ∗∗p < 0.01.
Figure 6
Figure 6
Identification of risk score as an independent prognostic factor of osteosarcoma, decision curve analysis of three models (simple gene model, simple clinical model, and complex model), and ROC curve for predicting the prognosis of osteosarcoma in 1, 3, and 5 years: (a) identification of risk score as an independent prognostic factor of osteosarcoma by univariate and multivariate Cox regression analyses, (b) the decision curve of the net benefit of the 2 models for the 3-year survival rate (simple gene and simple clinical models), (c) the decision curve of the net benefit of the 3 models for the 5-year survival rate (simple gene, simple clinical, and complex models); (d) the decision curve of the net benefit of the 3 models for the diagnosis of osteosarcoma metastasis (simple gene, simple clinical, and complex models), and (e) ROC curve to predict the prognosis of osteosarcoma in 1, 3, and 5 years.
Figure 7
Figure 7
Construction and internal validation of nomogram to predict the prognosis of osteosarcoma: (a) construction of the nomogram by collecting age, gender, tumor metastasis, race, and risk score; (b) 1-year survival calibration curve; (c) 3-year survival calibration curve; and (d) 5-year survival calibration curve.
Figure 8
Figure 8
Validation of macrophage-associated risk model and survival curve of high-/low-risk groups in GSE21257: (a) GSE21257 was divided into high- and low-risk groups using the median risk score as the cutoff value, (b) the relationship between risk score and survival time and status of patients, (c) the survival curve of high- and low-risk groups in GSE21257, and (d) ROC curve to predict the prognosis of osteosarcoma in 1, 3, and 5 years.
Figure 9
Figure 9
Validation of risk score as an independent prognostic factor of osteosarcoma in GSE21257 and important pathways identified by GSEA: (a) validation of risk score as an independent prognostic factor of osteosarcoma in GSE21257 by univariate and multivariate Cox regression analyses, (b) the important enrichment pathway: GO – macrophage migration, (c) the important enrichment pathway: GO-macrophage differentiation, and (d) the important enrichment pathways: GO-macrophage activation.
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References

    1. Kansara M., Teng M. W., Smyth M. J., Thomas D. M. Translational biology of osteosarcoma. Nature Reviews Cancer. 2014;14(11):722–735. doi: 10.1038/nrc3838. - DOI - PubMed
    1. Mirabello L., Troisi R. J., Savage S. A. Osteosarcoma incidence and survival rates from 1973 to 2004. Cancer. 2009;115(7):1531–1543. doi: 10.1002/cncr.24121. - DOI - PMC - PubMed
    1. Chou A. J., Kleinerman E. S., Krailo M. D., et al. Addition of muramyl tripeptide to chemotherapy for patients with newly diagnosed metastatic osteosarcoma. Cancer. 2009;115(22):5339–5348. doi: 10.1002/cncr.24566. - DOI - PMC - PubMed
    1. Link M. P., Goorin A. M., Miser A. W., et al. The effect OF adjuvant chemotherapy ON relapse-free survival IN patients with osteosarcoma OF the extremity. New England Journal of Medicine. 1986;314(25):1600–1606. doi: 10.1056/nejm198606193142502. - DOI - PubMed
    1. Hanahan D., Weinberg R. A. Hallmarks of cancer: the next generation. Cell. 2011;144(5):646–674. doi: 10.1016/j.cell.2011.02.013. - DOI - PubMed

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