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. 2025 Jul 19:24:11769351251331508.
doi: 10.1177/11769351251331508. eCollection 2025.

Single-Cell Transcriptome Analyses of Four Pain Related Genes in Osteosarcoma

Mesalie Feleke  1 , Haiyingjie Lin  1 , Yun Liu  2 , Liang Mo  3 , Emel Rothzerg  1 , Dezhi Song  2 , Jinmin Zhao  2 , Wenyu Feng  4 , Jiake Xu  1   3
Affiliations

Single-Cell Transcriptome Analyses of Four Pain Related Genes in Osteosarcoma

Mesalie Feleke et al. Cancer Inform. .

Abstract

Objective: Osteosarcoma (OS) is a rare and complex form of cancer that mostly affects children and adolescents. Pain is a common symptom for patients in OS which causes significant unhappiness and persistent aches. To date, there is minimal knowledge on the mechanisms underlying OS induced pain and few treatment options for patients. Previous genetic studies have demonstrated that the panel of four genes, artemin (ARTN), persephin (PSPN), glial cell line-derived neurotropic factor (GDNF), and neurturin (NRTN) are associated with the regulation of pain processing in OS and analgesic responses.

Methods: In the present study, by utilising a scRNA-seq OS dataset, we aimed to measure the gene expression levels of four pain related genes, and compare them between the different cell types in human OS tissues and cell lines.

Results: Within a complex and diverse range of cell types in OS tissues, including osteoblastic OS cells, carcinoma associated fibroblasts (CAFs), B cells, myeloid cells 1, myeloid cells 2, NK/T cells, plasmocytes, ARTN and NRTN genes had the highest expression in Osteoblastic OS cells, GDNF gene had a peak expression in carcinoma associated fibroblasts, and PSPN gene in endothelial cells. In addition, all four genes showed deferential pattern of expression in 16 OS cell lines.

Conclusion: Future studies should investigate the potential to target deferentially expressed pain-related genes in specific cell types of OS for therapeutic benefit to improve the quality of life for patients living with pain caused by OS.

Keywords: NGS; bioinformatics; osteoblastic cells; osteosarcoma; sarcoma; scRNA sequencing.

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Conflict of interest statement

The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Figures

Figure 1.
Figure 1.
Violin plots showing the gene expression levels of ARTN (A), GDNF (B), NRTN (C), and PSPN (D) in OS and normal tissues. The violin plots filling in red representing normal groups, and the blue plots represent OS groups. The y-axis indicates the expression level of the genes. By using Student’s t-test, P < .05 was considered statistically significant (***P < .001, ****P < .00001).
Figure 2.
Figure 2.
Uniform Manifold Approximation and Projection (UMAP) depiction of scRNA-seq data showing the 9 main cell types in OS.
Figure 3.
Figure 3.
Uniform Manifold Approximation and Projection (UMAP) plot showing the relative mRNA expression in TPM of ARTN (A), GDNF (B), NRTN (C), and PSPN (D) in the 9 main OS cell types.
Figure 4.
Figure 4.
Dot plot displaying expression levels in TPM of four pain related genes in diverse OS cells.
Figure 5.
Figure 5.
Heatmap displaying mRNA gene expression in TPM of four pain related genes PSPN, NRTN, ARTN, and GDNF in 9 OS cell types.
Figure 6.
Figure 6.
Genes expression in Cancer Cell Line Encyclopedia (CCLE) datasets. Bar plot displaying mRNA gene expression in TPM of four pain related genes (PSPN, NRTN, ARTN, and GDNF) in 16 OS cell lines.
Figure 7.
Figure 7.
Plots of Kaplan-Meier analysis of the survival rate of OS patients with the expression levels of ARTN (A), GDNF (B), NRTN (C), and PSPN (D) genes. Red colour represents genes with high expression, whereas blue colour represents genes with low expression. No significant difference in survival probability was found between high and low expression of these genes in OS patients.
See this image and copyright information in PMC

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