ATG16L1 is a Potential Prognostic Biomarker and Immune Signature for Osteosarcoma: A Study Based on Bulk RNA and Single-Cell RNA-Sequencing

Abstract

Zhaojie Qin,1,2,* Kai Luo,1,* Yun Liu,1 Shijie Liao,3 Juliang He,4 Mingwei He,3 Tianyu Xie,3 Xiaohong Jiang,5 Boxiang Li,5 Huijiang Liu,1,6 Qian Huang,6 Haijun Tang,5 Wenyu Feng,3,* Xinli Zhan1,* 1Department of Spinal Bone Disease, First Affiliated Hospital of Guangxi Medical University, Nanning, 530021, Guangxi, People’s Republic of China; 2Department of Orthopedic, The People’s Hospital of Hechi, Hechi, 547600, Guangxi, People’s Republic of China; 3Department of Trauma Orthopedic and Hand Surgery, First Affiliated Hospital of Guangxi Medical University, Nanning, 530021, Guangxi, People’s Republic of China; 4Department of Bone and Soft Tissue Surgery, Guangxi Medical University Cancer Hospital, Nanning, 530021, Guangxi, People’s Republic of China; 5Department of Orthopedic, Affiliated Minzu Hospital of Guangxi Medical University, Nanning, 530021, Guangxi, People’s Republic of China; 6Department of Orthopedics, The First People’s Hospital of Nanning, Nanning, 530021, Guangxi, People’s Republic of China*These authors contributed equally to this workCorrespondence: Xinli ZhanDepartment of Spinal Bone Disease, First Affiliated Hospital of Guangxi Medical University, Nanning, 530021, Guangxi, People’s Republic of China, Tel [phone], Fax [phone], Email [email protected]; Wenyu FengDepartment of Trauma Orthopedic and Hand Surgery, First Affiliated Hospital of Guangxi Medical University, Nanning, 530021, Guangxi, People’s Republic of China, Tel [phone], Fax [phone], Email [email protected]: Osteosarcoma is a common solid malignancy of the bone in children and adolescents, and its metastasis and recurrence are the principal causes of poor treatment outcomes.Methods: Autophagy-related genes were used to cluster osteosarcoma patients by consensus clustering analysis using the GSE21257 database. Differentially expressed genes (DEGs) were identified by limma package. Multiple-gene risk signature was constructed using least absolute shrinkage and selection operator (LASSO) analysis and Cox regression analyses. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to determine gene expression levels. Then, single-cell RNA-sequencing dataset GSE152048 were used to identify the correlation between the DEGs and effector molecules expressed in specific tumor-infiltrating immune cells.Results: Two clusters were identified in the consensus clustering analysis, which were confirmed by principal component analysis. Limma analysis revealed that 15 genes were related, and 9 genes were screened using protein-protein interaction network and LASSO regression analysis. Cox regression analyses identified 5 genes. Combined with survival analysis, only the autophagy related 16 like 1 gene (ATG16L1) was significant. The results of qRT-PCR showed low expression levels of ATG16L1 in tumor cells group. Immune infiltration analysis revealed significantly lower expression of CD8+ T cells in the high ATG16L1 gene expression group. ScRNA-seq revealed that in the ATG16L1+CD8+ T cell group, the expression of GZMB was lower, whereas the expression of ITGA1 was higher. These results showed that ATG16L 1 is an immune-related gene, which is associated with poor prognosis in patients with osteosarcoma.Conclusion: ATG16L1 is a potential prognostic biomarker and immune signature and may be a therapeutic target for osteosarcoma.Keywords: osteosarcoma, prognosis, ATG16L1, CD8+ T cell, scRNA-seq

Keywords

• osteosarcoma
• prognosis
• atg16l1
• cd8+ t cell
• scrna-seq