The most commonly reported primary malignant bone tumor in children and adolescents is osteogenic sarcoma (OGS) or osteosarcoma. Currently, neoadjuvant chemotherapy drugs, precision diagnostic instruments, limb salvage surgeries are the standard treatment procedures available for OGS patients. Recently, cryotherapy which uses extreme cold to destroy the cancer cells showing great results in bone cancer treatment. Thus, the usage of this treatment is widely increasing in OGS. The primary objective of this study is to identify the potential protein markers from the pre and post-cryotherapy treated bone samples of OGS patients (n=12) using liquid chromatography and tandem mass spectrometry with label-free quantification. This study successfully identified a common set of 23 significantly expressed proteins (p &lt;0.01) between two groups. Among these, 17 proteins were up-regulated with &gt;3 fold, and 6 proteins were down-regulated with &lt; 0.1 to 0.5 fold expression. The KEGG, GO and DAVID functional enrichment analysis revealed that the identified significant proteins are associated with cytoskeletal regulation, integrin, and VEGF signaling, TGF- beta, and p38 MAPK pathway, angiogenesis, glycolysis, and plasminogen activating cascades. Interestingly, the identified up-regulated proteins from OGS pre-cryotherapy samples showed dysregulated expressions in the post-cryotherapy treated OGS. Thus, we further validated these proteins using enzyme-linked immunosorbent assay (ELISA) with another set of OGS samples (n=10). The ELISA results revealed that the identified altered protein expression patterns among both the groups were consistently correlated in the verification analysis. Therefore, these protein candidates could serve as new biomarkers for bone malignancies, tumor microenvironment, apoptosis, and metastasis and also play an important role as therapeutic targets and sheds some light on precision medicine.

Identification of novel protein biomarkers using liquid chromatography/tandem mass spectrometry from pre and post-cryotherapy treated bones of osteosarcoma patients

2019 Spring National Meeting

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