DIA-MS Identifies and Validates Transgelin as Protein Contributing to a Poor Response of Metastatic Renal Cell Carcinoma to Sunitinib Treatment
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| Year of publication | 2021 |
| Type | Conference abstract |
| MU Faculty or unit | |
| Citation | |
| Description | Introduction. Renal cell carcinoma (RCC) represents about 2–3% of all cancers with over 400,000 new cases per year. Sunitinib, a vascular endothelial growth factor tyrosine kinase receptor inhibitor, has been used mainly for first-line treatment of metastatic clear-cell RCC (mccRCC) with good or intermediate prognosis. However, about one-third of mccRCC patients do not respond to sunitinib, leading to disease progression. Methods. Here, we aim to find and characterize proteins associated with poor sunitinib response. Sixteen RCC tumors from patients responding (8) vs. non-responding (8) to sunitinib 3 months after treatment initiation were analyzed using data-independent acquisition mass spectrometry (DIA-MS) on Impact II LC-MS system (Bruker), together with their adjacent non-cancerous tissues in a pilot proteomics study. Validation was performed in an independent cohort of 75 mccRCC patients using DIA-MS on QExactive HF-X (Thermo Fisher Scientific), with data analysis in Spectronaut 13.9 (Biognosys). Transgelin protein role was functionally analyzed in RCC cell lines using CRISPR and RNAi silencing techniques. Results. Proteomics analysis quantified 1996 protein groups (FDR=0.01) and revealed 27 proteins deregulated between tumors non-responding vs. responding to sunitinib in the pilot study, representing a pattern of proteins potentially contributing to sunitinib resistance. Validation cohort confirmed up-regulation of transgelin in tumors non-responding to sunitinib and revealed its association with tumor grade. Gene set enrichment analysis showed an up-regulation of epithelial-to-mesenchymal transition with transgelin as one of the most significantly abundant proteins. Reduced transgelin protein level in CRISPR and RNAi altered RCC cells led to significantly slower proliferation of these cells and affected their survival. Conclusion. Altogether, our data indicate that transgelin is an essential protein supporting RCC cell proliferation, which could contribute to intrinsic sunitinib resistance, and is associated with aggressive phenotype of RCC tumors. |
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