Proteiny odpovědné za špatnou terapeutickou odpověď metastatických nádorů ledvin na léčbu tyrosinkinasovými inhibitory
| Title in English | Proteins associated with low response to tyrosinkinase inhibitor therapy in metastatic kidney tumors |
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| Authors | |
| Year of publication | 2022 |
| Type | Conference abstract |
| MU Faculty or unit | |
| Citation | |
| Description | Metastatic renal carcinoma (mRCC) is cancer with poor prognosis. The patients are treated with tyrosinkinase inhibitors (TKI), but many of patients response insufficiently to this therapy. Up to present, there is no molecular marker identifying patients, who will response poorly and who would benefit from alternative type of therapy (immunotherapy, experimental clinic studies). Aim of the study is to apply proteomics methods for identification of proteins or proteins panels, which could serve as predictive markers of poor response to TKI therapy. The proteomics analysis of 75 mRCC tissues from patients treated with TKI sunitinib or pazopanib in first line, from whose 42 patients were non-responders, and of 17 normal kidney tissues was done using mass spectrometry with data independent analysis (DIA-MS) on QExactive HF-X mass spectrometer (Thermo Fisher Scientific). 6183 protein groups were quantified (Qvalue<0.01). The classificator with predictive potential, which includes 10 selected proteins, was constructed based on the proteomic data. Transmembrane glycoprotein GPNMB showed increased protein level in TKI non-responsive tumors in comparison with responsible ones and could serve as a potential diagnostic and therapeutic target. GPNMB gene was disrupted in 786-0 cell line derived from renal carcinoma using CRISPR/Cas 9 method and a 10 bp deletion with frameshift leading to stop codon was identified, decreased protein level was confirmed in immunochemical detection of GPNMB using western blot. The “scratch assay” was performed to reveal changes of migratory properties in such modified 786-0 cells. Cell with decreased level of GPNMB migrated slowly in comparison to parental cells. The methods for routinely assessment of GPNMB protein were established: a targeted single reaction monitoring (SRM) MS method and immunohistochemical staining of tissue slices. In conclusion, proteomics analysis based on DIA-MS enables identification of new potential diagnostic and therapeutic targets in mRCC. Increased protein level of GPNMB is associated with poor response of mRCC patients to TKI therapy and supports migrative capabilities of cancer cells. Supported by Ministry of Health of the Czech Republic, project No. NV19-08-00250, all rights reserved, and by The project National Institute for Cancer Research (Programme EXCELES, ID Project No. LX22NPO5102) - Funded by the European Union - Next Generation EU. CIISB, Instruct-CZ Centre of Instruct-ERIC EU consortium, funded by MEYS CR infrastructure project LM2018127, is gratefully acknowledged for the financial support of the measurements at the CEITEC Proteomics Core Facility. |
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