DIA-MS Identifies and Validates Transgelin as Protein
Contributing to a Poor Response of Metastatic Renal Cell
Carcinoma to Sunitinib Treatment

a 2021

DIA-MS Identifies and Validates Transgelin as Protein Contributing to a Poor Response of Metastatic Renal Cell Carcinoma to Sunitinib Treatment

BOUCHALOVÁ, Pavla; Jindřich BERÁNEK; Petr LAPČÍK; David POTĚŠIL; Ján PODHOREC et. al.

Basic information

Original name

DIA-MS Identifies and Validates Transgelin as Protein Contributing to a Poor Response of Metastatic Renal Cell Carcinoma to Sunitinib Treatment

Name in Czech

DIA-MS analýza identifikovala a validovala transgelin jako protein přispívající ke slabé odpovědi na léčbu sunitinibem u metastatického karcinomu ledvin

Authors

BOUCHALOVÁ, Pavla; Jindřich BERÁNEK; Petr LAPČÍK; David POTĚŠIL; Ján PODHOREC; Milan HORA; Ondřej FIALA; Alexandr POPRACH and Pavel BOUCHAL

Edition

HUPO Reconnect 2021, 2021

Other information

Language

English

Type of outcome

Conference abstract

Field of Study

10608 Biochemistry and molecular biology

Country of publisher

United States of America

Confidentiality degree

is not subject to a state or trade secret

References:

URL

Organization unit

Faculty of Science

Keywords in English

renal carcinoma, sunitinib, resistance, DIA proteomics, transgelin, silencing technique, proliferation, survival

Abstract

In the original language

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.

Links

MUNI/A/1604/2020, interní kód MU

Name: Podpora biochemického výzkumu v roce 2021

Investor: Masaryk University

NV19-08-00250, research and development project

Name: Proteotypová klasifikace renálního karcinomu ve vztahu k prognóze a terapeutické odpovědi

Investor: Ministry of Health of the CR

Cite

BOUCHALOVÁ, Pavla; Jindřich BERÁNEK; Petr LAPČÍK; David POTĚŠIL; Ján PODHOREC; Milan HORA; Ondřej FIALA; Alexandr POPRACH and Pavel BOUCHAL. DIA-MS Identifies and Validates Transgelin as Protein Contributing to a Poor Response of Metastatic Renal Cell Carcinoma to Sunitinib Treatment. In HUPO Reconnect 2021. 2021.

@proceedings{1804898,
   author = {Bouchalová, Pavla and Beránek, Jindřich and Lapčík, Petr and Potěšil, David and Podhorec, Ján and Hora, Milan and Fiala, Ondřej and Poprach, Alexandr and Bouchal, Pavel},
   booktitle = {HUPO Reconnect 2021},
   keywords = {renal carcinoma, sunitinib, resistance, DIA proteomics, transgelin, silencing technique, proliferation, survival},
   language = {eng},
   title = {DIA-MS Identifies and Validates Transgelin as Protein Contributing to a Poor Response of Metastatic Renal Cell Carcinoma to Sunitinib Treatment},
   url = {https://hupo2021.org/wp-content/uploads/2021/11/HUPO-2021-Abstract-Book_Oct-27.pdf},
   year = {2021}
}

TY  - CONF
ID  - 1804898
AU  - Bouchalová, Pavla - Beránek, Jindřich - Lapčík, Petr - Potěšil, David - Podhorec, Ján - Hora, Milan - Fiala, Ondřej - Poprach, Alexandr - Bouchal, Pavel
PY  - 2021
TI  - DIA-MS Identifies and Validates Transgelin as Protein Contributing to a Poor Response of Metastatic Renal Cell Carcinoma to Sunitinib Treatment
KW  - renal carcinoma, sunitinib, resistance, DIA proteomics, transgelin, silencing technique, proliferation, survival
UR  - https://hupo2021.org/wp-content/uploads/2021/11/HUPO-2021-Abstract-Book_Oct-27.pdf
N2  - 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.
ER  -

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