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Radiosensitization by kinase inhibition revealed by phosphoproteomic analysis of pancreatic cancer cells.
Mol Cell Proteomics. 2020 Jul 10 [Online ahead of print]MC

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive cancers and known for its extensive genetic heterogeneity, high therapeutic resistance and strong variation in intrinsic radiosensitivity. In order to understand the molecular mechanisms underlying radioresistance, we screened the phenotypic response of 38 PDAC cell lines to ionizing radiation. Subsequent phosphoproteomic analysis of two representative sensitive and resistant lines led to the reproducible identification of 7,800 proteins and 13,000 phosphorylation sites (p-sites). Approximately 700 p-sites on 400 proteins showed abundance changes after radiation in all cell lines regardless of their phenotypic sensitivity. Apart from recapitulating known radiation response phosphorylation markers such as on proteins involved in DNA damage repair, the analysis uncovered many novel members of a radiation-responsive signaling network that was apparent only at the level of protein phosphorylation. These regulated p-sites were enriched in potential ATM substrates and in-vitro kinase assays corroborated 10 of these. Comparing the proteomes and phosphoproteomes of radiosensitive and -resistant cells pointed to additional tractable radioresistance mechanisms involving apoptotic proteins. For instance, elevated NADPH quinine oxidoreductase 1 (NQO1) expression in radioresistant cells may aid in clearing harmful reactive oxygen species. Resistant cells also showed elevated phosphorylation levels of proteins involved in cytoskeleton organization including actin dynamics and focal adhesion kinase (FAK) activity and one resistant cell line showed a strong migration phenotype. Pharmacological inhibition of the kinases FAK by Defactinib and of CHEK1 by Rabusertib showed a statistically significant sensitization to radiation in radioresistant PDAC cells. Together, the presented data map a comprehensive molecular network of radiation-induced signaling, improves the understanding of radioresistance and provides avenues for developing radiotherapeutic strategies.

Authors+Show Affiliations

Proteomics and Bioanalytics, Technical University of Munich, Germany.Department of Radiation Oncology, Technical University of Munich (TUM), Klinikum rechts der Isar, Germany.Department of Radiation Oncology, Technical University of Munich (TUM), Klinikum rechts der Isar, Germany.Technical University of Munich, Germany.Medical Clinic and Polyclinic II, University of Munich (TUM), Klinikum rechts der Isar, Germany.Medical Clinic and Polyclinic II, Technical University of Munich (TUM), Klinikum rechts der Isar, Germany.Department of Radiation Oncology, Technical University of Munich (TUM), Klinikum rechts der Isar, Germany.Department for Biosciences, Technical University of Munich, Germany.Department of Radiation Oncology, Technical University of Munich (TUM), Klinikum rechts der Isar, Germany sophie.dobiasch@tum.de.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32651227

Citation

Wiechmann, Svenja, et al. "Radiosensitization By Kinase Inhibition Revealed By Phosphoproteomic Analysis of Pancreatic Cancer Cells." Molecular & Cellular Proteomics : MCP, 2020.
Wiechmann S, Saupp E, Schilling D, et al. Radiosensitization by kinase inhibition revealed by phosphoproteomic analysis of pancreatic cancer cells. Mol Cell Proteomics. 2020.
Wiechmann, S., Saupp, E., Schilling, D., Heinzlmeir, S., Schneider, G., Schmid, R. M., Combs, S. E., Kuster, B., & Dobiasch, S. (2020). Radiosensitization by kinase inhibition revealed by phosphoproteomic analysis of pancreatic cancer cells. Molecular & Cellular Proteomics : MCP. https://doi.org/10.1074/mcp.RA120.002046
Wiechmann S, et al. Radiosensitization By Kinase Inhibition Revealed By Phosphoproteomic Analysis of Pancreatic Cancer Cells. Mol Cell Proteomics. 2020 Jul 10; PubMed PMID: 32651227.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Radiosensitization by kinase inhibition revealed by phosphoproteomic analysis of pancreatic cancer cells. AU - Wiechmann,Svenja, AU - Saupp,Elena, AU - Schilling,Daniela, AU - Heinzlmeir,Stephanie, AU - Schneider,Günter, AU - Schmid,Roland M, AU - Combs,Stephanie E, AU - Kuster,Bernhard, AU - Dobiasch,Sophie, Y1 - 2020/07/10/ PY - 2020/07/10/accepted PY - 2020/03/24/received PY - 2020/06/22/revised PY - 2020/7/12/entrez KW - Cancer therapeutics KW - Enzyme Inhibition* KW - Kinases* KW - Pancreatic cancer KW - Phosphoproteome KW - kinase inhibitors KW - kinase substrates KW - radioresistance JF - Molecular & cellular proteomics : MCP JO - Mol. Cell Proteomics N2 - Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive cancers and known for its extensive genetic heterogeneity, high therapeutic resistance and strong variation in intrinsic radiosensitivity. In order to understand the molecular mechanisms underlying radioresistance, we screened the phenotypic response of 38 PDAC cell lines to ionizing radiation. Subsequent phosphoproteomic analysis of two representative sensitive and resistant lines led to the reproducible identification of 7,800 proteins and 13,000 phosphorylation sites (p-sites). Approximately 700 p-sites on 400 proteins showed abundance changes after radiation in all cell lines regardless of their phenotypic sensitivity. Apart from recapitulating known radiation response phosphorylation markers such as on proteins involved in DNA damage repair, the analysis uncovered many novel members of a radiation-responsive signaling network that was apparent only at the level of protein phosphorylation. These regulated p-sites were enriched in potential ATM substrates and in-vitro kinase assays corroborated 10 of these. Comparing the proteomes and phosphoproteomes of radiosensitive and -resistant cells pointed to additional tractable radioresistance mechanisms involving apoptotic proteins. For instance, elevated NADPH quinine oxidoreductase 1 (NQO1) expression in radioresistant cells may aid in clearing harmful reactive oxygen species. Resistant cells also showed elevated phosphorylation levels of proteins involved in cytoskeleton organization including actin dynamics and focal adhesion kinase (FAK) activity and one resistant cell line showed a strong migration phenotype. Pharmacological inhibition of the kinases FAK by Defactinib and of CHEK1 by Rabusertib showed a statistically significant sensitization to radiation in radioresistant PDAC cells. Together, the presented data map a comprehensive molecular network of radiation-induced signaling, improves the understanding of radioresistance and provides avenues for developing radiotherapeutic strategies. SN - 1535-9484 UR - https://www.unboundmedicine.com/medline/citation/32651227/Radiosensitization_by_kinase_inhibition_revealed_by_phosphoproteomic_analysis_of_pancreatic_cancer_cells L2 - http://www.mcponline.org/cgi/pmidlookup?view=long&pmid=32651227 DB - PRIME DP - Unbound Medicine ER -
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