LAPČÍK, Petr, Lucia JANÁČOVÁ, Pavla BOUCHALOVÁ, David POTĚŠIL, Ján PODHOREC, Milan HORA, Alexandr POPRACH, Ondřej FIALA and Pavel BOUCHAL. A large-scale assay library for targeted protein quantification in renal cell carcinoma tissues. In 20th Human Proteome Organization World Congress, HUPO Reconnect 2021, Online, 15.11.-19.11.2021, in Book of Abstracts, p. 184. 2021.
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Basic information
Original name A large-scale assay library for targeted protein quantification in renal cell carcinoma tissues
Name in Czech Rozsáhlá knihovna cílených hmotnostně-spektrometrických assays pro kvantifikaci proteinů v tkáních renálního karcinomu
Authors LAPČÍK, Petr, Lucia JANÁČOVÁ, Pavla BOUCHALOVÁ, David POTĚŠIL, Ján PODHOREC, Milan HORA, Alexandr POPRACH, Ondřej FIALA and Pavel BOUCHAL.
Edition 20th Human Proteome Organization World Congress, HUPO Reconnect 2021, Online, 15.11.-19.11.2021, in Book of Abstracts, p. 184. 2021.
Other information
Original 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
WWW URL
Organization unit Faculty of Science
Keywords in English assay library, data independent acquisition, mass spectrometry, proteomics, renal cell carcinoma
Tags International impact
Changed by Changed by: Mgr. Petr Lapčík, Ph.D., učo 409180. Changed: 9/2/2022 16:37.
Abstract
Introduction. Renal cell carcinoma (RCC) represents 2.2 % of cancer incidences, however, prognostic or predictive protein biomarkers specific for RCC are generally not available. To support proteomics research of localized and metastatic RCC, we introduce a new library of targeted mass spectrometry assays for accurate protein quantification in malignant and normal kidney tissue. Methods. Aliquots of 86 initially localized RCC, 75 metastatic RCC and 17 adjacent non-cancerous fresh frozen tissue lysates were trypsin digested, pooled and fractionated using hydrophilic chromatography and analyzed using LC-MS/MS on QExactive HF-X mass spectrometer in data-dependent acquisition mode. The library was generated in Spectronaut software. Two published datasets A-B [1] [2] and two new pilot datasets C-D of localized and metastatic RCC tissues measured in data-independent acquisition (DIA) mode were processed using the new library in Spectronaut. Results. The newly established assay library contains 77,817 peptides representing 7960 protein groups (FDR=1%). Its application resulted in increased numbers of quantified proteins in datasets A (2463 proteins, +4%) and B (4492 protein groups, >2 fold), with a clear separation of tumor and non-tumor tissues in both studies. Analysis of datasets C of metastatic RCC responding vs. non-responding to sunitinib treatment and dataset D of initially localized vs. metastatic RCC tissues led to consistent quantification of 5181 and 5253 protein groups (FDR=1%). Conclusions. Application of our spectral library leads to quantification of substantially increased part of RCC proteome. The new library has potential to contribute to better understanding the RCC development at molecular level, leading to new diagnostic and therapeutic targets.
Links
LM2018127, large research infrastructuresName: Czech Infrastructure for Integrative Structural Biology (Acronym: CIISB)
Investor: Ministry of Education, Youth and Sports of the CR
LM2018127, large research infrastructuresName: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)
Investor: Ministry of Education, Youth and Sports of the CR
MUNI/A/1604/2020, interní kód MUName: Podpora biochemického výzkumu v roce 2021
Investor: Masaryk University
NV19-08-00250, research and development projectName: Proteotypová klasifikace renálního karcinomu ve vztahu k prognóze a terapeutické odpovědi
Investor: Ministry of Health of the CR
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