Quantification of cancer biomarkers using targeted proteomic approach

• Grant number: 13/21603-5
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate (Direct)
• Start date: March 01, 2014
• End date: May 31, 2014
• Field of knowledge: Biological Sciences - Biochemistry
• Principal Investigator: Adriana Franco Paes Leme
• Grantee: Rebeca Kawahara Sakuma
• Supervisor: Michael J. MacCoss
• Host Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia e Inovação (Brasil). Campinas , SP, Brazil
• Institution abroad: University of Washington, United States
• Associated to the scholarship: 11/22421-2 - Determination of cleavage sites of recombinant ADAM-17 in human cells, BP.DD

Abstract

Verification of candidate biomarkers relies upon specific, quantitative assays optimized for selective detection of target proteins. Despite the large numbers of putative biomarkers, only a small number of them are passed through the development and validation process into clinical practice, and their rate of introduction is declining. Selected reaction monitoring (SRM) is a targeted mass spectrometry approach that has been emerging as a promising technique to precisely and quantitatively analyze complex biological samples. Due to its high specificity, sensitivity, multiplexing capability, and precision, SRM provides a powerful method for candidate biomarker verification. In our group, we started studying cancer biomarkers by analyzing the secretome composition of different cancer cell lines and oral squamous cell carcinoma (OSCC) microdissected from oral cancer tissue biopsies. In this step, we have raised many candidates biomarkers based on discovery proteomics experiments, which in a second step, need targeted proteomics to be validated. Thus, this project aims to develop SRM-MS experiments using isotopically labeled reference peptides for quantifying, in complex matrices, the candidate markers found in our studies. The collaborative training with expertise in this field will be extremely important to prepare people to design, analyze and interpret targeted proteomics. We also need expertizes to implement this technology in Brazil, which will impact not only the quality and relevance of our researches but also the importance of our laboratory as a mass spectrometry facility open to share infrastructure, cutting-edge research and innovation to the academic and industrial Brazilian community. (AU)