Characterization of Novel Biomarkers for Prostate Cancer Diagnosis Using Mass Spectrometry-based Proteomics

Veronica Vicente^*
*Correspondence: Veronica Vicente, Department of Medical and Clinical Biophysics, University of Pavol Jozef Šafárik in Košice, Trieda SNP 1, 04011 Košice, Slovakia, Email:

Keywords

Prostate cancer • Biomarkers • Mass spectrometry

Introduction

Prostate cancer is the second most common cancer in men worldwide, with early detection being crucial for effective treatment and improved patient outcomes. Current diagnostic methods, including Prostate-Specific Antigen (PSA) testing, suffer from limitations in sensitivity and specificity, often leading to overdiagnosis and unnecessary biopsies. Therefore, there is an urgent need for more reliable biomarkers that can accurately distinguish prostate cancer from benign conditions and other malignancies. Advances in mass spectrometry-based proteomics offer a powerful approach for the comprehensive analysis of protein expression profiles, enabling the discovery of novel biomarkers with potential clinical utility [1].

Literature Review

This study employed a comprehensive mass spectrometry-based proteomics approach to identify novel biomarkers for the diagnosis of prostate cancer. The workflow involved several key steps, from sample collection and preparation to data analysis and biomarker validation. Serum samples were obtained from two well-defined cohorts: patients diagnosed with prostate cancer and age-matched healthy controls. All participants provided informed consent, and the study protocol was approved by the relevant institutional review board. Blood samples were collected in the morning after fasting and processed to obtain serum, which was aliquoted and stored at -80°C until analysis [2].

Proteins were extracted from serum samples using a precipitation method to remove high-abundance proteins and enrich low-abundance proteins, which are often more relevant for disease biomarker discovery. The extracted proteins were then quantified using a Bicin Choninic Acid (BCA) assay to ensure consistent protein input for subsequent analysis. The protein samples were subjected to enzymatic digestion using trypsin, resulting in a complex mixture of peptides. To enable multiplexed quantitative analysis, the digested peptides were labeled using isobaric tags for relative and absolute quantitation (iTRAQ). This labeling technique allows for the simultaneous comparison of multiple samples within a single mass spectrometry run, increasing throughput and reducing experimental variability.

The iTRAQ-labeled peptides were fractionated by High-Performance Liquid Chromatography (HPLC) to reduce sample complexity before being introduced into the mass spectrometer. The peptides were then analyzed using a high-resolution tandem mass spectrometer, which provided detailed information on peptide sequences and their relative abundances across different samples. The raw mass spectrometry data were processed using specialized bioinformatics software. This involved database searching to match the acquired spectra to known protein sequences, followed by quantification of peptide and protein abundances. Differential expression analysis was conducted to identify proteins that showed significant differences in expression levels between prostate cancer patients and healthy controls [3].

A list of candidate biomarkers was generated based on statistical criteria, including fold change and p-value thresholds. Proteins that were significantly upregulated or downregulated in prostate cancer patients were prioritized for further validation. To validate the initial findings, a targeted proteomics approach known as multiple reaction monitoring (MRM) was employed. MRM allows for the precise and accurate quantification of selected peptides in complex biological samples. This validation step involved synthesizing stable isotope-labeled standard peptides for the candidate biomarkers and using them as internal standards in the MRM assays [4].

Discussion

The mass spectrometry-based proteomics approach enabled the comprehensive profiling of serum proteins, leading to the identification of several novel biomarkers associated with prostate cancer. Among the differentially expressed proteins, several were found to be upregulated or downregulated significantly in prostate cancer patients compared to healthy controls. These proteins are involved in various biological processes relevant to cancer pathogenesis, including cell proliferation, apoptosis, and immune response. The validation of these biomarkers using targeted proteomics confirmed their diagnostic potential, with some candidates demonstrating high sensitivity and specificity in distinguishing prostate cancer from non-cancerous conditions. The ROC curve analysis further underscored the diagnostic accuracy of these biomarkers, suggesting their utility in clinical settings [5]. Importantly, the discovery of these novel biomarkers offers new insights into the molecular mechanisms underlying prostate cancer, potentially guiding the development of targeted therapies and personalized treatment strategies. However, further validation in larger, independent cohorts is necessary to confirm the clinical applicability of these findings [6].

Conclusion

This study highlights the power of mass spectrometry-based proteomics in identifying and characterizing novel biomarkers for prostate cancer diagnosis. The identified biomarkers exhibit promising diagnostic potential, offering a significant improvement over current PSA testing. Continued research and validation efforts are essential to translate these findings into clinical practice, ultimately enhancing early detection and treatment outcomes for prostate cancer patients. The integration of these biomarkers into diagnostic protocols could revolutionize prostate cancer management, reducing overdiagnosis and enabling more precise, individualized care.

Acknowledgement

None.

Conflict of Interest

None.

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