Proteomic Identification of Biomarkers in the Cerebrospinal Fluid (CSF) of Astrocytoma Patients

Abstract

The monitoring of changes in the protein composition of the cerebrospinal fluid (CSF) can be used as a sensitive indicator of central nervous system (CNS) pathology, yet its systematic application to analysis of CNS neoplasia has been limited. There is a pressing need for both a better understanding of gliomagenesis, and the development of reliable biomarkers of the disease. In this report, we used two proteomic techniques, two-dimensional gel electrophoresis (2-DE) and cleavable Isotope-Coded Affinity Tag (cICAT), to compare CSF proteomes in order to identify tumor and grade specific biomarkers in patients bearing brain tumors of differing histologies and grades. Retrospective analyses were performed on 60 samples derived from astrocytomas WHO grade II, III and IV, schwannomas, metastastic brain tumors, inflammatory samples and non-neoplastic controls. We identified 103 potential tumor-specific markers; of which 20 were high-grade astrocytoma-specific. These investigations allowed us to identify a spectrum of signature proteins that could differentiate *Address for Correspondence/Reprints: Erwin G. Van Meir, Ph.D, Winship Cancer Institute, Emory University, 1365C Clifton Rd. N.E, C5078, Atlanta, GA 30322, Phone: 404-778-5563, Fax: 404-778-5550, e-mail: evanmei@emory.edu. Significance: The measurement of alterations in the protein composition of the cerebrospinal fluid (CSF) can be used as a sensitive indicator of central nervous system (CNS) pathology, yet its systematic application to analysis of CNS neoplasia has been limited. There is a pressing need for both a better understanding of gliomagenesis, and the development of reliable biomarkers of the disease. Our investigations allowed us to identify a spectrum of signature proteins that could differentiate between low (AII) and high-grade (AIV) astrocytoma that warrant further validation as potential new markers for diagnosis, prognosis and disease follow-up . These candidate biomarkers may also have functional properties that play a critical role in the development and malignant progression of human astrocytomas, thus possibly representing novel therapeutic targets for this highly lethal disease.Author contributions Project concept and experimental designs were developed by EGVM and FWK. All experiments were perfomed by FWK with the following exceptions; Mass spectrometry and cICAT were done by MSR. BJS performed the statistical analysis, JP provided expertise with proteomic analyses; JJO, MDG, AG, SK and GYG all provided CSF samples. FWK and EGVM interpreted the results and wrote the manuscript. All authors read the manuscript. NIH Public Access

Detection of tumor-derived DNA in cerebrospinal fluid of patients with primary tumors of the brain and spinal cord

“…Future studies will be required to directly compare CSF-tDNA with CSF cytology. Rather than replacing cytology, we envision that CSF-tDNA will be used in combination with it and other biomarkers under development as well as with radiographic and clinical parameters (35)(36)(37)(38). This could substantially increase the accuracy of the estimates of tumor burden at various points during the management of patients.…”

Biomarkers of Glioma

“…These techniques include ELISA (Hormigo et al, 2006;Lin et al, 2009;Sreekanthreddy et al, 2010), 2-DE (Khwaja et al, 2007;Khalil, 2007;Ohnishi et al, 2009), DNA microarray , (Petrik et al, 2008;Schwartz et al, 2005),direct sequencing (Sanson et al, 2009), ICAT (Khwaja et al, 2007), and transgenic mouse etc (Sodaa et al, 2011). A number of quantitative approaches summarized in Figure 1, with different technical advantages and challenges, have been developed for glioma biomarker discovery.…”

“…Furthermore, CSF has about 1000 proteins, which is 100~400 fold lower than in serum, and this will be facilitated to screen the specific protein biomarkers (Omenn, 2005). In the past decades, many markers have been found in CSF of glioma patients, such as S100, neuronspecific enolase (NSE), recoverin (protein A) and vascular endothelial growth factor (VEGF) (Gronowitz et al, 1984;Cochran & Wen, 1985;Taomoto et al, 1987;Sampath et al, 2004;Khwaja et al, 2007). However, these biomarkers could not be widespread used due to the limitation of repetition in CSF examinations.…”

“…Recently, new candidate biomarkers have been identified by proteomics analysis for CSF samples. For example, Khwaja et al used two-dimensional gel electrophoresis (2-DE) and cleavable Isotope-Coded Affinity Tag (cICAT) to analyze CSF proteome to detect specific biomarkers in brain tumors with differing histologies and grades (Khwaja et al, 2007). Finally, 20 potential tumor-specific markers were specially identified in high-grade astrocytoma.…”

Preliminary use of differential scanning calorimetry of cerebrospinal fluid for the diagnosis of glioblastoma multiforme

“…The normal CSF protein concentration is 100-to 400-fold lower than in serum, allowing a much greater signal-to-noise ratio for detecting biomarker proteins [1][2][3][4]. As proof of principle, human CSF has been examined using proteomic approaches, and proteins that might be used as markers of tumor aggressiveness and invasiveness have been identified [5]. Although this approach requires sophisticated molecular techniques and is labor and time intensive, it suggests that the use of CSF protein profiles might allow for reliable methods of determining tumor characteristics directly without invasive intracranial procedures.…”