Short Communication Multiplexed Immunobead-Based Cytokine Profiling for Early Detection of Ovarian Cancer Elieser Gorelik, 1,5 Douglas P. Landsittel, 2 Adele M. Marrangoni, 5 Francesmary Modugno, 3,5 Lyudmila Velikokhatnaya, 5 Matthew T. Winans, 5 William L. Bigbee, 5 Ronald B. Herberman, 4,5 and Anna E. Lokshin 4,5 Departments of 1 Pathology and Immunology, 2 Biostatistics, and 3 Epidemiology, 4 Division of Hematology/Oncology, 5 University of Pittsburgh School of Medicine and University of Pittsburgh Cancer Institute, Pittsburgh, Pennsylvania Abstract Early detection of ovarian cancer might improve clinical outcome. Some studies have shown the role of cytokines as a new group of tumor markers for ovarian cancer. We hypothesized that a panel comprised of multiple cytokines, which individually may not show strong correlation with the disease, might provide higher diagnostic power. To evaluate the diagnostic utility of cytokine panel, we used a novel multianalyte LabMAP profiling technology that allows simultaneous measurement of multiple markers. Concen- trations of 24 cytokines (cytokines/chemokines, growth, and angiogenic factors) in combination with cancer antigen-125 (CA-125), were measured in sera of 44 patients with early- stage ovarian cancer, 45 healthy women, and 37 patients with benign pelvic tumors. Six markers, i.e., interleukin (IL)-6, IL- 8, epidermal growth factor (EGF), vascular endothelial growth factor (VEGF), monocyte chemoattractant protein-1 (MCP-1), and CA-125, showed significant differences in serum concentrations between ovarian cancer and control groups. Out of this group, IL-6, IL-8, VEGF, EGF, and CA- 125, were used in a classification tree analysis that resulted in 84% sensitivity at 95% specificity. The receiver operator characteristic curve created using the combination of markers produced sensitivities between 90% and 100% in the area of 80% to 90% specificity, whereas the receiver operator characteristic curve for CA-125 alone resulted in sensitivities of 70% to 80%. The classification tree analysis for discrimination of benign condition from ovarian cancer used CA-125, granulocyte colony-stimulating factor (G-CSF), IL-6, EGF, and VEGF resulting in 86.5% sensitivity and 93.0% specificity. The presented data show that simulta- neous testing of a panel of serum cytokines and CA-125 using LabMAP technology may present a promising ap- proach for ovarian cancer detection. (Cancer Epidemiol Biomarkers Prev 2005;14(4):981 – 7) Introduction Ovarian cancer represents the third most frequent cancer of the female genital tract. The majority of early-stage ovarian cancers are asymptomatic, and over three-quarters of clinical diagnoses are made at a time when the disease has already established regional or distant metastases. Despite aggressive cytoreductive surgery and platinum-based chemotherapy, the 5-year survival for patients with clinically advanced ovarian cancer is only 15% to 20%, in striking contrast to the cure rate for stage I disease, which is usually >90% (1). These statistics provide the primary rationale to improve ovarian cancer screening and early detection. Due to the low prevalence of spontaneous ovarian cancer in the general population, a screening strategy must have sensitivity of at least 80% in early-stage disease and near- perfect specificity of at least 99.6% (2). At present, there are two screening tests for ovarian cancer: serologic screening for tumor antigen using cancer antigen-125 (CA-125), and imaging using transvaginal sonography (2-6). However, with a cutoff of 30 to 35 units/mL, serum CA-125 has been shown to have a sensitivity of only 50% to 60% with the specificity of >98%, for early-stage disease (4, 7, 8). Transvaginal sonography alone or combined with Doppler and morphologic indices, are only sensitive and specific for established tumors, and are, therefore, not suitable for early diagnostics of ovarian cancer (6, 9). Recently, a novel technology, surface-enhanced laser desorption/ionization time-of-flight mass spectrometry has been offered for early detection of ovarian cancer (10). This technology was reported to allow for discriminating serum protein profiles with 100% sensitivity and 100% specificity (11). However, in two other studies of early detection of ovarian cancer using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry, the results were less opti- mistic, demonstrating 72.8% to 95.7% sensitivity and 82.6% to 94.9% specificity (12, 13). Therefore, at present, proteomic profiling, whereas promising, does not possess the required diagnostic discrimination for primary ovarian cancer screen- ing. Additional approaches are necessary to provide the required high level of specificity and positivity for an effective high-throughput screening for ovarian cancer. During the last two decades, a large number of serologic tumor markers have been evaluated for their ability to detect early-stage epithelial ovarian cancer. Biomarkers that have a shown association with ovarian cancer include cancer antigens, differentiation markers, antibodies to mutated onco- genes, and cytokines (reviewed in ref. 14). Cytokines are a diverse group of proteins comprised of hematopoietic growth factors, interferons, lymphokines, and chemokines (15). Serum cytokines that possess diagnostic value in ovarian cancer include interleukin (IL)-6, IL-8, macrophage colony-stimulating Received 6/3/04; revised 11/22/04; accepted 12/10/04. Grant support: NIH grant 1RO1 CA098642-01A1 and the DOD grant DAMD17-03-1-0696 (A.E. Lokshin), and NIH grant RO3 CA102888 (E. Gorelik). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. Requests for reprints: Anna E. Lokshin, Hillman Cancer Center, University of Pittsburgh Cancer Institute, 5117 Centre Avenue, Pittsburgh, PA 15213. Phone: 412-623-7706; Fax: 412-623-7704. E-mail: lokshina@upmc.edu Copyright D 2005 American Association for Cancer Research. Cancer Epidemiology, Biomarkers & Prevention 981 Cancer Epidemiol Biomarkers Prev 2005;14(4). April 2005 Downloaded from http://aacrjournals.org/cebp/article-pdf/14/4/981/2264133/981-987.pdf by guest on 23 December 2022