Abstract 5298: Multiplex immunoassay characterization of 48 cytokines, chemokines, and growth factors in colorectal cancer

Abstract

Abstract Cytokines, chemokines, and growth factors are critical mediators of immune system function capable of signaling through autocrine, paracrine, and endocrine mechanisms. Their pleiotropic immunomodulatory properties allow these biomolecules to react to diverse stimuli and regulate the immune response. We developed a novel immunoassay, using Luminex xMAP technology, for the simultaneous measurement of 48 cytokines, chemokines, and growth factors in serum, plasma, and cell culture supernatant. Many of these proteins have been proposed as biomarkers of colorectal cancer (CRC) and as key regulators of the tumor microenvironment (TME). While this complex milieu can support antitumor immunity, CRC is frequently characterized by chronic inflammation driving disease progression, with the TME serving as a potent reservoir from which the cytokines, chemokines, and growth factors exert their influence. The MILLIPLEX Human Cytokine/Chemokine/Growth Factor Panel B (HCYTB-60K) assays were performed in 96-well plates and run on the Luminex LX200™ instrument. Data was acquired via xPONENT v. 4.3 software. Data analysis was performed for all immunoassays using the Belysa Immunoassay Curve Fitting Software. Commercially available serum samples obtained from colorectal cancer patients and healthy controls were evaluated for cytokine, chemokine, and growth factor expression. Of the 48 analytes tested, statistically significant increases in BAFF, sFas, SCF, and MPIF-1 were observed in colorectal cancer serum. In contrast, IL-20 and ENA-78 decreased relative to healthy controls. While the immunoassay was verified for measurement of serum and plasma samples, additional sample types may be of interest to cancer researchers. Therefore, we evaluated the ability of the 48-plex immunoassay to measure these biomarkers in CSF, urine, milk, tumor samples and exosomal isolates, establishing a profile for these proteins in each sample type. Altogether, this study uses a novel multiplex immunoassay to characterize the immunological profile of colorectal cancer serum, focusing on 48 cytokines, chemokines, and growth factors. Additionally, alternative sample types were evaluated to establish the utility of this technology for applications beyond serum, plasma, and cell culture supernatant. Citation Format: Wen-Rong Lie, Harold Steiner, Sasha Williams, Munmun Banerjee, Anthony Saporita, Brooke Gilliam, Qiang Xiao. Multiplex immunoassay characterization of 48 cytokines, chemokines, and growth factors in colorectal cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5298.