Llano, RuthArdao Palacios, InésBrea Floriani, José ManuelRomero, LuzPata, María P.Martínez Rodríguez, Antón LeandroMacía Cortiñas, ManuelLoza García, María Isabel2025-07-072025-07-072025-06-17Llano, R.; Ardao, I.; Brea, J.M.; Romero, L.; Pata, M.P.; Martínez, A.L.; Macía, M.; Loza, M.I. Feasibility of Multiplex Cytokine Profiling in Preterm Labor: Towards Biomarker Discovery. Biology 2025, 14, 714.2079-7737https://hdl.handle.net/10347/42406Preterm delivery affects approximately 10% of pregnancies worldwide and remains a major clinical challenge due to the lack of reliable early predictive tools. Existing strategies are often invasive, relying on blood or amniotic fluid samples and requiring complex processing. In this study, we describe a novel non-invasive approach based on the multiplex detection of inflammatory cytokines in small urine volumes from pregnant women. To account for clinical and temporal variability, we applied Generalized Additive Models for Location, Scale, and Shape (GAMLSS) to adjust for gestational age at sampling and obstetric factors. Correlation network analyses revealed cytokine interactions that distinguished preterm from term deliveries, with macrophage-derived cytokines—MIP- 1α, MIP-1β, IL-15, and IL-22—emerging as central nodes. These findings highlight the involvement of the IL-1 pathway in the pathophysiology of preterm labor. Furthermore, urinary IL-5 and IL-31 levels correlated positively with pregnancy duration, whereas IL-1β and IL-1Ra in urine and TNFα in amniotic fluid showed inverse associations. Altogether, this non-invasive methodology provides insight into immune dynamics during pregnancy and offers a foundation for future studies focused on biomarker discovery and mechanistic understanding of preterm birth.eng© 2025 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license.Attribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/Preterm deliveryUrinary cytokinesCytokine networkInflammatory pathwaysIL-1 signalingGAMLSSLuminex multiplex technologyExploratory biomarker discovery241116 Fisiología de la reproducción2412 Inmunología1209 Estadísticajournal article10.3390/biology14060714open access