title: An integrated approach based on multiplexed protein array and iTRAQ labeling for in-depth identification of pathways associated to IVF outcome.
abstract: The emergence of high-throughput protein quantification methodologies has enabled the comprehensive characterization by longitudinal and cross-sectional studies of biological fluids under physiological and pathological conditions. In particular, the simultaneous investigation of cytokines and growth factors signaling pathways and their associated downstream effectors by integrated multiplexed approaches offers a powerful strategy to gain insights into biological networks and processes in living systems. A growing body of research indicates that bioactive molecules of human reproductive fluids, including human follicular fluid (hFF), may affect oocyte quality, fertilization and embryo development, thus potentially influencing the physiopathology of pregnancy-related conditions. In this work, an iTRAQ labeling strategy has been complemented with a multiplexed protein array approach to analyze hFFs with the aim to investigate biological processes and pathways related to in vitro fertilization (IVF) outcome. The iTRAQ labeling strategy lead to the quantification of 89 proteins, 30 of which were differentially expressed in hFFs with successful compared to unsuccessful IVF outcome. The targeted study, based on multiplexed antibody protein arrays, allowed the simultaneous quantification of 27 low abundance proteins, including growth factors, chemokines and cytokines endowed with pro- and anti-inflammatory activity. A significant number of differentially regulated proteins were involved in biological functions related to blood coagulation, acute phase response signaling and complement system. Overall, the present results provide an integrated overview of protein changes in hFFs associated to IVF outcome, thus improving current knowledge in reproductive medicine and fertility research.
authors.affiliation: Department of Environmental, Biological and Pharmaceutical Sciences and Technologies, Second University of Naples, Caserta, Italia ; Institute of Biostructures and Bioimaging-IBB, CNR, Napoli, Italia ; CIRPEB-Centro Interuniversitario di Ricerca sui Peptidi Bioattivi, Napoli, Italia.
authors.forename: Valeria
authors.initials: V
authors.lastname: Severino
authors.forename: Livia
authors.initials: L
authors.lastname: Malorni
authors.forename: Anna Emilia
authors.initials: AE
authors.lastname: Cicatiello
authors.forename: Vittoria
authors.initials: V
authors.lastname: D'Esposito
authors.forename: Salvatore
authors.initials: S
authors.lastname: Longobardi
authors.forename: Nicola
authors.initials: N
authors.lastname: Colacurci
authors.forename: Nadia
authors.initials: N
authors.lastname: Miraglia
authors.forename: Nicola
authors.initials: N
authors.lastname: Sannolo
authors.forename: Annarita
authors.initials: A
authors.lastname: Farina
authors.forename: Angela
authors.initials: A
authors.lastname: Chambery
mesh_terms: D019295:Computational Biology; D003430:Cross-Sectional Studies; D016207:Cytokines / Q000378:metabolism; D005260:Female; D005307:Fertilization in Vitro; D015571:Follicular Fluid / Q000378:metabolism*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D009865:Oocytes / Q000378:metabolism*; D011247:Pregnancy; D011256:Pregnancy Outcome; D040081:Protein Array Analysis / Q000379:methods; D020543:Proteome; D040901:Proteomics / Q000379:methods; D015203:Reproducibility of Results; D015398:Signal Transduction
chemical_list: D016207:Cytokines; D036341:Intercellular Signaling Peptides and Proteins; D020543:Proteome