title: The search for biomarkers of human embryo developmental potential in IVF: a comprehensive proteomic approach.
abstract: The objective of these studies was to identify differentially expressed peptides/proteins in the culture media of embryos grown during in vitro fertilization (IVF) treatment to establish their value as biomarkers predictive of implantation potential and live birth. Micro-droplets of embryo culture media from IVF patients (conditioned) and control media maintained under identical culture conditions were collected and frozen at -80°C on Days 2-3 of in vitro development prior to analysis. The embryos were transferred on Day 3. The peptides were affinity purified based on their physico-chemical properties and profiled by mass spectrometry for differential expression. The identified proteins were further characterized by western blot and ELISA, and absolute quantification was achieved by multiple reaction monitoring (MRM). We identified up to 14 differentially regulated peptides after capture using paramagnetic beads with different affinities. These differentially expressed peptides were used to generate genetic algorithms (GAs) with a recognition capability of 71-84% for embryo transfer cycles resulting in pregnancy and 75-89% for those with failed implantation. Several peptides were further identified as fragments of Apolipoprotein A-1, which showed consistent and significantly reduced expression in the embryo media samples from embryo transfer cycles resulting in viable pregnancies. Western blot and ELISA, as well as quantitative MRM results, were confirmatory. These results demonstrated that peptide/protein profiles from the culture medium during early human in vitro development can discriminate embryos with highest and lowest implantation competence following uterine transfer. Further prospective studies are needed to establish validated thresholds for clinical application.
authors.affiliation: Department of Microbiology and Molecular Cell Biology, Center for Biomedical Proteomics, Eastern Virginia Medical School, Norfolk, VA 23507, USA.
authors.forename: Julius
authors.initials: J
authors.lastname: Nyalwidhe
authors.forename: Tanya
authors.initials: T
authors.lastname: Burch
authors.forename: Silvina
authors.initials: S
authors.lastname: Bocca
authors.forename: Lisa
authors.initials: L
authors.lastname: Cazares
authors.forename: Shamina
authors.initials: S
authors.lastname: Green-Mitchell
authors.forename: Marissa
authors.initials: M
authors.lastname: Cooke
authors.forename: Paige
authors.initials: P
authors.lastname: Birdsall
authors.forename: Gaurav
authors.initials: G
authors.lastname: Basu
authors.forename: O John
authors.initials: OJ
authors.lastname: Semmes
authors.forename: Sergio
authors.initials: S
authors.lastname: Oehninger
mesh_terms: D000328:Adult; D016632:Apolipoprotein A-I / Q000737:chemistry / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D017077:Culture Media, Conditioned; D004622:Embryo, Mammalian / Q000378:metabolism*; D047108:Embryonic Development; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005307:Fertilization in Vitro; D006801:Humans; D008875:Middle Aged; D011247:Pregnancy; D011256:Pregnancy Outcome; D040901:Proteomics; D012189:Retrospective Studies; D053719:Tandem Mass Spectrometry
chemical_list: D016632:Apolipoprotein A-I; D015415:Biomarkers; D017077:Culture Media, Conditioned