Abstract
Coelomic fluid (CF) is the earliest dynamic and complex fluid of the gestational sac. CF contains maternal cells and proteins produced by embryonic cells, tissues and excretions. The biochemical composition of CF is modified throughout the first trimester of pregnancy and its protein profile reflects both physiological/pathological changes affecting the embryo and mother. Identification of variations in the balance of proteins might indicate particular types of pathologies, or ascertain specific genetic disorders. A platform utilizing protein enrichment procedures coupled with shotgun identification and iTRAQ differentiation provided the identification and quantitation of 88 unique embryonic proteins. It is relevant to note that chromosome X protein CXorf23 was found suggesting the embryo sex. Foetal sex was determined by Quantitative Fluorescent Polymerase Chain Reaction (QF-PCR) on coelomic cells, foetal tissues and maternal white blood cells, with a 100% concordance rate between iTRAQ-MS/MS and QF-PCR data. The functional associations among the identified proteins were investigated using STRING database. Open Targets Platform showed as significant the following therapeutic areas: nervous, respiratory, eye and head system disease.
Highlights
Coelomic fluid (CF) is the earliest fluid of the gestational sac, contained into the exocoelomic cavity (ECC) and it is in direct contact with placental villi during the first trimester of pregnancy[1]
To validate the results obtained by proteomic analysis, Quantitative Fluorescent Polymerase Chain Reaction (QF-PCR) amplification using several markers for chromosomes X, Y was performed on 22 serial samples of coelomic cells (CC), foetal tissue (FT) and maternal white blood cells (MBC)
CF is quite different in comparison to the other biological fluids which are usually adopted for clinical screening
Summary
Coelomic fluid (CF) is the earliest fluid of the gestational sac, contained into the exocoelomic cavity (ECC) and it is in direct contact with placental villi during the first trimester of pregnancy[1]. The application of various advanced MS-based platforms enabled the discovery of novel biomarkers in biological fluids[16] and tissues[17,18,19] Due to their high selectivity and high sensitivity, these MS-based platforms have been widely used for the identification and quantitation of metabolites[20,21], amino acids and their synthetic and non-natural analogues[22,23,24], medicinal materials, proteins in biological fluids and tissues. The presence of abundant serum proteins[33,36] represents a barrier to detection of medium and low abundance proteins in proteomic analyses This idea prompted us to develop three different analytical strategies for embryonic protein enrichment from normal CF (patients with no karyotype abnormalities) in order to increase the depth of CF proteome identification and to improve sensitivity for targeted analyses of differentially expressed proteins. OTP showed as significant the following therapeutic areas: “nervous system disease”, “respiratory system disease”, “eye disease” and “head disease”
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