Quantitative proteomic analysis of sperm in unexplained recurrent pregnancy loss

Abstract

Background Recurrent pregnancy loss (RPL) refers to two or more spontaneous abortions that occur consecutively with the same spouse. RPL severely affects human reproduction health, and causes extreme physical and mental suffering to patients and their families. Methods We used isobaric tags for relative and absolute quantitation (iTRAQ), which was coupled with liquid chromatography mass spectrometry (LC-MS) proteomic analysis, in order to identify differentially expressed proteins. Moreover, we used western blot to analyze differentially expressed proteins. Results Of the 2350 non-redundant proteins identified, 38 proteins were significantly altered and were identified as potential biomarkers for RPL. The protein-protein interaction network constructed using GeneMANIA revealed that 35.55% displayed similar co-expression, 30.87% were predicted, and 20.95% had physical interaction characteristics. Based on Gene ontology classification and KEGG pathway enrichment analyses, the majority of these differentially expressed proteins were found to be related to biological regulation, metabolic and cellular processes, protein binding and different enzymes activities, as well as disorder of fat and glucose metabolic pathways. It is noteworthy that three metabolism related biomarkers (HK1, ACLY, and FASN) were further confirmed through western blot analysis. Conclusions These results suggest that these differentially expressed proteins may be used as biomarkers for RPL, and related signaling pathways may play crucial roles in male induced RPL. Electronic supplementary material The online version of this article (10.1186/s12958-019-0496-5) contains supplementary material, which is available to authorized users.

TMT-Based Proteomic Analysis of Human Spermatozoa from Unexplained Recurrent Miscarriage Patients before and after Oral Antioxidant Treatment

“…Comparative proteomics studies have previously been conducted on sperm [ 29 , 30 , 31 ]. However, RM patients have only been included in a few recent reports [ 23 , 32 ]. To our knowledge, this is the first study that analyzes the protein profile of patients categorized as being at high risk of RM according to their sperm DNA DSB profile.…”

“…Despite the advances in the search for male-factor-related biomarkers for RM, such as the presence of DSB, new molecular biomarkers need to be identified in order to better understand the physiopathology of this disease. In this sense, a study by Xue et al [ 23 ] identified 38 proteins that were significantly altered in RM patients, mostly related to metabolic processes, biological regulation, and protein binding activities [ 23 ]. Additionally, a study by Sutovsky et al [ 24 ] identified that patients with high SPTRX3 protein content in semen are prone to RM [ 24 ].…”

Proteomic Profile of Sperm in Infertile Males Reveals Changes in Metabolic Pathways

“…In addition, the iTRAQ-based proteomic technology that offers improved accuracy in terms of protein quantification was used. The methods used to screen differentially expressed proteins in the two previous studies included LC-MS/MS, two-dimensional gel electrophoresis, liquid chromatography-mass spectrometry, and matrixassisted laser desorption ionization-time-of-flight-mass spectrometry analysis; however, the iTRAQ-based technology used here is a high-throughput method used extensively for quantitative proteomic analysis of sperm [7,20,21]. Therefore, as expected, additional differentially expressed proteins that are potentially associated with infertility were discovered.…”

In Silico Sperm Proteome Analysis to Investigate DNA Repair Mechanisms in Varicocele Patients

“…This protein has seven catalytic activities and a site for the binding of the prosthetic group 4'-phosphopantetheine of the acyl carrier protein (ACP) domain (51). Overexpression of FASN was confirmed by WB analysis in semen samples from couples with unexplained recurrent pregnancy loss (52). In somatic cells, homologous recombination (HR) is the main molecular mechanism for DSB repair, which occurs by using the sister chromatid as a template.…”