Female unexplained infertility is a multifactorial disorder with poorly understood pathogenesis with underlying mechanisms. Around 50% of infertility cases are caused solely by female factors, with 15-20% having no identifiable cause, making it a primary challenge for reproductive medicine experts for decades. Among all omic procedures, proteomics is now being widely used in the field of human reproduction. In recent years, the literature on the proteome database connected to various reproductive tissues in males and females has been enhanced by the availability of improved proteomic methods and databases. Further, Network and pathway analysis using bioinformatic tools has now provided a more scholastic and comprehensive view of the potential pathways connected to the significantly elevated DEPs (Differentially Expressed Proteins) to identify their relevance to specific infertility circumstances. This study investigated differential tissue proteome profiles in patients with unexplained female infertility and healthy fertile controls and explored the association between their tissue proteomes. Paired samples of ten patients with unexplained infertility and ten healthy fertile controls during their mid-secretory phase (LH +7) were obtained for optimized quantitative tissue proteomics analysis. High-resolution two-dimensional gel electrophoresis (2-DE) analysis was used for comparing patterns of protein expression, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry was used to identify protein spots on 2-DE gels (MALDI-TOF MS). The presence of upregulated DEPs were then validated against their respective primers using the qPCR technique. To interpret and analyze the collected data in light of their biological function, pathway, and interaction network, bioinformatic software and tools, including UniProt, Genecards (v.4.8.2), KEGG, Reactome, and STRING were utilized (version 10.5). Using PANTHERS 14.0, the genes corresponding to differentially expressed proteins were mapped to multiple gene annotation data and the biological pathway. Molecular function, biological process, cellular location, and protein class was done. We identified twenty four proteins differentially expressed in Infertility when compared with healthy controls, including Ankyrin repeat domain-containing protein 36A (ANKRD36), Zinc finger protein 658 (ZNF658B), MAM and LDL receptor class A domain containing 1(MALRD 1) and proline-rich coiled-coil 2A (PRRC2A). There appears to be an interesting strong association among the genes of three metabolic enzymes: between Zinc finger CCCH domain-containing protein 13 and Putative RNA-binding protein 15 and between homologues of Hemoglobin subunit beta- HBB and HBA1 (combined score: 0.781), HBB and HBA2 (combined score: 0.560) and HBA1 and HBA2 (combined score: 0.800). In addition, PRRC2B co-expresses with ERCC6, CNTLN co-expresses with ANKRD36; ZC3H1D co-expresses with EIF2AK2; CAMSAP2 co-expresses with EIF2AK2; and EIF2AK2 coexpresses with LAMA3 not only an interaction between EIF2AK2, CAMSAP2, LAMA3 and ANKRD36 but also a possible association between them and unexplained female infertility was noted. The enrichment study identified 25, 20, and 114 pathways among KEGG, WiKi, and Reactome databases, respectively indicating the key role these proteins play in causing the morbidity associated with infertility. Further understanding of the underlying causes and the mechanistic pathways of unexplained female infertility is needed to better comprehend the disease and its management.
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