Abstract


OBJECTIVE: To evaluate if transcriptomic variations in the endometrium between women with of RIF and fertile control during window of implantation reveal insight to the etiology of RIF.
DESIGN: In this prospective cohort study, mRNA fractions were extracted from 45 endometrial biopsies obtained from women with RIF (n=24) or fertile controls (n=21) during the window of implantation (LH+7 to LH+10). Endometrial biopsies were collected using a Pipelle. Total RNA and protein were isolated from endometrium tissue samples. view more

OBJECTIVE: To evaluate if transcriptomic variations in the endometrium between women with of RIF and fertile control during window of implantation reveal insight to the etiology of RIF.
DESIGN: In this prospective cohort study, mRNA fractions were extracted from 45 endometrial biopsies obtained from women with RIF (n=24) or fertile controls (n=21) during the window of implantation (LH+7 to LH+10). Endometrial biopsies were collected using a Pipelle. Total RNA and protein were isolated from endometrium tissue samples.
MATERIALS AND METHODS: mRNA extracted from 45 endometrial biopsies obtained from women with RIF (age ≤38 years and BMI ≤28) (n=24) or fertile controls (age and BMI matched) (n=21) during the window of implantation (LH+7 to LH+10) and analyzed using Agilent/SurePrint G3 Human GE2 8x60k microarrays. The data was preprocessed and analyzed with the R package limma. RIF patients had a history of implantation failure from at least three consecutive IVF attempts in which 2-3 embryos of high grade quality were transferred in every cycle. Fertile controls had a history of at least one live birth.
RESULTS: In this study, we have applied a genome-wide transcript-level changes approach using oligo microarrays representing 37,600 genes to define the transcriptomic profile of RIF patients in comparison to fertile controls.The differentially-expressed genes (DEGs) that met the criteria | log2(Fold Change) | (LFC)≥1 and adjusted p-value≤0.05 were accepted as statistically significant DEGs. The resulting 641 DEGs that met the criteria were then used for further selection with a feature selection wrapper algorithm that helps in identifying aspects the most relevant to the distinction between patients and controls. Feature selection was performed 100 times and DEGs that were chosen in all runs were accepted to be relevant DEGs. Functional enrichment analysis was carried out with 93 DEGs that were accepted as relevant. The BIOCARTA pathway “RAC1 cell motility signaling pathway” as well as related pathways were found to be enriched.
CONCLUSIONS: It is becoming evident that endometrial cells are inherently invasive and probably contribute to the processes at the implantation site. In this study, RAC1 was shown to be involved in endometrial stromal cell migration, which affects embryo implantation. Local growth factors such as PDGF-BB and HB-EGF may be utilized to enhance migration at the implantation site of patients with RIF.

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