Transcriptomic landscape of cumulus cells from patients <38 years old with a history of poor ovarian response (POR) treated with platelet-rich plasma (PRP)

Abstract

Intraovarian injection of autologous platelet-rich plasma (PRP) has recently been investigated as a potential treatment for patients with diminished ovarian reserve. In the current study, differential gene expression in cumulus cells obtained from patients treated with PRP was compared to controls. RNA sequencing libraries were constructed from the cumulus cells, and differential expression analysis was performed with a false discovery rate threshold of p-value ≤0.05 and Log2 fold change ≥0.584. RNA sequencing of cumulus cells revealed significant differences in gene expression when comparing those treated with PRP and resulted in a live birth (n = 5) to controls with live birth (n = 5), or to controls with failed implantation (n = 5). Similarly, when all samples treated with PRP (those that resulted in live birth or arrested embryos (n = 10)) were compared to all samples from controls (those that resulted in live birth, no pregnancy, or arrested embryos (n = 13)), gene expression was significantly different. Several pathways were consistently affected by PRP treatment through multiple comparisons, including carbohydrate metabolism, cell death and survival, cell growth and proliferation, and cell-to-cell signaling, all of which have been implicated in human causes of infertility.

Keywords: RNA-sequencing; diminished ovarian reserve; plasma rich protein.

Figure 1

Gene expression is altered in PRP-treated patients with sustained implantation (Group 2) compared to control patients with sustained implantation (Group 1). (A) The heat map illustration shows differentially expressed genes. The color spectrum ranging from red to blue indicates normalized levels of gene expression from high to low. (B) Volcano plot for RNA-seq comparing Group 2 to Group 1. (C) Differentially expressed genes in Group 2 compared to Group 1, P < 0.05 for each. For the box plots, the bottom and top whiskers denote 5 and 95 percentile values, the bottom and top bounds of the rectangle denote the 25 and 75 percentile values, and the line in between denotes the median (50 percentile) value of the distribution. (D) PCA plot for RNA-seq for significant genes comparing Group 2 to Group 1. The transcripts per million (TPM) value represents the relative expression level comparable between samples.

Figure 2

Pathway analysis comparing patients with sustained implantation (Group 2) to control patients with sustained implantation (Group 1). Pathway analysis was performed using the Gene Ontology bioinformatics tool. Log2 fold change (FC) ≥0.584 false discovery rate (FDR).

Figure 3

Gene expression is altered in cumulus cells of PRP-treated patients (Groups 2 and 5 combined) compared to controls (Groups 1, 3, and 4 combined). (A) The heat map illustration shows differentially expressed genes. The color spectrum ranging from red to blue indicates normalized levels of gene expression from high to low. (B) Volcano plots for RNA-seq comparing PRP to control. (C) Differentially expressed genes in CONT and PRP, P < 0.05 for each. For the box plots, the bottom and top whiskers denote 5 and 95 percentile values, the bottom and top bounds of the rectangle denote the 25 and 75 percentile values, and the line in between denotes the median (50 percentile) value of the distribution. (D) PCA plots for RNA-seq for significant genes comparing PRP to control. The transcripts per million (TPM) value represents the relative expression level comparable between samples.

Figure 4

Pathway analysis comparing patients treated with PRP (Groups 2 and 5 combined) to controls (Groups 1, 3, and 4 combined). Pathway analysis was performed using the Gene Ontology bioinformatics tool. Log2 fold change (FC) ≥0.584 false discovery rate (FDR).

Figure 5

Gene expression is altered in PRP-treated patients with sustained implantation (Group 2) compared to control patients without sustained implantation (Group 3). (A) The heat map illustration shows differentially expressed genes. The color spectrum ranging from red to blue indicates normalized levels of gene expression from high to low. (B) Volcano plots for RNA-seq comparing PRP-LB with C-NP. (C) Differentially expressed genes in Group 2 versus Group 3, P < 0.05 for each. For the box plots, the bottom and top whiskers denote 5 and 95 percentile values, the bottom and top bounds of the rectangle denote the 25 and 75 percentile values, and the line in between denotes the median (50 percentile) value of the distribution. (D) PCA plots for RNA-seq for significant genes comparing Group 2 with Group 3. The transcripts per million (TPM) value represents the relative expression level comparable between samples.

Figure 6

Pathway analysis comparing patients with sustained implantation (Group 2) to control patients without sustained implantation (Group 3). Pathway analysis was performed using the Gene Ontology bioinformatics tool. Log2 fold change (FC) ≥0.584 false discovery rate (FDR).