Dataset Information

Age-Related Transcriptomic Profiling of Human Granulosa Cells Reveals mRNA-microRNA Regulatory Network Associated with Key Ovulation Dynamics [miRNA-Seq]

ABSTRACT: Advanced maternal age (AMA) patients experience decreased success from assisted reproductive technologies (ART), attributed to quantity and quality of oocytes, which is significantly influenced by the health of surrounding granulosa cells (GCs). In this study, we compared mRNA and microRNA (miRNA) transcriptomes between young (<32 y.o.) and AMA (>38 y.o.) patients' GCs to identify ovarian aging molecular signatures. Granulosa cells were isolated from 18 patients’ follicular fluid, and RNA was isolated for sequencing and subsequent bioinformatics analysis. We identified 293 and 21 differentially expressed genes (DEGs) and miRNAs (DE miRNA), respectively, between young and AMA GCs (P value < 0.05, FDR < 0.25, Fold Change > 1.5). Highly expressed mitochondrial-encoded genes, MT-ND3, MT-ND6, and MT-CYB are upregulated in young GCs. Pathway analysis indicates DEGs play a role in inflammation, cytokine signaling, extracellular matrix (ECM) remodeling, and angiogenesis. Some key DEGs include CXCL8, IL1B, NLRP3, SIGIRR, ANGPT2, ADAM8, and ADAMTS14. Additionally, target gene prediction and pathway analysis of DE miRNA indicates both up- and downregulated miRNA in young GCs are likely targeting genes associated with cell signaling, mitochondrial function, oxidative stress, apoptosis, and senescence pathways in addition to cytokine signaling, angiogenesis, and ECM remodeling. To further investigate regulatory mechanisms, we looked at the convergence of DEGs with predicted target genes of DE miRNA and identified miR-483-3p, miR-1268a, miR-4497, miR-7704, miR-135a-5p, miR-1261, and miR-4791 as key regulators of pathways involved with inflammation, ECM, and angiogenesis. The gene expression data suggests aged GCs have an impaired ability to elicit the same pro-inflammatory response combined with a dysregulation of angiogenesis and ECM remodeling when compared to young GCs, and miRNA may be playing a key role in the regulation of these key ovulatory events.

ORGANISM(S): Homo sapiens

PROVIDER: GSE270749 | GEO | 2025/03/12

REPOSITORIES: GEO

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Project description:To investigate the potential role of miRNAs in disease mechanisms, we conducted a thorough analysis by comparing the entire miRNome of 18 vEDS dermal fibroblasts to those of 18 control cells. To improve statistical reliability, we sequenced each sample from both groups as biological replicates. By applying a log2 fold change threshold of greater than 1 or less than -1, and an adjusted p-value of less than 0.05, we identified 137 DE-miRNAs in patient vs. control fibroblasts. Among these, 71 miRNAs exhibited decreased expression, while 66 miRNAs were upregulated. To explore the functional relationship between DE-miRNAs and the altered gene expression profile of vEDS cells, we conducted an extensive review of the literature and consulted several bioinformatics databases. Our search involved utilizing data from four miRNA-target prediction databases (microT-CDS, Targetscan, miRTarBase, and miRwalk) in addition to relevant literature sources providing information on the functional roles of these miRNAs. We thus focused our research on a specific subset of DE-miRNAs, known as miR-15b-5p, miR-16-5p, miR-21-3p, miR-24-3p, miR-29a-3p, miR-29b-3p, miR-138-5p, miR-145-5p, and miR-195-5p, which may play a biological role in dysregulated molecular mechanisms underlying disease pathogenesis. Our initial emphasis was on miR-29a-3p and miR-29b-3p due to their involvement in regulating critical cellular processes such as ECM remodeling into the vascular network, autophagy, cell proliferation, and apoptosis. To study how miR-29a-3p and miR-29b-3p contribute to the dysregulated cellular processes observed in patient cells, we performed GO and pathway enrichment analyses on their predicted target DEGs. GO analysis revealed that both these miRNAs may disturb fundamental biological functions, such as regulation of transcription by RNA polymerase II, negative gene expression regulation, miRNA-mediated gene silencing, translation regulation, heterochromatin organization, and ECM organization. This comphrensive transcriptome analysis of a large cohort of dermal fibroblasts from patients with vEDS reveals deregulated biological functions potentially involved in the disease pathogenesis. Taken together, the abnormal expression of several miRNAs suggests the presence of epigenetic control in vEDS cells, which could influence the transcriptional regulation of numerous target DEGs and related biological processes, potentially influencing disease mechanisms. This work was supported by Fondazione Telethon and the “Associazione con Giacomo contro vEDS”, (Grant number: GSA21F001, Seed Grant Fall 2021 vEDS) to Nicola Chiarelli.

Project description:Angiogenesis is a complex multicellular process requiring the orchestration of many events including migration, alignment, proliferation, lumen formation, remodeling, and maturation. Such complexity indicates that not only individual genes but also entire signaling pathways will be crucial in angiogenesis. To define an angiogenic blueprint of regulated genes, we utilized our well-characterized three-dimensional collagen gel model of in vitro angiogenesis, in which the majority of cells synchronously progress through defined morphological stages culminating in the formation of capillary tubes. We developed a comprehensive three-tiered approach using microarray analysis, which allowed us to identify genes known to be involved in angiogenesis and genes hitherto unlinked to angiogenesis as well as novel genes and has proven especially useful for genes where the magnitude of change is small. From a screen of miRNAs altered in in vitro angiogenesis we selected a subset that are predicted to target junctional molecules. MiR-27a, was rapidly downregulated upon stimulation of in vitro angiogenesis and its level of expression is reduced in neo-vessels in vivo. The downregulation of miR-27a was essential for angiogenesis since ectopic expression of miR-27a blocked capillary tube formation and angiogenesis. MiR-27a targets the junctional, endothelial specific cadherin, VE-cadherin. Consistent with this, vascular permeability to VEGF in mice is reduced by administration of a general miR-27 inhibitor. To determine that VE-cadherin was the dominant target of miR-27a function we used a novel technology with M-bM-^@M-^\BlockmirsM-bM-^@M-^], inhibitors that bind to the miR-27 binding site in VE-cadherin. The Blockmir CD5-2 demonstrated specificity for VE-cadherin and inhibited vascular leak in vitro and in vivo. Furthermore CD5-2 reduced oedema, increased capillary density and potently enhanced recovery form ischemic limb injury in mice. The Blockmir technology offers a refinement in the use of miRNAs especially for therapy. Further, targeting of endothelial junctional molecules by miRNAs has clinical potential especially in diseases associated with vascular leak. A microRNA microarray was performed to investigate miRNAs that were regulated during capillary tube formation and which could potentially be used as an M-bM-^@M-^XangiogenicM-bM-^@M-^Y miRNA profile. RNA was isolated from cells harvested at specific time points, previously identified as stages where major morphological changes are taking place18. These include 0.5 hour (invasion), 3 hours (migration), 6 hours (vacuole coalescence and lumen formation), 12 hours (lumen formation) and 24 hours (tube maturation).

Dataset Information

Age-Related Transcriptomic Profiling of Human Granulosa Cells Reveals mRNA-microRNA Regulatory Network Associated with Key Ovulation Dynamics [miRNA-Seq]

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets