Transcriptome Landscape of Human Folliculogenesis Reveals Oocytes and Granulosa Cells Interactions
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PROVIDER: PRJNA421274 | ENA |
REPOSITORIES: ENA
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Project description:The dynamic transcriptional regulation and interactions of human germlines and surrounding somatic cells during folliculogenesis remains unknown. Using RNA-Seq analysis of human oocytes and corresponding granulosa cells (GCs) spanning five follicular stages, we revealed unique features in transcriptional machinery, transcription factor networks and reciprocal interactions in human oocytes and GCs that displayed developmental-stage-specific expression patterns. Notably, we identified specific gene signatures of two cell types in particular developmental stage that may reflect developmental competency and ovarian reserve. Additionally, we uncovered key pathways that may concert germline-somatic interactions and drive the transition of primordial-to-primary follicle which represents follicle activation. Thus, our work provides key insights into the crucial features of the transcriptional regulation in the stepwise folliculogenesis and offers important clues for improving follicle recruitment in-vivo and restoring fully competent oocytes in-vitro.
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Project description:Human ovarian folliculogenesis is an highly regulated and complex process. Characterization of follicular cell signatures during this dynamic process is important to understand follicle fate (to grow, become dominant or undergo atresia). The transcriptional signature of human oocytes and granulosa cells (GC) in early-growing and ovulatory follicles have been previously described, however that of oocytes with surrounding GCs in small antral follicles have not been studied yet. Here, we have generated a unique dataset of single-cell transcriptomics (SmartSeq2) consisting of the oocyte with surrounding GCs from several individual (non-dominant) small antral follicles isolated from adult human ovaries. We have identified two main types of (healthy) follicles, with a distinct oocyte and GC signature. Using the CellphoneDB algorithm, we then investigated the bi-directional ligand-receptor interactions regarding the TGFβ/BMP, WNT, NOTCH, and receptor tyrosine kinases (RTK) signaling pathways between oocyte and GCs within each antral follicle type. Our work not only revealed the diversity of small antral follicles but also contributes to fill the gap in mapping the molecular landscape of human folliculogenesis and oogenesis.
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Project description:Limited oocyte and ovarian reserve in vivo or chemo-therapy leads to reproductive aging or premature aging and associated diseases including infertility. Excitingly, oocytes have been successfully derived from embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) by ectopic expression of transcription factors, showing great potential in fertility preservation or restoration. The accessible granulosa cells are a type of somatic cells that interact and evolve with oocyte development during folliculogenesis. Further, with stem cells-like property, granulosa cells are amenable to reprogramming to generate iPSCs and have been the first used for clone animals. These prompted us to explore the potential of granulosa cells in derivation of germ cells. Meanwhile, the strict genome fidelity required for germ cells inspired us to test reprograming by complete small chemicals, which avoids genetic manipulation, cell transfection and destruction of embryos. Here we show that somatic granulosa cells of adult mouse ovaries can be converted to germ cells and functional oocytes that reproduce fertile pups. We are able to consistently induce granulosa cells to pluripotent state (gPSCs) like ESCs in both developmental competence and molecular signatures. Notably, crotonic sodium-facilitated crotonylation is critical not only for pure small chemicals-based reprogramming of granulosa cells to gPSCs, but also confers the gPSCs with high germline capacity. Consequently, the gPSCs and the derived primordial germ-cell like cells hold longer telomeres and maintain high genomic stability which is critical for germ cells. Taken together, we efficiently generate high quality gPSCs and functional oocytes from adult granulosa cells by significantly improving chemical reprograming approach.
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Project description:Separate transcription profiling of oocytes and granulosa cells for each follicle stage: primordial (PD), primary (PM), secondary (SC) follicles and the small antral stage (SA) obtained by Laser Capture Microdissection (LCM) and RNAseq. The purpose of this study was to describe global gene expression during early ovarian folliculogenesis for each follicular compartment, to identify differential and specific gene expression between the 2 follicular compartments and during follicular development, to investigate specific function and pathways and to explore bi-directional communication between oocytes and GC.
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Project description:Comparison of mitochondrial DNA (mtDNA) methylation patterns in oocytes, blastocysts and ovarian granulosa cells indicates hitherto unsuspected dynamics. Oocytes and blastocysts recovered from animals subjected to ovarian stimulation and from abattoir ovaries were analyzed using bisulphite transformation of DNA followed by whole genome sequencing. The number of mtDNA copies is quite high in oocytes (200–400,000) and early embryos, resulting in very high coverage (> 3000 X) and very low p values for each of 716 cytosine-based nucleosides. Methylation was lowest in oocytes, following by blastocysts then granulosa cells and was not restricted to CpG sites but found also at CHG and CHH sites not distributed randomly. Analysis of the corresponding genomic DNA also revealed numerous methylated CHG and CHH sites, particularly in oocytes. In association with ovarian stimulation, mtDNA methylation was lower in oocytes than in blastocysts, suggesting a developmental function. RNA analysis of mitochondria encoded genes showed a significant inverse correlation between methylation and expression level for all sequences. Methylation was more extensive in mature animals than in pre-pubertal animals and the profile in granulosa cells differed strikingly from those in oocytes and blastocysts. Although preliminary, the results suggest that mtDNA methylation might play a role during gametogenesis and might be subject to regulation according to animal environment and/or age.
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