Single-cell transcriptomics of cultured skeletal stem cells with a PDGFRbeta D849V mutation
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE183404 | GEO | 2021/11/08
REPOSITORIES: GEO
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Project description:Spermatogonial stem cells are the foundation of spermatogenesis and as such can serve as a tool for the treatment of infertility in prepubertal cancer survivors. Spermatogonial stem cells are unique as they develop from primordial germ cells (PGCs), which colonize the developing tubules as immature SSC precursors. It has been controversial, when SSCs are maturing to an adult-like stem cell and recent research has found that prepubertal SSCs are actually metabolically distinct from adult SSCs until puberty. Sertoli cells picture a major part of the SSC niche and undergo drastic changes with puberty and polarize to compartmentalize the seminiferous epithelium with formation of tight junctions to a tight basal part where SSCs reside and an apical part with more differentiated stages of spermatogenesis. In the study were mapping the progression of Sertoli cells maturation events to the metabolic changes SSCs undergo during prepubertal development.
2023-02-24 | PXD037764 | Pride
Project description:Spermatogonial stem cells (SSCs) are the basis of spermatogenesis and therefore of male fertility. Transplantation of SSCs, isolated before treatment for cancer, and cultured in vitro, could be a potential treatment for infertility. Such clinical usage requires an understanding of the metabolic requirements during SSC development. Adult SSCs mainly use glycolysis for their maintenance in the mouse and human. However, SSCs embryonic precursors, primordial germ cells (PGCs), require a high mitochondrial metabolism in the mouse. Similarly, pig neonatal SSC precursors have been described to rely on oxidative phosphorylation (OXPHOS) for the first 2 months of development, when a transition to an adult SSC metabolic phenotype is initiated. When and if such a metabolic transition occurs in humans is ambiguous. We show here for the first time using single-cell RNA sequencing, that human PGCs and prepubertal human spermatogonia have an enrichment of oxidative phosphorylation associated genes, which is downregulated by 13 years of age. Furthermore, we show, that similar metabolic differences are detectable after birth also in mouse. The metabolic transition in humans with puberty was preceded by a drastic change of SSC shape. Using a pig model, we reveal that these metabolic changes could be regulated by IGF-1 dependent signaling via mTOR and proteasomic inhibition. Understanding the metabolic requirements of SSCs during development is crucial to establish a culture system and enable clinical use of SSCs.
2022-07-20 | GSE196801 | GEO
Project description:Spermatogonial stem cells (SSCs) are the basis of spermatogenesis and therefore of male fertility. Transplantation of SSCs, isolated before treatment for cancer, and cultured in vitro, could be a potential treatment for infertility. Such clinical usage requires an understanding of the metabolic requirements during SSC development. Adult SSCs mainly use glycolysis for their maintenance in the mouse and human. However, SSCs embryonic precursors, primordial germ cells (PGCs), require a high mitochondrial metabolism in the mouse. Similarly, pig neonatal SSC precursors have been described to rely on oxidative phosphorylation (OXPHOS) for the first 2 months of development, when a transition to an adult SSC metabolic phenotype is initiated. When and if such a metabolic transition occurs in humans is ambiguous. We show here for the first time using single-cell RNA sequencing, that human PGCs and prepubertal human spermatogonia have an enrichment of oxidative phosphorylation associated genes, which is downregulated by 13 years of age. Furthermore, we show, that similar metabolic differences are detectable after birth also in mouse. The metabolic transition in humans with puberty was preceded by a drastic change of SSC shape. Using a pig model, we reveal that these metabolic changes could be regulated by IGF-1 dependent signaling via mTOR and proteasomic inhibition. Understanding the metabolic requirements of SSCs during development is crucial to establish a culture system and enable clinical use of SSCs.
2022-07-20 | GSE196817 | GEO
Project description:Objective: Skeletal stem cells are known to be regulated by their local self-regulating niche, that is comprised mainly of their progeny and secreted paracrine factors. What cell intrinsic factors govern the maintenance and function of the SSC remains elusive. Methods: Using the previously characterized Tet1-/- mice we investigated the differences in skeletal stem cell lineage tree. Further we examined the ability of these cells to differentiate into osteogenic and chondrogenic lineages, both with and without injury. Transcriptomic differences and inflammatory cytokines were investigated for differences in cartilage development pathways. Results: Loss of Tet1 skews the SSC lineage tree by expanding the SSC pool at the expense of its progeny as well as by enhancing the chondrogenic differentiation of SSC and the multilineage bone cartilage stromal progenitors (BCSP). Tet1 inhibition leads to enhanced chondrogenesis in mouse BCSP upon injury, in human SSC and chondroprogenitors (CP) isolated from osteoarthritic (OA) human cartilage and in late stages of OA in a mouse model. Transcriptomic analyses of SSC and BCSP lacking Tet1 revealed dysregulated pathways including alterations in Tgfb signaling, melatonin degradation and cartilage development associated genes. Conclusion: Tet1 was identified as critical regulator of SSC fate. Loss of Tet1 selectively enhances chondrogenic ability of skeletal stem cells. Supplementation of melatonin can dampen inflammation and lead to better cartilage health.
2023-08-15 | GSE232666 | GEO
Project description:Because spermatogonial stem cells (SSCs) are virtually immortal by serial transplantation, SSC decrease in aged testes is considered to be caused by deteriorated microenvironment. Here we report cell-intrinsic mode of SSC aging. SSCs cultured for 5 years proliferate more actively than young SSCs and showed enhanced glycolytic activity but remain euploid and exhibited stable androgenetic imprinting patterns. Moreover, the aged SSCs kept constant SSC activity despite shortened telomeres. The increased proliferative activity of aged SSCs was caused by Wnt7b expression, which likely occurred due to decreased polycomb complex2 activity. Aberrant Wnt7b expression not only decreased reactive oxygen species levels and mitochondria activity but also stimulated glycolysis via c-jun N-terminal kinase (JNK) activation. Analysis of SSCs in Klotho aging mouse model also confirmed hyperactivation of JNK pathway and increased glycolysis. Therefore, not only SSC microenvironment but also intrinsic activation of Wnt7b-mediated glycolysis plays important roles in SSC aging.
2019-07-30 | GSE120777 | GEO
Project description:Because spermatogonial stem cells (SSCs) are virtually immortal by serial transplantation, SSC decrease in aged testes is considered to be caused by deteriorated microenvironment. Here we report cell-intrinsic mode of SSC aging. SSCs cultured for 5 years proliferate more actively than young SSCs and showed enhanced glycolytic activity but remain euploid and exhibited stable androgenetic imprinting patterns. Moreover, the aged SSCs kept constant SSC activity despite shortened telomeres. The increased proliferative activity of aged SSCs was caused by Wnt7b expression, which likely occurred due to decreased polycomb complex2 activity. Aberrant Wnt7b expression not only decreased reactive oxygen species levels and mitochondria activity but also stimulated glycolysis via c-jun N-terminal kinase (JNK) activation. Analysis of SSCs in Klotho aging mouse model also confirmed hyperactivation of JNK pathway and increased glycolysis. Therefore, not only SSC microenvironment but also intrinsic activation of Wnt7b-mediated glycolysis plays important roles in SSC aging.
2019-07-30 | GSE120775 | GEO
Project description:Antineoplastic treatments for cancer and other non-malignant disorders can result in male long-term or permanent infertility by ablating spermatogonial stem cells (SSCs). Preserving the future fertility of prepubertal patients faced with this prospect cannot rely on sperm banking. SSC transplantation using testicular tissue harvested before sterilizing treatment is a promising approach for restoring fertility, but lack of exclusive biomarkers which can unequivocally identify prepubertal SSCs in any primate species limits the therapeutic potential. To address this, we performed single-cell RNA-seq on unselected testis cells from immature baboons and macaques and compared these cells with published data from pre-pubertal human germ cells functionally-defined mouse SSCs. We found discrete groups of human spermatogonia while baboon and rhesus spermatogonia appeared less heterogenous. Cross-species analysis revealed cell types analogous to human SSCs in baboon and rhesus germ cells, but comparison with mouse SSCs revealed significant differences with primate SSCs. Indeed, a core component of a human SSC gene expression signature was conserved in baboon and rhesus spermatogonia and absent from mouse SSCs. These results further resolve the identity of pre-pubertal human SSCs and define novel pathways that could be leveraged for advancing their selection and propagation in vitro.
2023-05-15 | GSE222105 | GEO
Project description:We have used SmartSeq2 to sequence single phenotypic skeletal stem cell (SSCs) populations purified via FACS from adult mouse long bones. Two putative SSC populations were isolated based on their previously reported surface marker profiles. An osteochondrogenic SSC (ocSSC, CD45-CD31-Tie2-Ter119-CD51+6C3-Thy1-CD105-) and a perivascular SSC (pvSSC, CD45-CD31-Pdgfra+Sca1+CD24+) were investigated.
2021-06-11 | GSE161477 | GEO
Project description:We evaluated the degree to which gene expression patterns in primate skeletal cells differ across cell types and species. First, we optimized differentiation protocols of human and chimpanzee iPSCs through an intermediate MSC state and into chondrogenic or osteogenic cells. Single-cell RNA sequencing was collected from each stage of differentiation (iPSCs, MSCs, chondrogenic cells, and osteogenic cells) using the 10X Genomics platform to characterize gene expression variation.
2022-02-10 | GSE167240 | GEO
Project description:Maintenance and self-renewal of the spermatogonial stem cell (SSC) population is the cornerstone of male fertility. In this manuscript we have identified a key role for the nucleosome remodelling protein Chromodomain Helicase DNA binding protein 4 (CHD4) in regulating SSC function. Gene expression analyses revealed that CHD4 expression is largely restricted to spermatogonia in the mouse testis, and is particularly enriched in SSCs. Using spermatogonial transplantation techniques and RNAi mediated knockdown it was established that loss of Chd4 expression significantly impairs SSC regenerative capacity, resulting in a ~50% reduction in colonisation of recipient testes. A single cell RNA-seq comparison depicted reduced expression of ‘self-renewal’ genes such as Gfra1 and Pten following Chd4 knockdown, along with increased expression of signature progenitor genes, Neurog3 and Dazl. Co-immunoprecipitation analyses demonstrated that CHD4 regulates gene expression in spermatogonia not only though its traditional association with the remodelling complex NuRD, but also via interaction with the GDNF-responsive transcription factor SALL4. Cumulatively, the results of this study depict a previously unappreciated fundamental role for CHD4 in controlling fate decisions in the spermatogonial pool.
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