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EID1 knochdown by small RNA induces the large-scale conversion of adipose-derived mesenchymal stem cells into hematopoietic stem cells

ABSTRACT: Comparison of expression profiles in adipose derived MSCs (AD-MSCs) without or with transfection of siR-EID1 and shR-EID1 and cord blood-derived HSCs (CB-HSCs). After MSCs were transfected with sRNA-EID1, they could be converted into HSCs. The goal was to determine possible molecular mechanisms of MSC transdetermination.

ORGANISM(S): Homo sapiens

PROVIDER: GSE42650 | GEO | 2012/12/01

SECONDARY ACCESSION(S): PRJNA182511

REPOSITORIES: GEO

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EID1 knochdown by small RNA induces the large-scale conversion of adipose-derived mesenchymal stem cells into hematopoietic stem cells

Project description:Comparison of expression profiles in adipose derived MSCs (AD-MSCs) without or with transfection of siR-EID1 and shR-EID1 and cord blood-derived HSCs (CB-HSCs). After MSCs were transfected with sRNA-EID1, they could be converted into HSCs. The goal was to determine possible molecular mechanisms of MSC transdetermination. Two-condition experiments: AD-MSCs vs CB-HSCs, and AD-MSCs transfected with the combination of siR-EID1 and shR-EID1 vs AD-MSCs transfected with shR-EID1.

2012-12-01 | E-GEOD-42650 | biostudies-arrayexpress

Directional capacity of human mesenchymal stem cells to support hematopoietic stem cell proliferation in vitro

Project description:Objectives: Hematopoietic stem cells (HSCs) reside in a specialised microenvironment in the bone marrow, which is majorly composed of mesenchymal stem cells (MSCs) and its' derivatives. This study aimed to investigate the regulatory role of MSCs to decipher the cellular and humoral communications on HSCs' proliferation, self-renewal, and differentiation at the transcriptomic level. Materials and methods: Microarray assay was employed to analyse the gene expression profile of HSCs that imparted by MSCs during co-culture. Results: The proliferation of human umbilical cord blood-derived HSCs (hUC-HSCs) markedly propagated when MSCs were used as the feeder layer, without disturbing the undifferentiated state of HSCs, and reduced the cell death of HSCs. Upon co-culture with MSCs, the global microarray analysis of HSCs disclosed 712 differentially expressed genes (DEGs) (561 up-regulated and 151 down-regulated). The dysregulations of various transcripts were enriched for cellular functions such as cell cycle (including CCND1), apoptosis (including TNF), and genes related to signalling pathways governing self-renewal, as well as WNT5A from the Wnt signalling pathway, MAPK, Hedgehog, FGF2 from FGF, Jak-STAT, and PITX2 from the TGF-β signalling pathway. To concur this, real-time quantitative PCR (RT-qPCR) was utilised for corroborating the microarray results from five of the most dysregulated genes. Conclusion: This study elucidates the underlying mechanisms of the mitogenic influences of MSCs on the propagation of HSCs. The exploitation of such mechanisms provides a potential means for achieving larger quantities of HSCs in vitro, thus obviating the need for manipulating their differentiation potential for clinical application.

2022-05-28 | GSE176002 | GEO

Bmi1 suppresses adipogenesis in the hematopoietic stem cell niche: ChIP-Seq

Project description:Bone marrow mesenchymal stromal cells (MSCs) that express high levels of stem cell factor (SCF) and CXC chemokine ligand 12 (CXCL12) are one crucial component of the hematopoietic stem cell (HSC) niche. While the secreted factors produced by MSCs to support HSCs have been well described, little is known regarding the transcriptional regulators controlling the cell fate of MSCs and thus indirectly maintaining HSCs. Bmi1 is a polycomb group protein that regulates HSCs both cell intrinsically and extrinsically, but it is unknown in which cell type and how Bmi1 functions to maintain HSCs extrinsically. Here we show that Bmi1 maintains HSCs by preventing adipogenic differentiation of MSCs. Bmi1 is highly expressed in MSCs but becomes downregulated upon adipogenic differentiation and during aging. Deleting Bmi1 from MSCs increased marrow adipocytes, induced HSC quiescence and depletion, and impaired hematopoiesis. We found that Bmi1 repressed multiple developmental programs in MSCs by safeguarding the repressive epigenetic marks histone H2A ubiquitylation and H3 lysine 27 trimethylation. We identified a novel adipogenic program governed by Pax3, which Bmi1 repressed in MSCs. Our results establish Bmi1 as a critical regulator of MSC cell fate that suppresses marrow adipogenesis to create a supportive niche for HSCs.

2019-04-12 | GSE121289 | GEO

Bmi1 suppresses adipogenesis in the hematopoietic stem cell niche: RNA-Seq

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RNA sequencing analyisis to identify genes regulated by GW9662 in human cord blood hematopoietic stem and progenitor cells

Project description:PPARγ antagonist GW9662 treatment could enhance ex vivo expansion of human cord blood hematopoietic stem and progenitor cells (HSCs/HPCs). To gain mechanistical insights into how antagonizing PPARγ promotes expansion of HSCs/HPCs, we performed RNA sequencing (RNA seq) analysis to identify genes involved in this process. Loss of function of PPARγ in CB CD34+ cells resulted in downregulation of a number of differentiation associated genes, including CD38, CD1d, HIC1, FAM20C, DUSP4, DHRS3 and ALDH1A2, suggesting that PPARγ antagonist may maintain stemness of CB CD34+ cells, at least in part by preventing differentiation. We also observed that FBP1, encoding fructose 1, 6-bisphosphatase, a negative regulator of glycolytic flux, was significantly downregulated by treating CB CD34+ cells with GW9662. Our study demonstrates that antagonizing PPARγ signaling drives ex vivo expansion of human CB HSCs/HPCs by switching on FBP1 repressed glycolysis and preventing differentiation.

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miR-125a Confers Multi-Lineage Long-Term Repopulating Stem Cell Activity to Murine and Human Hematopoietic Progenitors

Project description:Umbilical cord blood (CB) is a non-invasive, convenient and broadly used source of hematopoietic stem cells (HSCs) for allogeneic stem cell transplantation. However, limiting numbers of HSCs remain a major constraint for its clinical application. One feasible option would be to expand HSCs to improve therapeutic outcome, however available protocols and the molecular mechanisms governing the self-renewal of HSC are unclear. Here we show that ectopic expression of a single miRNA, miR-125a, in purified murine and human multipotent progenitors (MPP) resulted in increased self-renewal and robust long-term multi-lineage repopulation in transplanted recipient mice. Using quantitative proteomics and Western blot analysis, we identified a restricted set of miR-125a targets which revealed the involvement of the MAP kinase signaling pathway in conferring long-term repopulating capacity to multipotent progenitors in human and mice. Our findings offer the innovative potential to use MPP with enhanced self-renewal activity to augment limited sources of HSC to improve clinical protocols.

2017-01-11 | PXD003931 | Pride

MicroRNA expression profile of human hepatic stellate cells LX-2 co-cultured with human umbilical cord mesenchymal stem cells and control group

Project description:Hepatic stellate cells(HSCs) are the main effector cells of liver fibrosis. In order to study the effect of mesenchymal stem cells(MSCs) on microRNAs expression of HSCs, we co-cultured HSCs LX-2 activated by TGFβ1 with human umbilical cord MSCs(hUC-MSCs) for 48 hours, and compared the differentially expressed miRNA with LX-2 cultured alone by high-throughput sequencing. The results showed that two mature microRNAs expressed increased, and nine expressed decreased.

2020-05-24 | GSE151098 | GEO

Transcriptional and protein-level profiling of engraftable human fetal liver (fl)-derived hematopoietic stem cells (HSCS)

Project description:We provide an in-depth characterization of engraftable fetal liver (FL) HSCs as these have been shown to possess superior engraftment potential compared to cord blood (CB) and bone marrow (BM) HSCs. We have single cell profiled 26,407 FL cells, of which 7,235 highly-enriched functional FL HSCs, both at the transcriptional and protein level to uncover the detailed molecular signature of engraftment potential.

2021-02-08 | GSE160251 | GEO

Time-course analysis of transcriptome changes induced by ex vivo culture in human cord blood hematopoietic stem cell at single cell resolution [CB]

Project description:Here we present the single cell transcriptomes of human cord blood (CB) long-term hematopoietic stem cells (LT-HSCs) cultured in differentiation promoting medium over a time-course of 0, 6 24 or 72 h. A condition was included for LT-HSCs cultured with the CDK6 inhibitor Palbociclib for durations of 24 or 72 h. Palbociclib causes reversible cell cycle arrest in early G1 in these culture conditions. The objectives of this study were to understand the impact of cell culture on CB LT-HSCs in a time resolved manner and to investigate the role of cell cycle progression in driving this transcriptional change. In the present study we report that 72 h of culture causes extensive rewiring of the CB LT-HSC transcriptome and that changes are dynamically acquired over the time-course. We also report that cell cycle progression is not the main driver of transcriptional change in tested culture conditions.

2022-11-19 | GSE213365 | GEO

Hematopoietic-mesenchymal signal regulates properties of mesenchymal stem cells

Project description:It is well known that the properties of hematopoietic stem cells (HSCs) such as their ability to self-renew, and multipotency are maintained through interactions with mesenchymal cells. Meanwhile, the identification of cells that are the key component of niche remains a subject of dispute. Mesenchymal stem cells (MSCs) are one of the niche component cells, MSCs are one of the niche component cells which provide stemness signal to HSCs, while it is also hypothesized that there is also a counterpart signal from hematopoiesistic cells to mesenchymeal cells, as HSCs also constitute core stem cells niche for MSCs. Although mesenchymal stem cells (MSCs) have also been studied intensively, the signals involved in the maintenance of their properties have not attracted much attentions. We used microarrays to detail the global programme of gene expression underlying HSC-MSC interaction and identified distinct classes of regulated genes during this process.

2021-12-31 | GSE186250 | GEO

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