Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Ex-vivo Human Hematopoietic Stem Cell Expansion Requires Coordination of Cellular Reprogramming with Mitochondrial Remodeling and P53 Activation [bulk]

ABSTRACT: Ex-vivo culture conditions used to expand the numbers of hematopoietic stem cells (HSCs) present within an umbilical cord blood (UCB) unit create cellular stresses leading to loss or at best maintenance of the primitive HSCs. Recently, we have shown that treatment with a histone deacetylase inhibitor, valproic acid (VPA), increases substantially the numbers of transplantable HSCs from UCB-CD34+ cells. In this report, we demonstrate that VPA treatment orchestrates cellular mechanisms that drive UCB-CD34+ cells into a primitive state in which they acquire and retain phenotypic, transcriptomic and primitive mitochondrial profiles, all of which characterize long-term HSCs. Remarkably, our data link the acquisition of the HSC phenotype to the remodeling of the mitochondrial network and p53 activation and establish both as critical regulators of ROS and therefore of HSC fate. VPA treatment leads to restructured mitochondria with reduced mass, membrane potential and ROS levels. p53 activity is critical for the activation of antioxidant defense mechanisms, which rely on magnesium superoxide dismutase (MnSOD) activity. Failure to activate the p53-MnSOD axis compromises both the acquisition and the maintenance of the HSC phenotype. These studies indicate that suppression of ROS through the coordination of p53 activity and mitochondrial remodeling determines the fate of ex-vivo expanded human HSCs

ORGANISM(S): Homo sapiens

PROVIDER: GSE110968 | GEO | 2018/10/10

REPOSITORIES: GEO

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Ex-vivo Human Hematopoietic Stem Cell Expansion Requires Coordination of Cellular Reprogramming with Mitochondrial Remodeling and P53 Activation[10x]

Project description:Ex-vivo culture conditions used to expand the numbers of hematopoietic stem cells (HSCs) present within an umbilical cord blood (UCB) unit create cellular stresses leading to loss or at best maintenance of the primitive HSCs. Recently, we have shown that treatment with a histone deacetylase inhibitor, valproic acid (VPA), increases substantially the numbers of transplantable HSCs from UCB-CD34+ cells. In this report, we demonstrate that VPA treatment orchestrates cellular mechanisms that drive UCB-CD34+ cells into a primitive state in which they acquire and retain phenotypic, transcriptomic and primitive mitochondrial profiles, all of which characterize long-term HSCs. Remarkably, our data link the acquisition of the HSC phenotype to the remodeling of the mitochondrial network and p53 activation and establish both as critical regulators of ROS and therefore of HSC fate. VPA treatment leads to restructured mitochondria with reduced mass, membrane potential and ROS levels. p53 activity is critical for the activation of antioxidant defense mechanisms, which rely on magnesium superoxide dismutase (MnSOD) activity. Failure to activate the p53-MnSOD axis compromises both the acquisition and the maintenance of the HSC phenotype. These studies indicate that suppression of ROS through the coordination of p53 activity and mitochondrial remodeling determines the fate of ex-vivo expanded human HSCs

2018-10-10 | GSE110973 | GEO

Prostaglandin dose response on hematopoietic stem cells (25 & 37 deg C)

Project description:Umbilical cord blood (UCB) is a valuable source of hematopoietic stem cells (HSCs) for use in allogeneic transplantation. Key advantages of UCB are rapid availability and less stringent requirements for HLA matching. However, UCB contains an inherently limited HSC count, which is associated with delayed time to engraftment, high graft failure rates and early mortality. 16,16 dimethyl prostaglandin E2 (dmPGE2) was previously identified to be a critical regulator of HSC homeostasis and we hypothesized that a brief ex vivo modulation could improve patient outcomes by increasing the "effective dose" of HSCs. Molecular profiling with Affymetrix GeneChips were used to evaluate if increasing the concentration of prostaglandin at 25 and 37 deg C could increase biological responses. Isolated human CD34+ cells from umbilical cord blood were incubated ex vivo in Stem Span (SS) media evaluating five concentrations of 16,16-dmPGE2 (0.1uM, 1uM, 10uM, 50uM, and 100uM) or Vehicle (DMSO) for 2 hours at 25 or 37 deg C. Total RNA was isolated post incubation and analyzed on Affymetrix microarrays for pathway activation.

2013-09-12 | E-GEOD-48541 | biostudies-arrayexpress

CD34+ HSPC CITE-seq for day 0 and Day 1 and Day 4 under VPA

Project description:The regenerative potential of human hematopoietic stem cells (HSCs) is well established by to their ability of life-long blood cell production and cure of a wide range of hematological diseases upon transplantation. This regenerative potential depends on HSC self-renewal and the coordinated adaptation to metabolic stress conditions. This is especially critical during ex vivo culture/manipulation of HSCs that is frequently accompanied with loss of self-renewal potential resulting in stem cell exhaustion. We have previously reported that CD34+ human hematopoietic stem and progenitor cells (HSPC) can be efficiently reprogrammed and expanded to phenotypic HSCs with long-term repopulation capacity in the presence of cytokines and valproic acid (VPA). Here, we present evidence that the SIRT1-SIRT3 axis maintains the mitochondrial activity below a critical threshold by coordinating and retaining a transcriptional and metabolic landscape of human HSCs with long-term self-renewal properties during ex vivo HSC reprogramming.

2023-04-11 | GSE218359 | GEO

Prostaglandin-modulated umbilical cord blood hematopoietic stem cells

Project description:Umbilical cord blood (UCB) is a valuable source of hematopoietic stem cells (HSCs) for use in allogeneic transplantation. Key advantages of UCB are rapid availability and less stringent requirements for HLA matching. However, UCB contains an inherently limited HSC count, which is associated with delayed time to engraftment, high graft failure rates and early mortality. 16,16 dimethyl prostaglandin E2 (dmPGE2) was previously identified to be a critical regulator of HSC homeostasis and we hypothesized that a brief ex vivo modulation could improve patient outcomes by increasing the “effective dose” of HSCs. Molecular profiling with Affymetrix GeneChips were used to determine the optimal ex vivo modulation conditions (e.g., temperature and media) for use in a clinical setting by measured pathway induced expression changes. Isolated human CD34+ from umbilical cord blood were incubated ex vivo in Stem Span (SS) media evaluating three treatment temperatures (4 deg C, 25 deg C, and 37 deg C) with 10uM 16,16-dmPGE2 or Vehicle (DMSO) for 2 hours. To evaluate optimal media, similar CD34+ cells were incubated ex vivo in either Stem Span-SFEM (SS) media or 8% Low Molecular Weigh Dextran 40/5% HSA solution (LMD/HSA) with 10uM 16,16-dmPGE2 or Vehicle (DMSO) for 2 hours at 37 deg C. Total RNA was isolated post incubation and analyzed on Affymetrix microarrays for pathway activation.

2013-09-12 | E-GEOD-46569 | biostudies-arrayexpress

Prostaglandin dose response on hematopoietic stem cells (4 deg C)

Project description:Umbilical cord blood (UCB) is a valuable source of hematopoietic stem cells (HSCs) for use in allogeneic transplantation. Key advantages of UCB are rapid availability and less stringent requirements for HLA matching. However, UCB contains an inherently limited HSC count, which is associated with delayed time to engraftment, high graft failure rates and early mortality. 16,16 dimethyl prostaglandin E2 (dmPGE2) was previously identified to be a critical regulator of HSC homeostasis and we hypothesized that a brief ex vivo modulation could improve patient outcomes by increasing the “effective dose” of HSCs. Molecular profiling with Affymetrix GeneChips were used to evaluate if increasing the concentration of prostaglandin could compensate for the reduced biological responses observed with incubations at 4 deg C. Isolated human CD34+ from umbilical cord blood were incubated ex vivo in Stem Span (SS) media evaluating three concentrations of 16,16-dmPGE2 (10uM, 50uM, and 100uM) or Vehicle (DMSO) for 2 hours at 4 deg C. To evaluate if increasing the concentration of prostaglandin could compensate for the reduced biological responses observed with incubations at 4 deg C. Total RNA was isolated post incubation and analyzed on Affymetrix microarrays for pathway activation.

2013-09-12 | E-GEOD-46634 | biostudies-arrayexpress

Prostaglandin duration required to elicit maximum response on hematopoietic stem cells

Project description:Umbilical cord blood (UCB) is a valuable source of hematopoietic stem cells (HSCs) for use in allogeneic transplantation. Key advantages of UCB are rapid availability and less stringent requirements for HLA matching. However, UCB contains an inherently limited HSC count, which is associated with delayed time to engraftment, high graft failure rates and early mortality. 16,16 dimethyl prostaglandin E2 (dmPGE2) was previously identified to be a critical regulator of HSC homeostasis and we hypothesized that a brief ex vivo modulation could improve patient outcomes by increasing the “effective dose” of HSCs. Molecular profiling with Affymetrix GeneChips were used to evaluate if prostgandin is required for the entire 2 hour incubation to elicit the maximum pathway activated gene expression response. Isolated human CD34+ from umbilical cord blood were incubated ex vivo in Stem Span (SS) media with 10uM 16,16-dimethyl prostaglandin E2 for varying amounts of time within a two hour incubation window to evaluate if the entire 120 minutes is required to elicit the maximum pathway activated gene expression response or if shorter incubation times were sufficent. Total RNA was isolated post incubation and analyzed on Affymetrix microarrays for pathway activation.

2013-09-12 | E-GEOD-46714 | biostudies-arrayexpress

Comparative analysis of gene expression by human Umblical Cord (UCB)-CD34+ Hematopoietic Stem Cells (HSCs) and human induced pluripotent stem (iPS) cell-derived CD34+ Hematopoietic Progenitor Cells (HPCs)

Project description:Hematopoietic stem and progenitor cells are a rare, self-renewing bone marrow resident population capable of giving rise to all circulating hematopoietic cells. They can be used therapuetically for reconstituting defective or ablated hematopoietic systems following chemotherapy, and for inducing tolerance toward allografts of the same haplotype as the HSC donor. There are several sources for HSCs, such as the adult bone marrow, or umblical cord blood, which is more replete with such HSCs. However, HSCs obtained from such sources may be immunogenic, especially if isolated from adult bone marrow. To overcome this issue, our lab has establsihed human induced pluripotent stem cell-derived HPCs with the hope of creating a nonimmunogenic, readily available and unlimited source of HSCs to use for therapy. The goal of this study was to compare the gene expression profiles of naturally found HSCs (UCB-CD34+ HSCs) and HPCs differentiated from 4 different human iPS cell lines (iPS-HPCs), so as to determine the variation between the four iPS-HPCs and whether there were any differences between these HPCs and naturally found HSCs. We utilized 4 iPS cells for this study (detailed descriptions are provided below). iPS cells were differentiated into hematopoietic progenitor cells by coculture on OP9 stromal cells, followed by enrichment of CD34+ cells through immunomagnetic bead separation. The UCB-CD34+ cells were isolated from frozen cord samples through immunomagnetic bead separation. Total RNA was isolated and human gene Affymetrix ST 1.0 arrays performed at the University of Iowa DNA core facility. Data was analyzed, normalized and plotted on BRB Array Tools.

2013-10-23 | E-GEOD-46888 | biostudies-arrayexpress

Re-specification of myeloid precursors from human pluripotent cells into engraftable multi-lineage progenitors

Project description:Human pluripotent stem cells were differentiated into hematopoietic progenitors, which were then re-specified using defined transcription factors to resemble hematopoietic stem cells (HSC) We used microarrays to establish the similarity between converted cells and purified human HSCs. The samples analyzed were: starting embryoid body progenitors, transcription factor-converted cells, and primary HSCs and progenitors from fetal liver and cord blood. All samples were flow sorted for CD34+ and CD38- to compare across a similar population of primitive cells.

2013-11-01 | E-GEOD-49938 | biostudies-arrayexpress

Nynrin preserves hematopoietic stem cell function by inhibiting the mitochondrial permeability transition pore opening （human_HSC_RNA_Seq)

Project description:Mitochondria have recently been identified as a critical regulator for the homeostasis of hematopoietic stem cells (HSCs). However, the mechanism underlying HSC regulation still needs to be clarified. Here, we identify transcription factor Nynrin as a novel regulator of HSC maintenance through modulation of mitochondrial function. We demonstrate that Nynrin is highly expressed in HSC under steady and stress state. Nynrin knockout leads to significantly decreased long-term HSC frequency, markedly reduced HSC dormancy, and self-renewal capacity in steady-state and stress hematopoiesis. We observed abnormal mitochondrial metabolism and mitochondrial permeability transition pore (mPTP) opening in Nynrin-deficient HSCs. Notably, Nynrin-deficient HSCs are more compromised in tolerance of irradiation- and 5-fluorouracil-induced stresses and exhibit typical phenotypes of necrosis. In contrast, overexpression of Nynrin in HSC is resistant to the radiation. Mechanistically, Nynrin deletion induces transactivation of Ppif. Overexpression of cyclophilin D (CypD), the protein encoded by the Ppif gene, causes mPTP opening, mitochondrial swelling, ROS overinduction, and cell necrosis. Both blocking the function of CypD by using cyclosporin A (CsA) or reducing the expression of Ppif could inhibit Nynrin deficiency-induced mitochondrial metabolism enhancement and ROS overproduction, thereby evidently rescuing the impairment of HSCs in Nynrin mutant mice. Collectively, our data, for the first time, characterize Nynrin as a critical regulator of HSC function acting through the Ppif/mPTP mitochondria axis and highlight the importance of Nynrin in HSC maintenance. These data provide new insights into the mechanisms for controlling HSC fate.

2024-06-01 | GSE248636 | GEO