scholarly journals Use of Differentiating Adult Stem Cells (Marrow Stromal Cells) to Identify New Downstream Target Genes for Transcription Factors

Stem Cells ◽  
2006 ◽  
Vol 24 (3) ◽  
pp. 642-652 ◽  
Author(s):  
Joni Ylöstalo ◽  
Jason R. Smith ◽  
Radhika R. Pochampally ◽  
Robert Matz ◽  
Ichiro Sekiya ◽  
...  
Keyword(s):  
Stem Cells ◽  
Transcription Factors ◽  
Stromal Cells ◽  
Adult Stem Cells ◽  
Target Genes ◽  
Marrow Stromal Cells ◽  
Downstream Target

A subset of human rapidly self-renewing marrow stromal cells preferentially engraft in mice

Blood ◽  
2006 ◽  
Vol 107 (5) ◽  
pp. 2153-2161 ◽  
Author(s):  
Ryang Hwa Lee ◽  
Shu Ching Hsu ◽  
James Munoz ◽  
Jin Sup Jung ◽  
Na Rea Lee ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Progenitor Cells ◽  
Stromal Cells ◽  
Adult Stem Cells ◽  
Tissue Injury ◽  
Marrow Stromal Cells ◽  
Immunodeficient Mice ◽  
Adult Mice

Controversies have arisen as to whether adult stem cells or progenitor cells from bone marrow can engraft into nonhematopoietic tissues in vivo. To resolve some of the controversies, we developed a highly sensitive polymerase chain reaction-based single nucleotide polymorphism (PCR-SNP) assay for competitive engraftment of mixtures of stem/progenitor cells. We used the assay to follow engraftment in immunodeficient mice of subpopulations of the stem/progenitor cells from human bone marrow referred to as either mesenchymal stem cells or marrow stromal cells (MSCs). The engraftment into adult mice without induced tissue injury was low and variable, but there was preferential engraftment of a subpopulation of rapidly self-renewing MSCs (RS-MSCs) compared with a subpopulation of slowly renewing MSCs (SR-MSCs). After intravenous infusion, there was a tendency for the cells to engraft into the hippocampal region that was previously designated a “vascular niche.” Migration assays suggested that preferential engraftment of RS-MSCs was in part explained by their expression of CXCR4 and CX3R1, the receptors for SDF-1 and fractalkine.

Transcription Factors Analysis in Neuronal-Like Cells Derived from Adult Human Bone Marrow.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4172-4172
Author(s):  
Shlomo Bulvik ◽  
Tali Ben-Zur ◽  
Yossef S. Levy ◽  
Alex Burshtein ◽  
Eldad Melamed ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Transcription Factors ◽  
Cell Therapy ◽  
Neurodegenerative Diseases ◽  
Stromal Cells ◽  
Bone Marrow Stromal Cells ◽  
Marrow Stromal Cells ◽  
Human Bone ◽  
Human Bone Marrow

Abstract Stem cells are immature cells with self-renewal capacity and ability to differentiate into multiple cell types including bone, cartilage, fat, tendon and muscle. Transplantation of stem cells or their derivatives to the adult brain have been proposed as future therapy for neurodegenerative diseases. Parkinson’s disease (PD) is a progressive neurodegenerative disease involving the deterioration of dopaminergic neurons of the substantia nigra, a part of the midbrain. Most of the currently used drugs are based on dopamine replacement which, although it reduces symptoms is limited owing to severe side effects. Alternatively, strategies of cell therapy for the treatment of PD focused on replacing damaged neurons with cells that may improve the functioning of the damaged cell population. In this study, we demonstrate that bone marrow stromal cells isolated from human iliac crest can be induced to differentiate into neural-like cells. Human bone marrow stromal cells differentiation was induced with N2 supplements, retinoic acid, butylated hydroxyanisole, isobutylmethylxanthine and dibutyryl-cyclic-AMP. A few hours after differentiation the cells changed their phenotype to neural-like cells, spindle shaped, with cell bodies and long branches. Moreover, immunostaining demonstrated the expression of specific neuronal markers such as nestin, neurofilaments and neuron specific enolase. To define the transcription factors associated with the differentiation, we used the TranSignal Protein/DNA kit (Panomics Inc). This new array-based technology allows functional analysis of dozens of eukaryotic transcription factors. Nuclear proteins were purified and incubated with mix labeled DNA binding oligonucleotides corresponding to the consensus sequences of 96 transcription factors. The DNA probes were then dissociated from the complexes and used to hybridize the TranSignal Array. We found that the differentiated human bone marrow stromal cells induced the expression of several neuron-specific transcription factors such as GAG (glycosaminoglycan), FKHR (human forkhead in rhabdomyosarcoma) and DR-2. The activation of these transcription factors in a time course manner was evident also by electrophoretic mobility shift assay. In conclusion, we show that the induced differentiation of neuronal like cells from bone marrow is associated with activation of specific neuronal transcription factors. Our study advances the possible use of autologous transplantation as a new strategy for cell therapy in neurodegenerative diseases.

Serum deprivation of human marrow stromal cells (hMSCs) selects for a subpopulation of early progenitor cells with enhanced expression of OCT-4 and other embryonic genes

Blood ◽  
2004 ◽  
Vol 103 (5) ◽  
pp. 1647-1652 ◽  
Author(s):  
Radhika R. Pochampally ◽  
Jason R. Smith ◽  
Joni Ylostalo ◽  
Darwin J. Prockop
Keyword(s):  
Stem Cells ◽  
Progenitor Cells ◽  
Stromal Cells ◽  
Adult Stem Cells ◽  
Serum Deprivation ◽  
Marrow Stromal Cells ◽  
Embryonic Cells ◽  
Early Passage ◽  
Gene Therapies ◽  
Enhanced Expression

AbstractRecently there has been interest in developing cell and gene therapies with adult stem cells from human bone marrow referred to as mesenchymal stem cells or marrow stromal cells (hMSCs). We incubated early-passage hMSCs in serum-free medium without cytokines or other supplements for 2 to 4 weeks. Surprisingly, a subpopulation of the cells survived serum deprivation and then began to proliferate in serum-containing medium. The cells selected by serum deprivation had longer telomeres than control cells. Also, the patterns of gene expression revealed by reverse transcriptase–polymerase chain reaction (RT-PCR) assays and microarray data indicated that the cells selected by serum deprivation were a subpopulation of very early progenitor cells with enhanced expression of octomer-binding transcription factor 4 (OCT-4) and several other genes characteristically expressed in embryonic cells.

scholarly journals HDAC Inhibition by SNDX-275 (Entinostat) Restores Expression of Silenced Leukemia-Associated Transcription Factors Nur77 and Nor1 and of Key Pro-Apoptotic Proteins in AML

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 311-311 ◽  
Author(s):  
Liran Zhou ◽  
Marina Konopleva ◽  
Michael Andreeff
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Cell Death ◽  
Transcription Factors ◽  
Cell Lines ◽  
Nuclear Receptors ◽  
Target Genes ◽  
Leukemia Stem Cells ◽  
Hdac Inhibition ◽  
Downstream Target

Abstract Orphan nuclear receptors Nur77 & Nor1 are highly conserved members of the nuclear receptor family 4. They are involved in various physiological and cellular functions. Both proteins are products of immediate early genes triggered by growth factors or chemokines. Induction and activation of Nor-1 and/or Nur77 have been implicated in apoptosis for negative selection of T-lymphocytes and activation induced cell death of macrophages. In a previous study (Nat Med. 2007 Jun; 13(6):730–5) we reported that double knockout of Nur77 and Nor1 in mice resulted in the rapid development of AML. The disease was characterized with quick death of mice (2–4 weeks), abnormal expansion of hematopoietic stem cells (HSCs) and myeloid progenitors, hepatosplenomegaly, and lymphadenopathy. Molecular analysis revealed that the absence of both Nur77 and Nor1 leads to the downregulation of AP1 transcription factors c-Jun and JunB, and extrinsic apoptosis mediators TRAIL and FasL. Real- time RT-PCR analysis of AML patient samples (n=16) revealed the universal down-regulation of Nur77 and Nor1 transcripts in AML progenitor and stem cells (CD34+38−). These findings indicate that Nur77 and Nor1 play roles in maintaining hematopoiesis and silencing of Nur77 and Nor1 is a critical step in AML development. To investigate the role of Nur77/Nor1 in AML, we hypothesized that silenced gene expression of Nur7 and Nor1 is mediated by histone deacetylases (HDACs). HDACs are enzymes involved in chromatin modifications that play an important role in the epigenetic regulation of gene expression and mediate abnormal epigenetic alterations associated with cancer, which make HDAC inhibitors promising anti-cancer drugs. SNDX-275, a benzamide HDAC inhibitor that preferentially inhibits activities of nuclear HDACs, including HDAC1, HDAC2, and HDAC3, has been shown to induce extensive apoptosis in various cancer types and has progressed to clinical phase I and II trials. We demonstrated that HDAC inhibition with SNDX-275 restored the expression of Nur77 and Nor1 in AML cell lines and primary AML samples (n=5) to levels comparable to those found in normal bone marrows. In the knockout mouse model, TRAIL and AP-1 transcription factors (c-Jun and JunB) were identified as downstream target genes of Nur77 and Nor1. In our present study, we observed strong upregulation by SNDX-275 of TRAIL, c-Jun and JunB in leukemia cell lines and primary AML blasts. As is common with HDAC inihibitors, apoptosis induced by SNDX-275 was associated with dramatic H3 and H4 acetylation. We further exploited the lack of Nur77 and Nor1 in leukemic stem cells (CD34+/38−) and demonstrated that SNDX-275 restored their expression resulting in apoptotic cell death of leukemia stem cells. Besides, pro-apoptotic Bim and Noxa were transcriptionally upregulated by SNDX-275 in AML cell lines, primary AML samples and also leukemia stem cells. Altogether, our results suggest a novel mechanism of HDAC inhibitor-induced apoptosis in AML which involves the restoration of the silenced nuclear receptors Nur77 and Nor1 and the activation of their downstream target genes, in addition to affecting apoptotic machinery. These findings also characterize HDAC-mediated transcriptional silencing of nuclear receptors as one of the key mechanisms driving the pathogenesis of AML.

scholarly journals Rat adult stem cells (marrow stromal cells) engraft and differentiate in chick embryos without evidence of cell fusion

2004 ◽  
Vol 101 (25) ◽  
pp. 9282-9285 ◽  
Author(s):  
R. R. Pochampally ◽  
B. T. Neville ◽  
E. J. Schwarz ◽  
M. M. Li ◽  
D. J. Prockop
Keyword(s):  
Stem Cells ◽  
Cell Fusion ◽  
Stromal Cells ◽  
Adult Stem Cells ◽  
Marrow Stromal Cells ◽  
Chick Embryos

scholarly journals Long non‐coding RNA NORAD promotes the prostate cancer cell extracellular vesicle release via microRNA-541-3p-regulated PKM2 to induce bone metastasis of prostate cancer

2021 ◽  
Vol 40 (1) ◽  
Author(s):  
Chuan-yi Hu ◽  
Juan Chen ◽  
Xin-hua Qin ◽  
Pan You ◽  
Jie Ma ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Bone Marrow ◽  
Bone Metastasis ◽  
Stromal Cells ◽  
Bone Marrow Stromal Cells ◽  
Marrow Stromal Cells ◽  
Downstream Target ◽  
Non Coding Rna ◽  
Target Molecules ◽  
Long Non Coding Rna

Abstract Background Bone metastasis is the leading cause of mortality and reduced quality of life in patients with metastatic prostate cancer (PCa). Long non-coding RNA activated by DNA damage (NORAD) has been observed to have an abnormal expression in various cancers. This article aimed to explore the molecular mechanism underlying the regulatory role of NORAD in bone metastasis of PCa. Methods NORAD expression in clinical PCa tissues and cell lines was detected with the application of qRT-PCR. Cancer cells were then transfected with plasmids expressing NORAD, after which Transwell assay and CCK-8 assay were carried out to detect proliferation, migration, and bone metastasis of PCa. NORAD downstream target molecules were screened through bioinformatics analysis, followed by further verification using dual luciferase assay. Extracellular vesicles (EVs) were labeled with PKH67 and interacted with bone marrow stromal cells. The gain- and loss-function method was applied to determine the internalization and secretion of PCa cells-derived EVs under the intervention of downstream target molecules or NORAD. Results PCa tissues and cell lines were observed to have a high expression of NORAD, particularly in tissues with bone metastasis. NORAD knockdown resulted in reduced secretion and internalization of EVs, and suppressed proliferation, migration, and bone metastasis of PCa cells. It was indicated that NORAD interacted with miR-541-3p, leading to the upregulation of PKM2. Forced expression of PKM2 promoted the transfer of PKH67-labeled EVs to bone marrow stromal cells. Conclusions NORAD might serve as a ceRNA of miR-541-3p to promote PKM2 expression, thereby enhancing the development of bone metastasis in PCa by promoting internalization and transfer of EVs of cancer cells, providing an insight into a novel treatment for the disorder.

scholarly journals Biological and microbiological interactions of Ti-35Nb-7Zr alloy and its basic elements on bone marrow stromal cells: good prospects for bone tissue engineering

2020 ◽  
Vol 6 (1) ◽  
Author(s):  
Daphne de Camargo Reis Mello ◽  
Lais Morandini Rodrigues ◽  
Fabia Zampieri D’Antola Mello ◽  
Thais Fernanda Gonçalves ◽  
Bento Ferreira ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Cell Differentiation ◽  
Stromal Cells ◽  
Biofilm Formation ◽  
Bone Marrow Stromal Cells ◽  
Pure Titanium ◽  
Marrow Stromal Cells ◽  
The Other ◽  
Cell Based Therapy

Abstract Background An effective biomaterial for bone replacement should have properties to avoid bacterial contamination and promote bone formation while inducing rapid cell differentiation simultaneously. Bone marrow stem cells are currently being investigated because of their known potential for differentiation in osteoblast lineage. This makes these cells a good option for stem cell-based therapy. We have aimed to analyze, in vitro, the potential of pure titanium (Ti), Ti-35Nb-7Zr alloy (A), niobium (Nb), and zirconia (Zr) to avoid the microorganisms S. aureus (S.a) and P. aeruginosa (P.a). Furthermore, our objective was to evaluate if the basic elements of Ti-35Nb-7Zr alloy have any influence on bone marrow stromal cells, the source of stem cells, and observe if these metals have properties to induce cell differentiation into osteoblasts. Methods Bone marrow stromal cells (BMSC) were obtained from mice femurs and cultured in osteogenic media without dexamethasone as an external source of cell differentiation. The samples were divided into Ti-35Nb-7Zr alloy (A), pure titanium (Ti), Nb (niobium), and Zr (zirconia) and were characterized by scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS). After predetermined periods, cell interaction, cytotoxicity, proliferation, and cell differentiation tests were performed. For monotypic biofilm formation, standardized suspensions (106 cells/ml) with the microorganisms S. aureus (S.a) and P. aeruginosa (P.a) were cultured for 24 h on the samples and submitted to an MTT test. Results All samples presented cell proliferation, growth, and spreading. All groups presented cell viability above 70%, but the alloy (A) showed better results, with statistical differences from Nb and Zr samples. Zr expressed higher ALP activity and was statistically different from the other groups (p < 0.05). In contrast, no statistical difference was observed between the samples as regards mineralization nodules. Lower biofilm formation of S.a and P.a. was observed on the Nb samples, with statistical differences from the other samples. Conclusion Our results suggest that the basic elements present in the alloy have osteoinductive characteristics, and Zr has a good influence on bone marrow stromal cell differentiation. We also believe that Nb has the best potential for reducing the formation of microbial biofilms.

Cardiac Transcription Factors Driven Lineage-Specification of Adult Stem Cells

2009 ◽  
Vol 3 (1) ◽  
pp. 61-65 ◽  
Author(s):  
Ana Armiñán ◽  
Carolina Gandía ◽  
José Manuel García-Verdugo ◽  
Elisa Lledó ◽  
José Luis Mullor ◽  
...  
Keyword(s):  
Stem Cells ◽  
Transcription Factors ◽  
Adult Stem Cells ◽  
Lineage Specification ◽  
Cardiac Transcription Factors
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