Hypoxia Drives AML Proliferation in the Bone Marrow Microenvironment Via Macrophage Inhibitory Factor

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 1721-1721
Author(s):  
Amina M Abdul-Aziz ◽  
Manar S Shafat ◽  
Lyubov Zaitseva ◽  
Matthew J Lawes ◽  
Stuart A Rushworth ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cell Proliferation ◽  
Protein Expression ◽  
Mrna Expression ◽  
Cell Lines ◽  
Bone Marrow Microenvironment ◽  
Inhibitory Factor ◽  
Bone Marrow Niche ◽  
Hematopoietic Stem ◽  
Hypoxic Conditions

Abstract Introduction Hypoxia is an important component of the bone marrow microenvironment and the hematopoietic stem cell niche. Studies have shown that hypoxia contributes to the development and maintenance of acute myeloid leukemia (AML) cells within the bone marrow microenvironment. Hypoxia is principally maintained by members of the hypoxia-inducible factor (HIF), in particular HIF1a and its target genes, including MIF. We have previously shown that AML cells express constitutively high macrophage migration inhibitory factor (MIF) which drives IL-8 expression by the BM-MSC which in turn supports AML cell survival and proliferation. The aim of the present study is to determine if there is a connection between the role of hypoxia in regulating AML survival and MIF survival signals. Furthermore we investigate the role BM-MSC in regulating the hypoxic response. Methods Primary AML and BM-MSC were isolated from AML patients following informed consent and under approval from the UK National Research Ethics Service (LRCEref07/H0310/146). AML cell lines and primary AML blasts were cultured under normoxic (20% oxygen) or hypoxic conditions (1% oxygen) for 4 - 24 hours, mRNA expression of MIF, HIF1a, VEGF and IL-8 were determined by RT-PCR. MIF and IL-8 protein was determined using target specific ELISA. HIF1a protein expression was determined by western blotting. Hypoxia-mimetic agents, cobalt chloride (CoCl2) and desferrioxamine (DFO) were used. Cell proliferation was determines using CellTiter Glo and trypan blue exclusion. CFU-assays were performed using complete methylcellulose media. Results To determine If MIF is regulated by HIF1a in AML cells, we mimicked hypoxic conditions using CoCl2 and DFO in AML cells. Both CoCl2 and DFO upregulate MIF transcription and protein expression in OCI-AML3 cell lines and in primary AML blasts. Moreover, hypoxia increases both MIF mRNA expression and MIF chemokine expression compared to normoxic conditions. Lentiviral mediated knockdown of MIF in AML cells show significantly reduced cell proliferation and colony formation in methylcellulose media. Recombinant MIF induced interleukin-8 in AML blasts and the MIF inhibitor blocked MIF induced IL-8 release. Lentiviral mediated KD of HIF1a decreased MIF expression in human AML cells and a significant reduced their proliferative capacity. Finally we found that hypoxia increased MIF in AML blasts which was further increased when in co-culture with BM-MSC. Conclusions The results reported here suggest that hypoxia significantly affects the expression of the survival cytokine MIF in AML blasts. Furthermore, we show that BM-MSC regulate HIF1a induced MIF expression in AML under hypoxic conditions. We propose this hypoxia regulated HIF1a/MIF axis is essential to blast survival in the bone marrow niche. Disclosures Rushworth: Infinity Pharmaceuticals: Research Funding.

The Hypoxic Microenvironment in Acute Myelogenous Leukemia: Critical Role of CXCR4 in the Induction of HIF-1α.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 1819-1819
Author(s):  
Michael Andreeff ◽  
Martin Dietrich ◽  
Paul Corn ◽  
Sergej Konoplev ◽  
Marina Konopleva ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cell Lines ◽  
Critical Role ◽  
Mapk Signaling ◽  
Bone Marrow Microenvironment ◽  
Myelogenous Leukemia ◽  
Leukemic Cells ◽  
Mrna Levels ◽  
Hematopoietic Stem ◽  
Hypoxic Conditions

Abstract The bone marrow microenvironment provides functional and structural support for both normal and leukemic hematopoietic stem cells. Importantly, the marrow microenvironment is known to be hypoxic. Given the many critical functions of HIF-1α, we investigated HIF-1α levels in leukemic cell lines and primary AML blasts. Surprisingly, HIF-1α expression was non-detectable in AML cells grown in suspension cultures under normoxic (21% O2) or hypoxic (1% O2) conditions. Normoxic co-cultures with bone marrow-derived mesenchymal stromal (MSC) or MS5 cells likewise did not induce HIF-1α, but hypoxic co-culture conditions induced HIF-1α protein without changes in HIF-1α mRNA levels, suggesting post-transcriptional regulation. Functionality of HIF-1α was confirmed by concomitant increase in the levels of glucose transporter glut-1, the HIF-1α downstream target. Inhibition of stroma-leukemia cell interactions with the small molecule CXCR4 inhibitor AMD3465 (Genzyme/Anormed) at 100 nM completely abrogated the induction of HIF-1α in HL-60 and MOLM13 AML cells. While SDF-1 was unable to induced HIF-1α under normoxic conditions, it did so under both physical (1% O2) and chemical (COCl2) hypoxic conditions, in two different cell lines. Inhibition of P13K (with LY294002) or MEK/ERK signaling (with CI-1040) abrogated HIF-1α induction under hypoxic conditions. Immunohistochemical staining of bone marrow samples from primary AML confirmed the presence of HIF-1α in leukemic cells localized adjacent to bone-lining stromal elements. Results suggest that the bone marrow microenvironment of AML is hypoxic in vivo; in leukemia cells HIF-1α induction under low oxygen tension depends on the presence of stromal cells; HIF-1α induction is dependent on SDF-1/CXCR4 and is mediated by activation of P13K and MAPK signaling. Altogether these findings suggest that SDF-1α/CXCR4 interactions contribute to the survival of leukemic cells via specific induction of HIF-1α signaling by the bone marrow microenvironment. Disruption of these interactions via CXCR4 inhibition strategies may suppress multiple pro-survival HIF-1α targets in leukemic cells.

Mtss1 Suppresses BCR-ABL Induced Cell Migration and Is Downregulated in CML Stem Cells.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 1077-1077
Author(s):  
Mirle Schemionek ◽  
Shuchi Agrawal ◽  
Martin Stehling ◽  
Daniel G. Tenen ◽  
Ashley Hamilton ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Cell Migration ◽  
Tumor Suppressor ◽  
Transgenic Mice ◽  
Cell Lines ◽  
Actin Binding ◽  
Bone Marrow Niche ◽  
Hematopoietic Stem ◽  
Retroviral Transduction

Abstract Migration and adhesion properties of hematopoietic stem cells (HSC) are disrupted in chronic myeloid leukemia (CML). Egression of these cells from the bone marrow is associated with cytoskeletal changes including actin remodeling. In a microarray screen of differentially regulated genes in HSC from BCR-ABL positive transgenic mice, we found downregulation of multiple genes involved in actin-associated changes of cell structure, adhesion, and migration (i.e. intersectin-1, cortactin, Mtss1, synaptopodin, and Gem GTPase). Mtss1 was further studied since it has been described to be a binding partner of Rac, which is essential for BCR-ABL mediated transformation, and a potential tumor suppressor. Using quanitative RT-PCR, Mtss1 downregulation (6-fold) was confirmed in HSC and unfractionated bone marrow and spleen cells from SCLtTA/BCR-ABL transgenic mice after 3 weeks of BCR-ABL induction as well as in human BCR-ABL positive cell lines. Treatment of BCR-ABL positive (32D/BCR-ABL, K562, KYO-1) but not BCR-ABL negative (32D, U937) cell lines with 5μ M Imatinib led to upregulation of Mtss1 mRNA (5- to 10-fold) and protein, suggesting that downregulation of Mtss1 is dependent on BCR-ABL kinase activity. Retroviral transduction of Mtss1 into 32D/BCR-ABL cells almost completely inhibited BCR-ABL induced cell motility of individual cells seeded on murine bone marrow stromal cells in time-lapse video experiments over the course of two hours. Interestingly, we found that retroviral transduction of Mtss1 into 32D/BCR-ABL cells completely suppressed migration of these cells to extra-hematopoietic sites in vivo upon intravenous transplantation into syngeneic C3H mice. Moreover, when Mtss1-transduced cells were injected subcutaneously, the size of the tumors was significantly decreased as compared to empty vector-transduced 32D/BCR-ABL cells (p<0.05), confirming that Mtss1 may be a tumor suppressor. These results demonstrate that Mtss1 antagonizes BCR-ABL induced cell migration and is downregulated by BCR-ABL in CML stem cells, suggesting that downregulation of Mtss1 and other cytoskeletal adaptor proteins may be required for egression of CML stem cells from the bone marrow niche. Since the same Mtss1 protein domain is responsible for both Rac and actin binding, Mtss1 may interfere with Rac function and thereby inhibit the effects of Rac on migration and cytoskeletal dynamics of HSC.

scholarly journals Alterations of Bone Marrow Microenvironment in Acute Graft-Versus-Host Disease

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 5009-5009
Author(s):  
Yan Lin ◽  
Quan Gu ◽  
Hui Cheng ◽  
Zhaofeng Zheng ◽  
Sun Guohuan ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stem Cell ◽  
Graft Versus Host Disease ◽  
Ppar Gamma ◽  
Bone Marrow Microenvironment ◽  
Bone Marrow Niche ◽  
Hematopoietic Stem ◽  
Host Disease ◽  
Graft Versus Host ◽  
Acute Graft

Severe acute graft-versus-host disease (aGVHD) indicates a poor prognosis after allogeneic hematopoietic stem cell transplantation. In our previous study, we found that hematopoiesis was progressively suppressed during aGVHD, while the hematopoietic regenerative potential of donor-derived hematopoietic stem cells remains intact. It prompts us to investigate whether bone marrow niche is a major target of GVHD. We addressed this issue by studying the critical components in bone marrow microenvironment, including mesenchymal stem cell (MSC), osteoblasts and adipocytes in haplo-MHC-matched murine bone marrow transplantation model. By comparing confocal images of femurs from control and aGVHD SCF-GFP mice or Col2.3-GFP mice, we found that both MSCs and osteoblasts were significantly reduced during aGVHD development. In addition, anti-perilipin staining showed that adipocytes were also decreased in aGVHD mice. We found a defect in the differentiation potential of MSCs from aGVHD niche by in vitro culture of both murine and human bone marrow niche cells. qRT-PCR showed decreased gene expressions of PPAR-gamma, Adipoq, Runx1 and Col2a1, suggesting the potential of MSCs differentiation into adipocytes and osteoblasts was blocked. These data provide new insights into the pathogenesis of aGVHD and may improve the clinical management of aGVHD. Disclosures No relevant conflicts of interest to declare.

scholarly journals The IL1-IL1RAP axis plays an important role in the inflammatory leukemic niche that favors acute myeloid leukemia proliferation over normal hematopoiesis.

Haematologica ◽  
2020 ◽  
pp. 0-0
Author(s):  
Bauke De Boer ◽  
Sofia Sheveleva ◽  
Katja Apelt ◽  
Edo Vellenga ◽  
Andre B. Mulder ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cell Proliferation ◽  
Acute Myeloid Leukemia ◽  
Myeloid Leukemia ◽  
Bone Marrow Niche ◽  
Hematopoietic Stem ◽  
Signaling Axis ◽  
Plasma Membrane Receptor ◽  
Colony Forming Capacity ◽  
Acute Myeloid

Upregulation of the plasma membrane receptor IL1RAP in Acute Myeloid Leukemia (AML) has been reported but its role in the context of the leukemic bone marrow niche is unclear. Here, we studied the signaling events downstream of IL1RAP in relation to leukemogenesis and normal hematopoiesis. High IL1RAP expression was associated with a leukemic GMP-like state, and knockdown of IL1RAP in AML reduced colony-forming capacity. Stimulation with IL1β resulted in the induction of multiple chemokines and an inflammatory secretome via the p38 MAPK and NFκB signaling pathways in IL1RAP-expressing AML cells, but IL1β-induced signaling was dispensable for AML cell proliferation and NFκB-driven survival. IL1RAP was also expressed in stromal cells where IL1β induced expression of inflammatory chemokines and cytokines as well. Intriguingly, the IL1β-induced inflammatory secretome of IL1RAPexpressing AML cells grown on a stromal layer of mesenchymal stem cells affected normal hematopoiesis including hematopoietic stem/progenitor cells while AML cell proliferation was not affected. The addition of Anakinra, an FDA-approved IL1 receptor antagonist, could reverse this effect. Therefore, blocking the IL1-IL1RAP signaling axis might be a good therapeutic approach to reduce inflammation in the bone marrow niche and thereby promote normal hematopoietic recovery over AML proliferation after chemotherapy.

High Mobility Group A1 (HMGA1) Chromatin Remodeling Protein Mediates Crosstalk Between Acute Myeloid Leukemia Blasts & the Tumor Microenvironment

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 3564-3564 ◽  
Author(s):  
Stuart A Rushworth ◽  
Lyubov Zaitseva ◽  
Lingling Xian ◽  
Kristian M Bowles ◽  
Linda M. S. Resar
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Cell Lines ◽  
Chromatin Remodeling ◽  
High Mobility ◽  
Bone Marrow Microenvironment ◽  
High Mobility Group ◽  
Hematopoietic Stem ◽  
Knock Down ◽  
Acute Myeloid

Abstract Introduction: Acute myeloid leukemias (AMLs) are highly lethal hematologic malignancies that arise from diverse genetic abnormalities in hematopoietic progenitor cells. Unfortunately, most patients with AML will die of their disease due to failure of currently available cytotoxic chemotherapies. Accordingly, there is an urgent need to better understand the biologic interactions between AML blasts and signals from bone marrow stromal cells (BMSCs) within the niche that support their survival and protect them from exposure to chemotherapy. The high mobility group A1 (HMGA1) chromatin remodeling proteins are present at high levels during development and enriched in AML blasts, leukemic stem cells, diverse solid tumors, embryonic stem cells, and adult stem cells, such as hematopoietic stem cells (HSCs). HMGA1 proteins regulate gene expression by modulating chromatin structure and recruiting NF-κB and other transcription factor complexes to DNA. We discovered that HMGA1 acts as a potent oncogene in transgenic mouse and cultured cell models (Xu et. al, Cancer Research, 2004) by inducing stem cell transcriptional networks (Schuldenfrei et. al, BMC Genomics, 2011, Shah et. al, PLoS ONE, 2013). Here, we describe a novel role for HMGA1 in mediating crosstalk between AML blasts and BMSCs within the bone marrow microenvironment. Methods & Results: To investigate HMGA1 in regulating AML-niche signaling, we used potent lentiviruses to deliver short hairpin RNA and silence HMGA1 in AML blasts or BMSCs. We found that silencing HMGA1 rapidly halts proliferation and induces apoptotic cell death in 3 different AML cell lines. To determine how HMGA1 mediates survival in AML blasts, we assessed expression of pro-survival genes in AML cell lines and primary AML blasts, including those encoding NF-E2-related factor 2 (NRF2), cMYC, and the C-X-C chemokine receptor type 4 (CXCR4). CXCR4 is the receptor for stromal cell-derived factor 1 (SDF-1 or CXCL12), a growth factor secreted by BMSCs that also serves as a chemo-attractant for AML blasts or HSCs within the bone marrow microenvironment. We found that silencing HMGA1 represses expression of NRF2, cMYC, and CXCR4. This led us to hypothesize that HMGA1 regulates crosstalk between AML blasts and the leukemic cell niche via CXCR4 and SDF-1. To test this, we silenced HMGA1 in cultured AML cells and assessed migration in the presence of SDF-1. Strikingly, migration was significantly impaired in the AML cells with HMGA1 knock-down. Next, we silenced HMGA1 in primary, patient-derived BMSCs, which were co-cultured with primary AML blasts from the same patients to mimic the bone marrow microenvironment. We found that knock-down of HMGA1 in BMSCs also results in apoptosis in primary AML blasts. Conclusions: Together, our results demonstrate for the first time that HMGA1 mediates AML survival through cell-autonomous pathways in AML blasts and through non-cell-autonomous crosstalk from BMSCs within the bone marrow microenvironment. Studies are underway to determine if HMGA1 directly regulates expression of SDF-1 or other factors secreted by BMSCs within the hematopoietic niche. This knowledge should inform biologically rational strategies to enhance existing treatments and facilitate the design of novel therapies. Disclosures No relevant conflicts of interest to declare.

scholarly journals Aging of Bone Marrow Mesenchymal Stromal Cells: Hematopoiesis Disturbances and Potential Role in the Development of Hematologic Cancers

Cancers ◽  
2020 ◽  
Vol 13 (1) ◽  
pp. 68
Author(s):  
Fulvio Massaro ◽  
Florent Corrillon ◽  
Basile Stamatopoulos ◽  
Nathalie Meuleman ◽  
Laurence Lagneaux ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Tumor Progression ◽  
Cancer Progression ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Cell Communication ◽  
Bone Marrow Niche ◽  
Hematopoietic Stem ◽  
Wide Range ◽  
Major Player

Aging of bone marrow is a complex process that is involved in the development of many diseases, including hematologic cancers. The results obtained in this field of research, year after year, underline the important role of cross-talk between hematopoietic stem cells and their close environment. In bone marrow, mesenchymal stromal cells (MSCs) are a major player in cell-to-cell communication, presenting a wide range of functionalities, sometimes opposite, depending on the environmental conditions. Although these cells are actively studied for their therapeutic properties, their role in tumor progression remains unclear. One of the reasons for this is that the aging of MSCs has a direct impact on their behavior and on hematopoiesis. In addition, tumor progression is accompanied by dynamic remodeling of the bone marrow niche that may interfere with MSC functions. The present review presents the main features of MSC senescence in bone marrow and their implications in hematologic cancer progression.

scholarly journals Bone marrow stromal cells from MDS and AML patients show increased adipogenic potential with reduced Delta-like-1 expression

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Marie-Theresa Weickert ◽  
Judith S. Hecker ◽  
Michèle C. Buck ◽  
Christina Schreck ◽  
Jennifer Rivière ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cell Fate ◽  
Stromal Cells ◽  
Adipogenic Differentiation ◽  
Cell Types ◽  
Bone Marrow Niche ◽  
Differentiation Potential ◽  
Hematopoietic Stem ◽  
Bm Niche

AbstractMyelodysplastic syndromes (MDS) and acute myeloid leukemia (AML) are clonal hematopoietic stem cell disorders with a poor prognosis, especially for elderly patients. Increasing evidence suggests that alterations in the non-hematopoietic microenvironment (bone marrow niche) can contribute to or initiate malignant transformation and promote disease progression. One of the key components of the bone marrow (BM) niche are BM stromal cells (BMSC) that give rise to osteoblasts and adipocytes. It has been shown that the balance between these two cell types plays an important role in the regulation of hematopoiesis. However, data on the number of BMSC and the regulation of their differentiation balance in the context of hematopoietic malignancies is scarce. We established a stringent flow cytometric protocol for the prospective isolation of a CD73+ CD105+ CD271+ BMSC subpopulation from uncultivated cryopreserved BM of MDS and AML patients as well as age-matched healthy donors. BMSC from MDS and AML patients showed a strongly reduced frequency of CFU-F (colony forming unit-fibroblast). Moreover, we found an altered phenotype and reduced replating efficiency upon passaging of BMSC from MDS and AML samples. Expression analysis of genes involved in adipo- and osteogenic differentiation as well as Wnt- and Notch-signalling pathways showed significantly reduced levels of DLK1, an early adipogenic cell fate inhibitor in MDS and AML BMSC. Matching this observation, functional analysis showed significantly increased in vitro adipogenic differentiation potential in BMSC from MDS and AML patients. Overall, our data show BMSC with a reduced CFU-F capacity, and an altered molecular and functional profile from MDS and AML patients in culture, indicating an increased adipogenic lineage potential that is likely to provide a disease-promoting microenvironment.

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