CXCL12 and IGF1 select for cancer cell populations with enhanced bone metastasis ability
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
SUBMITTER: Xiang Zhang
PROVIDER: E-GEOD-39102 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:CXCL12 and IGF1 confer on cancer cells survival advantage. Src potentiates cancer cells' reponse to CXCL12 and IGF1 by strengthening AKT activation. Long-term incubation of CXCL12 and IGF1 select for cancer cells with enhanced Src activity and bone metastasis potential.
2012-12-10 | GSE39102 | GEO
Project description:CXCL12 and IGF1 are key secreting molecules produced by cancer-associated fibroblasts in breast cancer. These factors promote the survival of disseminated cancer cells in the bone marrow. To assess the combined responses elicited by CXCL12 and IGF1, we examined the translating transcriptome of cancer cells in response to these two factors by Translating Ribosome Affinity Purification (TRAP)-RNAseq. MDA-MB-231 cells were engineered to express an EGFP-tagged version of ribosomal protein L10a. This allows the retrieval of polysome-associated mRNA by anti-GFP pull down (TRAP) and profiling the translating transcriptome by RNAseq. EGFP-L10a+ cancer cells were serum starved (0.2% serum) for 24 hours, and then treated with CXCL12 (30ng/mL) + IGF1 (10ng/mL) or CXCL12 (300ng/mL) + IGF1 (100ng/mL) for 6hrs. Two biological replicates were profiled for each condition.
2013-11-01 | E-GEOD-43296 | biostudies-arrayexpress
Project description:CXCL12 and IGF1 are key secreting molecules produced by cancer-associated fibroblasts in breast cancer. These factors promote the survival of disseminated cancer cells in the bone marrow. To assess the combined responses elicited by CXCL12 and IGF1, we examined the translating transcriptome of cancer cells in response to these two factors by Translating Ribosome Affinity Purification (TRAP)-RNAseq.
2013-11-01 | GSE43296 | GEO
Project description:Homing and engraftment of hematopoietic stem cells (HSCs) to the bone marrow (BM) involve a complex interplay between chemokines, cytokines, and non-peptide molecules. Extracellular nucleotides and their cognate P2 receptors are emerging as key-factors of inflammation and related chemotactic responses. In this study, we investigated the activity of extracellular adenosine-triphosphate (ATP) and uridine-triphosphate (UTP) on CXCL12-stimulated CD34+ HSC chemotaxis. In vitro, UTP significantly improved HSC migration, inhibited cell membrane CXCR4 down-regulation of migrating CD34+ cells and increased cell adhesion to fibronectin. In vivo, pre-incubation with UTP significantly enhanced the BM homing efficiency of human CD34+ cells in immunodeficient mice. Pertussis toxin blocked CXCL12- and UTP-dependent chemotactic responses, suggesting that G-protein alpha-subunits (Gαi) may provide a converging signal for CXCR4- and P2Y-activated transduction pathways. In addition, gene expression profiling of UTP-treated CD34+ cells and in vitro inhibition assays demonstrated that Rho guanosine 5â-triphosphatases (GTPase) Rac2 and downstream effectors Rho GTPaseâactivated kinases 1 and 2 (ROCK1/2) are involved in UTP-promoted/CXCL12-dependent HSC migration. Our data suggest that UTP may physiologically modulate the migration of HSCs and their homing to the BM, in concert with CXCL12, via the activation of converging signaling pathways between CXCR4 and P2Y receptors, involving Gαi proteins and RhoGTPases. Experiment Overall Design: Highly purified CD34+ cells from 6 healthy donors were seeded at 1000000 cells/ml in serum free medium (EX vivo 15) w/o cytokines and treated with 10 mM UTP, 150ng/ml CXCL12, or 10 mM UTP plus 150ng/ml CXCL12 respectively for 24 hours. As a control, CD34+ untreated cells were maintained in the same culture conditions at the same time.
2008-06-13 | E-GEOD-5168 | biostudies-arrayexpress
Project description:The chemokine CXCL12 and its receptor CXCR4 play important roles in signaling and migration of T-cells, but little is known about the transcriptional events involved in CXCL12-mediated T-cell migration. In this study we performed microarray analysis on CXCL12- treated T-cells, and found that the Wnt family of proteins was significantly upregulated during CXCL12 treatment. Confirmation of these results by real-time PCR and Western analysis indicated that the non-canonical Wnt pathway was specifically upregulated during CXCL12 treatment. In vitro and in vivo knockdown studies confirm that b-catenin (the key mediator of canonical Wnt signaling) is not involved in the CXCL12-mediated migration of T-cells. However, Wnt5A, a non-canonical Wnt protein, increases signaling through the CXCL12/ CXCR4 axis via Protein Kinase C (PKC). Our results demonstrated that CXCL12 required Wnt5A to mediate T-cell migration, and the treatment of T-cells with recombinant Wnt5A sensitized T-cells to CXCL12 induced migration. Additionally, Wnt5A expression was required for the sustained expression of CXCR4, both transcriptionally and translationally. These results could be translated in vivo, using EL4 thymoma metastasis as a model of T-cell migration. Taken together our data indicate, for the first time, that Wnt5A is a critical mediator of the CXCL12/ CXCR4 signaling axis. Experiment Overall Design: Primary T cells were treated with human CXCL12 (Peprotech, Rocky Hill, NJ) at 100 ng/ml per 10 million cells overnight in a humidified incubator at 37ºC with 5% CO2. Control cells were incubated in media only. Cells were harvested and washed with ice cold PBS for 2 times followed by the addition of ice cold TRIzol (Invitrogen, Carlsbad, CA) and frozen at -80ºC overnight. Total RNA was isolated using the RNA isolation kit manufactured by Qiagen (Valencia, CA). The cDNA was prepared from equal amount of RNA using a cDNA preparation kit (Bio-Rad, Hercules, CA) followed by preparation of cRNA according to manufacturerâs instructions (Agilent, Santa Clara, CA). The cRNA was amplified and labeled with either Cy-3 or Cy-5, using the Agilent low-input linear amplification kit, according to manufacturerâs protocols. Labeled cRNA were applied to the Human 44K whole genome oligo array slides (Agilent, Santa Clara, CA). Slides were hybridized in a rotating chamber overnight at 60ºC in 6X SSC. Next day, slides were washed with 0.005% Triton X-102 for 10 minutes, and then in 0.1X SSC, 0.005% Triton X-102 for 5 minutes on ice. Slides were dried using a nitrogen-filled air gun, and scanned using an Agilent scanner. Images were analyzed using the Agilent Feature Extractor Software, Version A.7.5.1 and ratios for each spot were calculated.
2009-01-14 | E-GEOD-12449 | biostudies-arrayexpress
Project description:Total mRNA expression of CXCL12-treated monocytes was compared with control untreated monocytes Human Peripheral blood monocytes from three different healthy donors were isolated by anti-CD14-labeled magnetic microbeads. CD14+ monocytes were cultured in teflon dishes for 1h in RPMI 10% FCS and then, were treated or not with CXCL12 for 6h. Total RNA from each condition was extracted and purified using the RNasey kit (Qiagen). Labelled RNA was used as hybridization probes on human Codelink Whole genome Bioarray. All experimental procedures were performed following manufacturer instructions. Microarrays were scanned with a GenePix 4000B (Axon Instruments) scanner. Scanned images and raw data were processed using the Codelink Expression Software.
2011-01-13 | E-GEOD-24496 | biostudies-arrayexpress
Project description:Defects in cell-cell contacts have been proposed to participate in early steps of tumor metastasis1 but little is known about the signals from the cancer cell niche that influence this process. Here, we show that the lack of adherens junctions caused by genetic loss of the adhesion molecule NECTIN1 in melanoma promotes tumor dissemination specifically under growth factor deprivation. We found that NECTIN1 was deleted in 53% of human melanomas and that its loss was enriched in metastases. NECTIN1 inactivation in zebrafish melanomas stimulated cancer cell spreading in vivo, while in human cell lines, it increased cell migration specifically in response to serum starvation. We further identified IGF1 as the serum component responsible for this effect. Serum withdrawal or IGF1 inhibition induced formation of strong adherens junctions between NECTIN1-wild-type melanoma cells. In contrast, NECTIN1-deficient cells were unable to establish adherens junctions and instead activated integrin-mediated motility through a FAK/SRC axis. During melanoma dissemination, NECTIN1 loss thus causes a cellular phenotypic switch from cell-cell adhesion to cell-matrix adhesion triggered by IGF1 signaling. Our study uncovers a mechanism by which cancer cells integrate information from the tumor microenvironment and cell-cell contacts to regulate their migratory behavior during spreading.
2022-05-26 | GSE149481 | GEO
Project description:Decline in hematopoietic function in aged individuals is associated with expansion of phenotypic hematopoietic stem cells (HSCs) and a shift in their lineage potential toward production of myeloid cells. Both HSC-intrinsic changes, and extrinsic changes in the bone marrow (BM) microenvironment, have been identified in old mice and humans. However, to extend healthy and robust hematopoietic function from youth into older age, we need to understand and effectively target the processes that initiate functional hematopoietic decline. We recently identified decline in Insulin-Like Growth Factor 1 (IGF1) in the BM microenvironment as early as middle age to be an HSC-extrinsic initiating driver of HSC aging (Young et al., Cell Stem Cell 2021). As systemic IGF1 administration has significant undesirable side effects, we sought to comprehensively interrogate the cell population(s) in the BM microenvironment that are responsible for IGF1 decline, towards the goal of cell type-specific targeted therapy. We performed single cell RNA-seq to comprehensively profile hematopoietic and non-hematopoietic fractions of the BM in young (2-4mo; n = 5 biological replicates) and middle-aged (12-14mo; n = 10) mice. In young mice, we find Igf1 to be nearly entirely detected in the mesenchymal stromal cell populations Adipo-CAR and Osteo-CAR, and Igf1 is significantly reduced in expression in both populations in middle-aged mice. Using two independent mesenchymal stromal cell Cre mouse lines, Lepr-Cre and Prx1-CreERT2, we found that knockout of Igf1 resulted in myeloid-biased hematopoiesis that replicated aging phenotypes. This result was similar to our published work showing that knockout of Igf1 using Nestin-CreER causes myeloid-biased hematopoiesis. While these Cre models generally do not mark similar cell types, it has been shown that Lepr-Cre-expressing perisinusoidal stromal cells include cells that express certain Nestin transgenes. Using fluorescent reporters, we find that all three lines (Lepr-Cre, Prx1-CreERT2, and Nestin-CreER) overlap in expression in the CAR populations that abundantly express Igf1 in young mice. Taken together, our work identifies a new role for Cxcl12-abundant reticular cells in sustaining hematopoietic function through local IGF1 production and suggests that specifically targeting CAR cells to maintain or restore Igf1 expression during aging will have beneficial effects on lymphoid cell production and adaptive immunity.
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