22 results on '"Torres-Collado AX"'
Search Results
2. Overcoming Resistance of Human Non-Hodgkin's Lymphoma to CD19-CAR CTL Therapy by Celecoxib and Histone Deacetylase Inhibitors.
- Author
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Torres-Collado AX and Jazirehi AR
- Abstract
Patients with B-cell non-Hodgkin’s lymphoma (B-NHL) who fail to respond to first-line treatment regimens or develop resistance, exhibit poor prognosis. This signifies the need to develop alternative treatment strategies. CD19-chimeric antigen receptor (CAR) T cell-redirected immunotherapy is an attractive and novel option, which has shown encouraging outcomes in phase I clinical trials of relapsed/refractory NHL. However, the underlying mechanisms of, and approaches to overcome, acquired anti-CD19CAR CD8⁺ T cells (CTL)-resistance in NHL remain elusive. CD19CAR transduced primary human CTLs kill CD19⁺ human NHLs in a CD19- and caspase-dependent manner, mainly via the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) apoptotic pathway. To understand the dynamics of the development of resistance, we analyzed several anti-CD19CAR CTL-resistant NHL sublines (R-NHL) derived by serial exposure of sensitive parental lines to excessive numbers of anti-CD19CAR CTLs followed by a limiting dilution analysis. The R-NHLs retained surface CD19 expression and were efficiently recognized by CD19CAR CTLs. However, R-NHLs developed cross-resistance to CD19CAR transduced human primary CTLs and the Jurkat human T cell line, activated Jurkat, and lymphokine activated killer (LAK) cells, suggesting the acquisition of resistance is independent of CD19-loss and might be due to aberrant apoptotic machinery. We hypothesize that the R-NHL refractoriness to CD19CAR CTL killing could be partially rescued by small molecule sensitizers with apoptotic-gene regulatory effects. Chromatin modifiers and Celecoxib partially reversed the resistance of R-NHL cells to the cytotoxic effects of anti-CD19CAR CTLs and rhTRAIL. These in vitro results, though they require further examination, may provide a rational biological basis for combination treatment in the management of CD19CAR CTL-based therapy of NHL.
- Published
- 2018
- Full Text
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3. Reversal of Resistance in Targeted Therapy of Metastatic Melanoma: Lessons Learned from Vemurafenib (BRAF V600E -Specific Inhibitor).
- Author
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Torres-Collado AX, Knott J, and Jazirehi AR
- Abstract
Malignant melanoma is the most aggressive form of skin cancer and has a very low survival rate. Over 50% of melanomas harbor various BRAF mutations with the most common being the V600E. BRAF
V600E mutation that causes constitutive activation of the MAPK pathway leading to drug-, immune-resistance, apoptosis evasion, proliferation, survival, and metastasis of melanomas. The ATP competitive BRAFV600E selective inhibitor, vemurafenib, has shown dramatic success in clinical trials; promoting tumor regression and an increase in overall survival of patients with metastatic melanoma. Regrettably, vemurafenib-resistance develops over an average of six months, which renders melanomas resistant to other therapeutic strategies. Elucidation of the underlying mechanism(s) of acquisition of vemurafenib-resistance and design of novel approaches to override resistance is the subject of intense clinical and basic research. In this review, we summarize recent developments in therapeutic approaches and clinical investigations on melanomas with BRAFV600E mutation to establish a new platform for the treatment of melanoma.- Published
- 2018
- Full Text
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4. Autocrine VEGF maintains endothelial survival through regulation of metabolism and autophagy.
- Author
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Domigan CK, Warren CM, Antanesian V, Happel K, Ziyad S, Lee S, Krall A, Duan L, Torres-Collado AX, Castellani LW, Elashoff D, Christofk HR, van der Bliek AM, Potente M, and Iruela-Arispe ML
- Subjects
- Animals, Blotting, Western, Cell Differentiation, Cell Proliferation, Cells, Cultured, Endothelium, Vascular metabolism, Forkhead Box Protein O1, Forkhead Transcription Factors genetics, Gene Expression Profiling, Humans, Hypoxia physiopathology, Mice, Mice, Knockout, Mitochondria metabolism, Phosphorylation, RNA, Messenger genetics, Real-Time Polymerase Chain Reaction, Reverse Transcriptase Polymerase Chain Reaction, Signal Transduction, Apoptosis, Autocrine Communication, Autophagy, Biomarkers metabolism, Endothelium, Vascular pathology, Forkhead Transcription Factors metabolism, Mitochondria pathology, Vascular Endothelial Growth Factor A physiology
- Abstract
Autocrine VEGF is necessary for endothelial survival, although the cellular mechanisms supporting this function are unknown. Here, we show that--even after full differentiation and maturation--continuous expression of VEGF by endothelial cells is needed to sustain vascular integrity and cellular viability. Depletion of VEGF from the endothelium results in mitochondria fragmentation and suppression of glucose metabolism, leading to increased autophagy that contributes to cell death. Gene-expression profiling showed that endothelial VEGF contributes to the regulation of cell cycle and mitochondrial gene clusters, as well as several--but not all--targets of the transcription factor FOXO1. Indeed, VEGF-deficient endothelium in vitro and in vivo showed increased levels of FOXO1 protein in the nucleus and cytoplasm. Silencing of FOXO1 in VEGF-depleted cells reversed expression profiles of several of the gene clusters that were de-regulated in VEGF knockdown, and rescued both cell death and autophagy phenotypes. Our data suggest that endothelial VEGF maintains vascular homeostasis through regulation of FOXO1 levels, thereby ensuring physiological metabolism and endothelial cell survival., (© 2015. Published by The Company of Biologists Ltd.)
- Published
- 2015
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5. Aberrant apoptotic machinery confers melanoma dual resistance to BRAF(V600E) inhibitor and immune effector cells: immunosensitization by a histone deacetylase inhibitor.
- Author
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Jazirehi AR, Nazarian R, Torres-Collado AX, and Economou JS
- Abstract
BRAF(V600E)-inhibitors (BRAFi; e.g., vemurafenib) and modern immune-based therapies such as PD-1/PD-L1 and CTLA-4 checkpoints blockade and adoptive cell transfer (ACT) have significantly improved the care of melanoma patients. Having these two effective (BRAFi and immunotherapy) therapies raises the question whether there is a rational biological basis for using them in combination. We developed an in vitro model to determine whether tumor resistance mechanisms to a small molecule inhibitor of a driver oncogene, and to cytotoxic T lymphocyte (CTL)- and natural killer (NK) cell-delivered apoptotic death signals were exclusive or intersecting. We generated melanoma sublines resistant to BRAFi vemurafenib and to CTL recognizing the MART-1 melanoma antigen. Vemurafenib-resistant (VemR) sublines were cross-resistant to MART CTL and NK cells indicating that a common apoptotic pathway governing tumor response to both modalities was disrupted. Pretreatment of VemR melanomas with a histone deacetylase inhibitor (HDACi) restored sensitivity to MART CTL and NK apoptosis by skewing the apoptotic gene programs towards a proapoptotic phenotype. Our in vitro findings suggest that during the course of acquisition of BRAFi resistance, melanomas develop cross-resistance to CTL- and NK-killing. Further, aberrant apoptotic pathways, amenable by an FDA-approved chromatin remodeling drug, regulate tumor resistance mechanisms to immune effector cells. These results may provide rational molecular basis for further investigations to combine these therapies clinically.
- Published
- 2014
6. Contribution of ADAMTS1 as a tumor suppressor gene in human breast carcinoma. Linking its tumor inhibitory properties to its proteolytic activity on nidogen-1 and nidogen-2.
- Author
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Martino-Echarri E, Fernández-Rodríguez R, Rodríguez-Baena FJ, Barrientos-Durán A, Torres-Collado AX, Plaza-Calonge Mdel C, Amador-Cubero S, Cortés J, Reynolds LE, Hodivala-Dilke KM, and Rodríguez-Manzaneque JC
- Subjects
- ADAM Proteins genetics, ADAMTS1 Protein, Animals, Basement Membrane metabolism, Basement Membrane pathology, Breast Neoplasms blood supply, Breast Neoplasms metabolism, Breast Neoplasms pathology, Calcium-Binding Proteins, Cell Adhesion Molecules genetics, Cell Line, Down-Regulation, Glycosaminoglycans genetics, Glycosaminoglycans metabolism, Glycosaminoglycans physiology, HEK293 Cells, Humans, Membrane Glycoproteins genetics, Mice, Mice, Inbred BALB C, Neovascularization, Pathologic genetics, Neovascularization, Pathologic metabolism, Neovascularization, Pathologic pathology, Proteolysis, Tumor Suppressor Proteins genetics, Tumor Suppressor Proteins metabolism, ADAM Proteins metabolism, Breast Neoplasms genetics, Cell Adhesion Molecules metabolism, Genes, Tumor Suppressor, Membrane Glycoproteins metabolism, Peptide Hydrolases metabolism
- Abstract
The extracellular protease ADAMTS1 (A disintegrin and metalloprotease with thrombospondin repeats 1) has been described as an anti-angiogenic molecule and its role as a putative tumor protective molecule has also been suggested. Here, we have used a tumor xenograft model to determine the role of ADAMTS1 in tumor growth and angiogenesis. Increasing levels of the protease led to the complete inhibition of tumor growth. In an attempt to elucidate the mechanism of action of this protease, we focused our attention on its proteolytic activity on nidogens, one of the main components of the vascular basement membrane. The increased expression of ADAMTS1 was accompanied by increased proteolysis of nidogen-1 and -2 and their almost complete removal from vascular structures, together with major morphological alterations of tumor blood vessels and a decreased vessel density. The clinical relevance of this work is supported by our observations that ADAMTS1 expression is decreased in breast tumor specimens when compared with healthy tissue. Our studies also reveal that the cleavage of nidogen-1 and -2 is partially inhibited in human tumor samples. Moreover, the deposition of both nidogens surrounding vascular structures is drastically altered, implying a possible reduction in the maintenance of vessel integrity. Our studies reflect the requirement to explore the functional interactions between proteases and specific substrates in cancer biology., (Copyright © 2013 UICC.)
- Published
- 2013
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7. Regulation of the apoptotic activity of the ING member of histone acetyltransferase and deacetylase complexes.
- Author
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Jazirehi AR and Torres-Collado AX
- Subjects
- Animals, Female, Humans, NF-kappa B metabolism, Pancreatic Neoplasms genetics, Pancreatic Neoplasms metabolism, SOX9 Transcription Factor genetics, SOX9 Transcription Factor metabolism
- Published
- 2013
8. Prevalence of DNA methyltransferase in breast cancer by SNP analysis.
- Author
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Jazirehi AR and Torres-Collado AX
- Published
- 2013
9. Epigenetic biomarker of metastatic melanoma.
- Author
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Torres-Collado AX and Jazirehi AR
- Subjects
- Humans, Biomarkers, Tumor blood, DNA, Neoplasm blood, Glycoproteins genetics, Melanoma diagnosis, Neoplasm Metastasis diagnosis, Skin Neoplasms diagnosis
- Published
- 2013
10. Epigenetic fine-tuning of NF-κB-Sox9 circuitry in cancer stem cells.
- Author
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Jazirehi AR and Torres-Collado AX
- Subjects
- Animals, Female, Humans, NF-kappa B metabolism, Pancreatic Neoplasms genetics, Pancreatic Neoplasms metabolism, SOX9 Transcription Factor genetics, SOX9 Transcription Factor metabolism
- Published
- 2013
11. Epigenetic regulation of retinoic acid susceptibility in leukemia.
- Author
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Torres-Collado AX and Jazirehi AR
- Published
- 2013
12. Role of miR-18b/MDM2/p53 circuitry in melanoma progression.
- Author
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Jazirehi AR, Torres-Collado AX, and Nazarian R
- Subjects
- Humans, Melanoma metabolism, MicroRNAs metabolism, Proto-Oncogene Proteins c-mdm2 metabolism, Signal Transduction, Skin Neoplasms metabolism, Tumor Suppressor Protein p53 metabolism
- Published
- 2013
13. Circumvention of drug-resistance via decitabine-mediated DNMT1 depletion.
- Author
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Torres-Collado AX and Jazirehi AR
- Subjects
- Female, Humans, Antimetabolites, Antineoplastic pharmacology, Azacitidine analogs & derivatives, Breast Neoplasms drug therapy, DNA (Cytosine-5-)-Methyltransferases antagonists & inhibitors, Drug Resistance, Neoplasm drug effects, Gene Expression Regulation, Neoplastic drug effects, Polyethylene Glycols, Polyethyleneimine
- Published
- 2013
14. Epigenetic regulation of melanoma tumor suppressor miRNA-124a.
- Author
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Jazirehi AR, Torres-Collado AX, and Nazarian R
- Subjects
- Humans, Epigenesis, Genetic physiology, Eye Neoplasms metabolism, Melanoma metabolism, MicroRNAs physiology, Tumor Suppressor Proteins physiology, Uveal Neoplasms metabolism
- Published
- 2013
15. Selective decline of synaptic protein levels in the frontal cortex of female mice deficient in the extracellular metalloproteinase ADAMTS1.
- Author
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Howell MD, Torres-Collado AX, Iruela-Arispe ML, and Gottschall PE
- Subjects
- ADAM Proteins metabolism, ADAMTS1 Protein, Animals, Cerebral Cortex metabolism, Female, Learning, Male, Memory, Mice, Neuronal Plasticity genetics, RNA, Messenger metabolism, Receptors, AMPA metabolism, ADAM Proteins genetics, Extracellular Matrix enzymology, Synapses metabolism
- Abstract
The chondroitin sulfate-bearing proteoglycans, also known as lecticans, are a major component of the extracellular matrix (ECM) in the central nervous system and regulate neural plasticity. Growing evidence indicates that endogenous, extracellular metalloproteinases that cleave lecticans mediate neural plasticity by altering the structure of ECM aggregates. The bulk of this in vivo data examined the matrix metalloproteinases, but another metalloproteinase family that cleaves lecticans, a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS), modulates structural plasticity in vitro, although few in vivo studies have tested this concept. Thus, the purpose of this study was to examine the neurological phenotype of a mouse deficient in ADAMTS1. Adamts1 mRNA was absent in the ADAMTS1 null mouse frontal cortex, but there was no change in the abundance or proteolytic processing of the prominent lecticans brevican and versican V2. However, there was a marked increase in the perinatal lectican neurocan in juvenile ADAMTS1 null female frontal cortex. More prominently, there were declines in synaptic protein levels in the ADAMTS1 null female, but not male, frontal cortex beginning at postnatal day 28. These synaptic marker declines did not affect learning or memory in the adult female ADAMTS1 null mice when tested with the radial-arm water maze. These results indicate that in vivo Adamts1 knockout leads to sexual dimorphism in frontal cortex synaptic protein levels. Since changes in lectican abundance and proteolytic processing did not accompany the synaptic protein declines, ADAMTS1 may play a nonproteolytic role in regulating neural plasticity.
- Published
- 2012
- Full Text
- View/download PDF
16. ADAMTS1 contributes to the acquisition of an endothelial-like phenotype in plastic tumor cells.
- Author
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Casal C, Torres-Collado AX, Plaza-Calonge Mdel C, Martino-Echarri E, Ramón Y Cajal S, Rojo F, Griffioen AW, and Rodríguez-Manzaneque JC
- Subjects
- ADAM Proteins antagonists & inhibitors, ADAM Proteins metabolism, ADAMTS1 Protein, Animals, Cell Line, Tumor, Endothelial Cells enzymology, Endothelial Cells pathology, Fibrosarcoma enzymology, Fibrosarcoma pathology, Humans, Mice, Mice, Inbred BALB C, Phenotype, Transplantation, Heterologous, ADAM Proteins biosynthesis, Melanoma enzymology, Melanoma pathology, Sarcoma, Ewing enzymology, Sarcoma, Ewing pathology
- Abstract
Cancer stem cells have been hypothesized to explain tumor plasticity, including the capability to adopt distinct differentiation commitments. Among the mechanisms of tumor neovascularization, the ability of some malignant cells to mimic an endothelial phenotype has been recognized by a capacity to form matrix-enriched pseudovascular structures. In addition to the expression of genes associated with an endothelial nature, the molecular dynamism of specific microenvironments may also be critical. Here, we report the identification of the extracellular protease ADAMTS1 as a critical molecule for tumor cells to acquire endothelial-like properties. In a fibrosarcoma model, ADAMTS1 increased tumor growth rate in an angiogenesis-independent manner, influencing the tumor cells to display an exclusive endothelial-like gene signature. We documented the relevant expression of ADAMTS1 in aggressive and highly plastic melanoma and Ewing sarcoma cells. Notably, inhibiting ADAMTS1 action compromised the endothelial mimetic attributes observed in this setting. Our findings provide insights into how the tumor microenvironment can elicit endothelial mimicry by tumor cells., (Copyright 2010 AACR.)
- Published
- 2010
- Full Text
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17. Loss of myeloid cell-derived vascular endothelial growth factor accelerates fibrosis.
- Author
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Stockmann C, Kerdiles Y, Nomaksteinsky M, Weidemann A, Takeda N, Doedens A, Torres-Collado AX, Iruela-Arispe L, Nizet V, and Johnson RS
- Subjects
- Animals, Bleomycin toxicity, Female, Hypoxia pathology, Mice, Mice, Mutant Strains, Phosphorylation, Pulmonary Fibrosis chemically induced, Pulmonary Fibrosis pathology, Vascular Endothelial Growth Factor A genetics, Vascular Endothelial Growth Factor A metabolism, Vascular Endothelial Growth Factor Receptor-2 metabolism, beta Catenin metabolism, Gene Deletion, Pulmonary Fibrosis genetics, Vascular Endothelial Growth Factor A physiology
- Abstract
Tissue injury initiates a complex series of events that act to restore structure and physiological homeostasis. Infiltration of inflammatory cells and vascular remodeling are both keystones of this process. However, the role of inflammation and angiogenesis in general and, more specifically, the significance of inflammatory cell-derived VEGF in this context are unclear. To determine the role of inflammatory cell-derived VEGF in a clinically relevant and chronically inflamed injury, pulmonary fibrosis, we deleted the VEGF-A gene in myeloid cells. In a model of pulmonary fibrosis in mice, deletion of VEGF in myeloid cells resulted in significantly reduced formation of blood vessels; however, it causes aggravated fibrotic tissue damage. This was accompanied by a pronounced decrease in epithelial cell survival and a striking increase in myofibroblast invasion. The drastic increase in fibrosis following loss of myeloid VEGF in the damaged lungs was also marked by increased levels of hypoxia-inducible factor (HIF) expression and Wnt/beta-catenin signaling. This demonstrates that the process of angiogenesis, driven by myeloid cell-derived VEGF, is essential for the prevention of fibrotic damage.
- Published
- 2010
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18. Targeting distinct tumor-infiltrating myeloid cells by inhibiting CSF-1 receptor: combating tumor evasion of antiangiogenic therapy.
- Author
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Priceman SJ, Sung JL, Shaposhnik Z, Burton JB, Torres-Collado AX, Moughon DL, Johnson M, Lusis AJ, Cohen DA, Iruela-Arispe ML, and Wu L
- Subjects
- Animals, Carcinoma, Lewis Lung pathology, Cell Line, Tumor, Lung Neoplasms pathology, Macrophages cytology, Male, Matrix Metalloproteinase 9 metabolism, Melanoma drug therapy, Melanoma pathology, Mice, Mice, Inbred C57BL, Myeloid Cells drug effects, Myeloid Cells pathology, Neoplasm Transplantation, Neovascularization, Pathologic metabolism, Prostatic Neoplasms drug therapy, Prostatic Neoplasms pathology, Rats, Receptor, Macrophage Colony-Stimulating Factor metabolism, Signal Transduction drug effects, Skin Neoplasms drug therapy, Skin Neoplasms pathology, Vascular Endothelial Growth Factor Receptor-2 metabolism, Anisoles pharmacology, Carcinoma, Lewis Lung drug therapy, Cell Movement drug effects, Lung Neoplasms drug therapy, Neovascularization, Pathologic drug therapy, Pyrimidines pharmacology, Receptor, Macrophage Colony-Stimulating Factor antagonists & inhibitors
- Abstract
Tumor-infiltrating myeloid cells (TIMs) support tumor growth by promoting angiogenesis and suppressing antitumor immune responses. CSF-1 receptor (CSF1R) signaling is important for the recruitment of CD11b(+)F4/80(+) tumor-associated macrophages (TAMs) and contributes to myeloid cell-mediated angiogenesis. However, the impact of the CSF1R signaling pathway on other TIM subsets, including CD11b(+)Gr-1(+) myeloid-derived suppressor cells (MDSCs), is unknown. Tumor-infiltrating MDSCs have also been shown to contribute to tumor angiogenesis and have recently been implicated in tumor resistance to antiangiogenic therapy, yet their precise involvement in these processes is not well understood. Here, we use the selective pharmacologic inhibitor of CSF1R signaling, GW2580, to demonstrate that CSF-1 regulates the tumor recruitment of CD11b(+)Gr-1(lo)Ly6C(hi) mononuclear MDSCs. Targeting these TIM subsets inhibits tumor angiogenesis associated with reduced expression of proangiogenic and immunosuppressive genes. Combination therapy using GW2580 with an anti-VEGFR-2 antibody synergistically suppresses tumor growth and severely impairs tumor angiogenesis along with reverting at least one TIM-mediated antiangiogenic compensatory mechanism involving MMP-9. These data highlight the importance of CSF1R signaling in the recruitment and function of distinct TIM subsets, including MDSCs, and validate the benefits of targeting CSF1R signaling in combination with antiangiogenic drugs for the treatment of solid cancers.
- Published
- 2010
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19. RETRACTED: Variable inhibition of thrombospondin 1 against liver and lung metastases through differential activation of metalloproteinase ADAMTS1.
- Author
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Lee YJ, Koch M, Karl D, Torres-Collado AX, Fernando NT, Rothrock C, Kuruppu D, Ryeom S, Iruela-Arispe ML, and Yoon SS
- Subjects
- ADAM Proteins genetics, ADAMTS1 Protein, Animals, Binding Sites, Blotting, Western, Cell Line, Cell Line, Tumor, Cell Proliferation, Endothelial Cells metabolism, Endothelial Cells pathology, Female, Humans, Liver Neoplasms genetics, Liver Neoplasms metabolism, Lung Neoplasms genetics, Lung Neoplasms metabolism, Male, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, Mice, Knockout, Neoplasms, Experimental genetics, Neoplasms, Experimental metabolism, RNA Interference, Reverse Transcriptase Polymerase Chain Reaction, Thrombospondin 1 genetics, Transfection, ADAM Proteins metabolism, Liver Neoplasms secondary, Lung Neoplasms secondary, Neoplasms, Experimental pathology, Thrombospondin 1 metabolism
- Abstract
Metastasis relies on angiogenesis for tumor expansion. Tumor angiogenesis is restrained by a variety of endogenous inhibitors, including thrombospondin 1 (TSP1). The principal antiangiogenic activity of TSP1 resides in a domain containing three TSP1 repeats (3TSR), and TSP1 cleavage is regulated, in part, by the metalloproteinase ADAMTS1. In this study, we examined the role of TSP1 and ADAMTS1 in controlling metastatic disease in the liver and lung. TSP1 overexpression inhibited metastatic growth of colon or renal carcinoma cells in liver but not lung. Metastatic melanoma in liver grew more rapidly in Tsp1-null mice compared with controls, whereas in lung grew similarly in Tsp1-null mice or controls. Recombinant TSP1 was cleaved more efficiently in lysates from liver than lung. ADAMTS1 inhibition by neutralizing antibody, small interfering RNA, or genetic deletion abrogated cleavage activity. To confirm that lack of cleavage of TSP1 ablated its antiangiogenic function in the lung, we generated colon cancer cells stably secreting only the 3TSR domain and found that they inhibited formation of both liver and lung metastases. Collectively, our results indicate that the antiangiogenic activity of TSP1 is differentially regulated by ADAMTS1 in the liver and lung, emphasizing the concept that regulation of angiogenesis is varied in different tissue environments.
- Published
- 2010
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20. Cleavage of syndecan-4 by ADAMTS1 provokes defects in adhesion.
- Author
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Rodríguez-Manzaneque JC, Carpizo D, Plaza-Calonge Mdel C, Torres-Collado AX, Thai SN, Simons M, Horowitz A, and Iruela-Arispe ML
- Subjects
- ADAMTS1 Protein, ADAMTS4 Protein, Actins metabolism, Animals, Binding Sites, CHO Cells, Cell Adhesion physiology, Cell Line, Cell Movement physiology, Cricetinae, Cricetulus, Focal Adhesions metabolism, Glycosaminoglycans metabolism, Metalloproteases genetics, Metalloproteases metabolism, Mice, Procollagen N-Endopeptidase metabolism, Transfection, ADAM Proteins metabolism, Membrane Glycoproteins metabolism, Proteoglycans metabolism, Syndecan-4 metabolism
- Abstract
Syndecan-4 is a membrane-bound heparan sulfate proteoglycan that participates in cell-cell and cell-matrix interactions and modulates adhesion and migration of many cell types. Through its extracellular domain, syndecan-4 cooperates with adhesion molecules and binds matrix components relevant for cell migration. Importantly, syndecan-4 is a substrate of extracellular proteases, however the biological significance of this cleavage has not been elucidated. Here, we show that the secreted metalloprotease ADAMTS1, involved in angiogenesis and inflammatory processes, cleaves the ectodomain of syndecan-4. We further showed that this cleavage results in altered distribution of cytoskeleton components, functional loss of adhesion, and gain of migratory capacities. Using syndecan-4 null cells, we observed that ADAMTS1 proteolytic action mimics the outcome of genetic deletion of this proteoglycan with regards to focal adhesion. Our findings suggest that the shedding of syndecan-4 by ADAMTS1 disrupts cell adhesion and promotes cell migration.
- Published
- 2009
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21. ADAMTS1 interacts with, cleaves, and modifies the extracellular location of the matrix inhibitor tissue factor pathway inhibitor-2.
- Author
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Torres-Collado AX, Kisiel W, Iruela-Arispe ML, and Rodríguez-Manzaneque JC
- Subjects
- ADAM Proteins genetics, ADAMTS1 Protein, Animals, Cell Line, Tumor, Extracellular Matrix Proteins chemistry, Extracellular Matrix Proteins genetics, Gene Library, Glycoproteins chemistry, Glycoproteins genetics, Humans, Kidney Neoplasms, Mice, Mice, Inbred BALB C, Mice, Nude, Neoplasm Transplantation, Protein Structure, Tertiary, Respiratory Mucosa cytology, Respiratory Mucosa metabolism, Transplantation, Heterologous, Two-Hybrid System Techniques, ADAM Proteins metabolism, Extracellular Matrix Proteins metabolism, Glycoproteins metabolism
- Abstract
ADAMTS1 is an extracellular metalloproteinase known to participate in a variety of biological processes that includes inflammation, angiogenesis, and development of the urogenital system. Many of its functions rely on its catalytic activity, which thus far has been limited to the cleavage of the matrix proteoglycans aggrecan and versican. However, it is likely that other substrates exist. Using a yeast two-hybrid screen, we identified the Kunitz-type inhibitor, tissue factor pathway inhibitor-2 (TFPI-2), as a binding partner of ADAMTS1. The interaction was confirmed by several biochemical and cell-based assays. In addition, our studies revealed alterations in the pattern of TFPI-2-secreted isoforms and in its extracellular location caused by the specific action of ADAMTS1. Interestingly, we found that TFPI-2 is a novel substrate of ADAMTS1. The cleavage removes a protease-sensitive C-terminal region in TFPI-2, altering its binding properties. The proposed role of TFPI-2 as a maintenance factor of extracellular remodeling suggests the indirect function of ADAMTS1 as an additional homeostatic player by its ability to alter the extracellular location of TFPI-2 and, therefore, to disrupt the remodeling machinery, a phenomenon directly associated to pathologies such as atherosclerosis and tumor progression.
- Published
- 2006
- Full Text
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22. Anti-tRNP(ser)sec/SLA/LP autoantibodies. Comparative study using in-house ELISA with a recombinant 48.8 kDa protein, immunoblot, and analysis of immunoprecipitated RNAs.
- Author
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Torres-Collado AX, Czaja AJ, and Gelpí C
- Subjects
- Enzyme-Linked Immunosorbent Assay, Female, Hepatitis, Autoimmune physiopathology, Hepatitis, Chronic physiopathology, Humans, Immunoblotting, Male, Probability, Recombinant Proteins genetics, Ribonucleoproteins analysis, Ribonucleoproteins immunology, Sampling Studies, Sensitivity and Specificity, Autoantibodies analysis, Hepatitis, Autoimmune immunology, Hepatitis, Chronic immunology, Recombinant Proteins immunology
- Abstract
Background: Antibodies against tRNP((ser)sec) (ribonucleoproteins, RNP) have been described in our laboratory as markers of poor outcome in type 1 autoimmune hepatitis (AIH). The antigenic protein has been sequenced and cloned as a 48.8 kDa protein and identified with soluble liver antigen (SLA) and liver-pancreas (LP) antigen. The aim of this paper was to determine the best assay by which to detect these antibodies in type 1 AIH., Methods: A simple and reliable enzyme linked immunoassay based on prokaryotically expressed protein was compared with an immunoblot assay using prokaryotically- and eukaryotically-expressed proteins and an assay based on immunoprecipitated RNAs from HeLa cell extracts. Eighty-one sera from 58 patients with type 1 AIH, 168 sera from patients with autoimmune diseases or chronic hepatitis C, and 60 sera from healthy subjects were similarly tested., Results: The specificity of the assays was 100%, but the frequency of seropositivity was higher in the assay based on immunoprecipitated RNAs (44.4%) than in the enzyme-linked immunosorbent assay (ELISA) (16%) and the immunoblot assay with prokaryotically (12.34%) and eukaryotically (14.8%)-expressed protein. There were no clinical differences between the patients positive by ELISA, immunoblot assay, or immunoprecipitated RNAs., Conclusions: These results suggest that the analysis of the immunoprecipitated RNAs is the most useful, sensitive and specific method to detect anti-tRNP((ser)sec)/SLA/LP autoantibodies., (Copyright Blackwell Munksgaard 2005)
- Published
- 2005
- Full Text
- View/download PDF
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