|
1
|
Penkov D, Beloglazova I, Parfyonova Y. Endothelial-specific Enhancer as a Cis Element of PLAUR Regulation by TNF-alpha, IL-1beta, and VEGF. Curr Pharm Des 2024; 30:1630-1640. [PMID: 38715331 DOI: 10.2174/0113816128296376240424072322] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/06/2024] [Accepted: 04/03/2024] [Indexed: 01/23/2025]
Abstract
The expression of human PLAUR gene, which encodes the urokinase plasminogen activator receptor (uPAR), is cell- and process-specific and elevated in inflammation, cancer and senescence. Its tight regulation is achieved by regulatory elements in the gene locus, such as the promoter and several enhancers. The promoter activity is not specific to a particular cell type and has been described earlier. The proximal enhancer is endothelial-specific and responsible for the PLAUR expression pattern in endothelial cells. In this study we described the enhancer activity and its cis-regulatory elements based on the published data. We showed a possible connection of the enhancer activity with known cellular phenotypes.
Collapse
Affiliation(s)
- Dmitry Penkov
- Laboratory of Angiogenesis, Institute of Experimental Cardiology Named after Academician V.N. Smirnov, Federal State Budgetary Institution National Medical Research Center of Cardiology Named after Academician E.I. Chazov, Ministry of Health of the Russian Federation, Moscow, Russia
| | - Irina Beloglazova
- Laboratory of Angiogenesis, Institute of Experimental Cardiology Named after Academician V.N. Smirnov, Federal State Budgetary Institution National Medical Research Center of Cardiology Named after Academician E.I. Chazov, Ministry of Health of the Russian Federation, Moscow, Russia
| | - Yelena Parfyonova
- Laboratory of Angiogenesis, Institute of Experimental Cardiology Named after Academician V.N. Smirnov, Federal State Budgetary Institution National Medical Research Center of Cardiology Named after Academician E.I. Chazov, Ministry of Health of the Russian Federation, Moscow, Russia
| |
Collapse
|
|
2
|
Therapeutic Strategies Targeting Urokinase and Its Receptor in Cancer. Cancers (Basel) 2022; 14:cancers14030498. [PMID: 35158766 PMCID: PMC8833673 DOI: 10.3390/cancers14030498] [Citation(s) in RCA: 30] [Impact Index Per Article: 10.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/18/2021] [Revised: 01/11/2022] [Accepted: 01/15/2022] [Indexed: 01/19/2023] Open
Abstract
Several studies have ascertained that uPA and uPAR do participate in tumor progression and metastasis and are involved in cell adhesion, migration, invasion and survival, as well as angiogenesis. Increased levels of uPA and uPAR in tumor tissues, stroma and biological fluids correlate with adverse clinic-pathologic features and poor patient outcomes. After binding to uPAR, uPA activates plasminogen to plasmin, a broad-spectrum matrix- and fibrin-degrading enzyme able to facilitate tumor cell invasion and dissemination to distant sites. Moreover, uPAR activated by uPA regulates most cancer cell activities by interacting with a broad range of cell membrane receptors. These findings make uPA and uPAR not only promising diagnostic and prognostic markers but also attractive targets for developing anticancer therapies. In this review, we debate the uPA/uPAR structure-function relationship as well as give an update on the molecules that interfere with or inhibit uPA/uPAR functions. Additionally, the possible clinical development of these compounds is discussed.
Collapse
|
|
3
|
Recent Applications of Retro-Inverso Peptides. Int J Mol Sci 2021; 22:ijms22168677. [PMID: 34445382 PMCID: PMC8395423 DOI: 10.3390/ijms22168677] [Citation(s) in RCA: 43] [Impact Index Per Article: 10.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/28/2021] [Revised: 08/09/2021] [Accepted: 08/10/2021] [Indexed: 12/14/2022] Open
Abstract
Natural and de novo designed peptides are gaining an ever-growing interest as drugs against several diseases. Their use is however limited by the intrinsic low bioavailability and poor stability. To overcome these issues retro-inverso analogues have been investigated for decades as more stable surrogates of peptides composed of natural amino acids. Retro-inverso peptides possess reversed sequences and chirality compared to the parent molecules maintaining at the same time an identical array of side chains and in some cases similar structure. The inverted chirality renders them less prone to degradation by endogenous proteases conferring enhanced half-lives and an increased potential as new drugs. However, given their general incapability to adopt the 3D structure of the parent peptides their application should be careful evaluated and investigated case by case. Here, we review the application of retro-inverso peptides in anticancer therapies, in immunology, in neurodegenerative diseases, and as antimicrobials, analyzing pros and cons of this interesting subclass of molecules.
Collapse
|
|
4
|
Stimuli responsive and receptor targeted iron oxide based nanoplatforms for multimodal therapy and imaging of cancer: Conjugation chemistry and alternative therapeutic strategies. J Control Release 2021; 333:188-245. [DOI: 10.1016/j.jconrel.2021.03.021] [Citation(s) in RCA: 16] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/05/2020] [Revised: 03/17/2021] [Accepted: 03/17/2021] [Indexed: 12/18/2022]
|
|
5
|
Napolitano F, Montuori N. The Role of the Plasminogen Activation System in Angioedema: Novel Insights on the Pathogenesis. J Clin Med 2021; 10:518. [PMID: 33535668 PMCID: PMC7867209 DOI: 10.3390/jcm10030518] [Citation(s) in RCA: 28] [Impact Index Per Article: 7.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/22/2020] [Revised: 01/22/2021] [Accepted: 01/28/2021] [Indexed: 12/21/2022] Open
Abstract
The main physiological functions of plasmin, the active form of its proenzyme plasminogen, are blood clot fibrinolysis and restoration of normal blood flow. The plasminogen activation (PA) system includes urokinase-type plasminogen activator (uPA), tissue-type PA (tPA), and two types of plasminogen activator inhibitors (PAI-1 and PAI-2). In addition to the regulation of fibrinolysis, the PA system plays an important role in other biological processes, which include degradation of extracellular matrix such as embryogenesis, cell migration, tissue remodeling, wound healing, angiogenesis, inflammation, and immune response. Recently, the link between PA system and angioedema has been a subject of scientific debate. Angioedema is defined as localized and self-limiting edema of subcutaneous and submucosal tissues, mediated by bradykinin and mast cell mediators. Different forms of angioedema are linked to uncontrolled activation of coagulation and fibrinolysis systems. Moreover, plasmin itself can induce a potentiation of bradykinin production with consequent swelling episodes. The number of studies investigating the PA system involvement in angioedema has grown in recent years, highlighting its relevance in etiopathogenesis. In this review, we present the components and diverse functions of the PA system in physiology and its importance in angioedema pathogenesis.
Collapse
Affiliation(s)
| | - Nunzia Montuori
- Department of Translational Medical Sciences and Center for Basic and Clinical Immunology Research (CISI), University of Naples Federico II, 80135 Naples, Italy;
| |
Collapse
|
|
6
|
Chu Y, Bucci JC, Peterson CB. Identification of a PAI-1-binding site within an intrinsically disordered region of vitronectin. Protein Sci 2019; 29:494-508. [PMID: 31682300 DOI: 10.1002/pro.3770] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/20/2019] [Accepted: 10/28/2019] [Indexed: 12/14/2022]
Abstract
The serine protease inhibitor, plasminogen activator inhibitor Type-1 (PAI-1) is a metastable protein that undergoes an unusual transition to an inactive conformation with a short half-life of only 1-2 hr. Circulating PAI-1 is bound to a cofactor vitronectin, which stabilizes PAI-1 by slowing this latency conversion. A well-characterized PAI-1-binding site on vitronectin is located within the somatomedin B (SMB) domain, corresponding to the first 44 residues of the protein. Another PAI-1 recognition site has been identified with an engineered form of vitronectin lacking the SMB domain, yet retaining PAI-1 binding capacity (Schar, Blouse, Minor, Peterson. J Biol Chem. 2008;283:28487-28496). This additional binding site is hypothesized to lie within an intrinsically disordered domain (IDD) of vitronectin. To localize the putative binding site, we constructed a truncated form of vitronectin containing 71 amino acids from the N-terminus, including the SMB domain and an additional 24 amino acids from the IDD region. This portion of the IDD is rich in acidic amino acids, which are hypothesized to be complementary to several basic residues identified within an extensive vitronectin-binding site mapped on PAI-1 (Schar, Jensen, Christensen, Blouse, Andreasen, Peterson. J Biol Chem. 2008;283:10297-10309). Steady-state and stopped-flow fluorescence measurements demonstrate that the truncated form of vitronectin exhibits the same rapid biphasic association as full-length vitronectin and that the IDD hosts the elusive second PAI-1 binding site that lies external to the SMB domain of vitronectin.
Collapse
Affiliation(s)
- Yuzhuo Chu
- Department of Biological Sciences, Louisiana State University, Baton Rouge, Louisiana, United States
| | - Joel C Bucci
- Department of Biological Sciences, Louisiana State University, Baton Rouge, Louisiana, United States
| | - Cynthia B Peterson
- Department of Biological Sciences, Louisiana State University, Baton Rouge, Louisiana, United States
| |
Collapse
|
|
7
|
Minopoli M, Polo A, Ragone C, Ingangi V, Ciliberto G, Pessi A, Sarno S, Budillon A, Costantini S, Carriero MV. Structure-function relationship of an Urokinase Receptor-derived peptide which inhibits the Formyl Peptide Receptor type 1 activity. Sci Rep 2019; 9:12169. [PMID: 31434916 PMCID: PMC6704176 DOI: 10.1038/s41598-019-47900-3] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/21/2019] [Accepted: 07/23/2019] [Indexed: 12/17/2022] Open
Abstract
The interaction between the short 88Ser-Arg-Ser-Arg-Tyr92 sequence of the urokinase receptor (uPAR) and the formyl peptide receptor type 1 (FPR1) elicits cell migration. We generated the Ac-(D)-Tyr-(D)-Arg-Aib-(D)-Arg-NH2 (RI-3) peptide which inhibits the uPAR/FPR1 interaction, reducing migration of FPR1 expressing cells toward N-formyl-methionyl-leucyl-phenylalanine (fMLF) and Ser-Arg-Ser-Arg-Tyr (SRSRY) peptides. To understand the structural basis of the RI-3 inhibitory effects, the FPR1/fMLF, FPR1/SRSRY and FPR1/RI-3 complexes were modeled and analyzed, focusing on the binding pocket of FPR1 and the interaction between the amino acids that signal to the FPR1 C-terminal loop. We found that RI-3 shares the same binding site of fMLF and SRSRY on FPR1. However, while fMLF and SRSRY display the same agonist activation signature (i.e. the series of contacts that transmit the conformational transition throughout the complex), translating binding into signaling, RI-3 does not interact with the activation region of FPR1 and hence does not activate signaling. Indeed, fluorescein-conjugated RI-3 prevents either fMLF and SRSRY uptake on FPR1 without triggering FPR1 internalization and cell motility in the absence of any stimulus. Collectively, our data show that RI-3 is a true FPR1 antagonist and suggest a pharmacophore model useful for development of compounds that selectively inhibit the uPAR-triggered, FPR1-mediated cell migration.
Collapse
Affiliation(s)
- Michele Minopoli
- Neoplastic Progression Unit, Istituto Nazionale Tumori IRCCS "Fondazione G. Pascale", Naples, Italy
| | - Andrea Polo
- Experimental Pharmacology Unit, Istituto Nazionale Tumori IRCCS "Fondazione G. Pascale", Napoli, Italy
| | - Concetta Ragone
- Neoplastic Progression Unit, Istituto Nazionale Tumori IRCCS "Fondazione G. Pascale", Naples, Italy
| | - Vincenzo Ingangi
- Immunologia e Diagnostica molecolare Istituto Oncologico Veneto, Padova, Italy
| | - Gennaro Ciliberto
- Scientific Directorate, Istituto Nazionale Tumori "Regina Elena", IRCCS, Roma, Italy
| | | | - Sabrina Sarno
- Neoplastic Progression Unit, Istituto Nazionale Tumori IRCCS "Fondazione G. Pascale", Naples, Italy
| | - Alfredo Budillon
- Experimental Pharmacology Unit, Istituto Nazionale Tumori IRCCS "Fondazione G. Pascale", Napoli, Italy
| | - Susan Costantini
- Experimental Pharmacology Unit, Istituto Nazionale Tumori IRCCS "Fondazione G. Pascale", Napoli, Italy.
| | - Maria Vincenza Carriero
- Neoplastic Progression Unit, Istituto Nazionale Tumori IRCCS "Fondazione G. Pascale", Naples, Italy.
| |
Collapse
|
|
8
|
Zhang ZT, Huang-Fu MY, Xu WH, Han M. Stimulus-responsive nanoscale delivery systems triggered by the enzymes in the tumor microenvironment. Eur J Pharm Biopharm 2019; 137:122-130. [PMID: 30776412 DOI: 10.1016/j.ejpb.2019.02.009] [Citation(s) in RCA: 46] [Impact Index Per Article: 7.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/05/2018] [Revised: 02/02/2019] [Accepted: 02/14/2019] [Indexed: 12/12/2022]
Abstract
The tumor microenvironment is the cellular environment that is also described as the "soil" for supporting tumor growth, proliferation, invasion and metastasis, as well as protecting tumor cells from immunological recognition. Notably, tumor cells can grow much faster than other normal organs and invade surrounding tissues more easily, which results in abnormal expression of enzymes in the tumor microenvironment, including matrix metalloproteinases, cathepsins, phospholipases, oxidoreductases, etc. In opposite, due to the high selectivity and catalytic activity, these enzymes can promote nanoparticles to recognize tumor tissues more accurately, and the more accumulation of drugs at primal tumor sites will enhance therapeutic efficacy with lower systemic toxicity. Therefore, one promising antitumor strategy is to design stimulus-responsive nanoscale delivery systems triggered by the enzymes with the support of various nanocarriers, such as liposomes, micelles and inorganic nanoparticles, etc. In this review, numerous facts were cited to summarize and discuss the typical types of enzyme-stimulus responsive nanoscale delivery systems. More importantly, we also focused on their recent advancements in antitumor therapy, and offered the direction for further studies.
Collapse
Affiliation(s)
- Zhen-Tao Zhang
- Institute of Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China
| | - Ming-Yi Huang-Fu
- Institute of Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China
| | - Wen-Hong Xu
- Department of Radiation Oncology, Key Laboratory of Cancer Prevention and Intervention, The Second Affiliated Hospital, Zhejiang University, College of Medicine, Hangzhou 310058 China.
| | - Min Han
- Institute of Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
| |
Collapse
|
|
9
|
Yousif AM, Ingangi V, Merlino F, Brancaccio D, Minopoli M, Bellavita R, Novellino E, Carriero MV, Carotenuto A, Grieco P. Urokinase receptor derived peptides as potent inhibitors of the formyl peptide receptor type 1-triggered cell migration. Eur J Med Chem 2017; 143:348-360. [PMID: 29202399 DOI: 10.1016/j.ejmech.2017.11.030] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/07/2017] [Revised: 11/09/2017] [Accepted: 11/11/2017] [Indexed: 10/18/2022]
Abstract
The receptor for the urokinase-type plasminogen activator (uPAR) is a widely recognized master regulator of cell migration. We and others have previously documented that the uPAR(84-95) sequence, interacts with the formyl peptide receptors (FPR)s, henceforth inducing cell migration of several cell lines, including leukocytes, and the synthetic shorter peptide (Ser88-Arg-Ser-Arg-Tyr92, SRSRY) retains chemotactic activity in vitro and in vivo. Recently, we have developed the head-to-tail cyclic analog [SRSRY], a new potent and stable inhibitor of monocyte trafficking. This prompted us to develop novel cyclic and linear analogs of [SRSRY] with the aim to broaden the knowledge about structure-activity relationships of peptide [SRSRY]. Herein we report their synthesis, effects on cell migration, conformational and docking analyses which served to envisage a new pharmacophore model for inhibitors of FPR1-triggered cell migration.
Collapse
Affiliation(s)
- Ali Munaim Yousif
- Department of Pharmacy, University of Naples 'Federico II', Naples 80131, Italy; Department of Chemistry, University of Texas at Dallas, 800 W. Campbell Rd., Richardson, TX 75080, United States
| | - Vincenzo Ingangi
- Department of Experimental Oncology IRCCS Istituto Nazionale Tumori "Fondazione G. Pascale" I-80131 Naples, Italy; Department of Experimental Medicine, University of Campania 'Luigi Vanvitelli', Naples 80138, Italy
| | - Francesco Merlino
- Department of Pharmacy, University of Naples 'Federico II', Naples 80131, Italy
| | - Diego Brancaccio
- Department of Pharmacy, University of Naples 'Federico II', Naples 80131, Italy
| | - Michele Minopoli
- Department of Experimental Oncology IRCCS Istituto Nazionale Tumori "Fondazione G. Pascale" I-80131 Naples, Italy; Department of Experimental Medicine, University of Campania 'Luigi Vanvitelli', Naples 80138, Italy
| | - Rosa Bellavita
- Department of Pharmacy, University of Naples 'Federico II', Naples 80131, Italy
| | - Ettore Novellino
- Department of Pharmacy, University of Naples 'Federico II', Naples 80131, Italy
| | - Maria Vincenza Carriero
- Department of Experimental Oncology IRCCS Istituto Nazionale Tumori "Fondazione G. Pascale" I-80131 Naples, Italy.
| | - Alfonso Carotenuto
- Department of Pharmacy, University of Naples 'Federico II', Naples 80131, Italy.
| | - Paolo Grieco
- Department of Pharmacy, University of Naples 'Federico II', Naples 80131, Italy; Centro Interuniversitario di Ricerca sui Peptidi Bioattivi (CIRPEB) University of Naples "Federico II" and DFM-Scarl, Institute of Biostructures and Bioimaging - CNR Via Mezzocannone 16, 80134 Naples, Italy
| |
Collapse
|
|
10
|
Retro-inverso Urokinase Receptor Antagonists for the Treatment of Metastatic Sarcomas. Sci Rep 2017; 7:1312. [PMID: 28465589 PMCID: PMC5430962 DOI: 10.1038/s41598-017-01425-9] [Citation(s) in RCA: 16] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/14/2016] [Accepted: 03/29/2017] [Indexed: 11/12/2022] Open
Abstract
The development of metastases is a multistep process that requires the activation of physiological and biochemical processes that govern migration, invasion and entry of metastatic cells into blood vessels. The urokinase receptor (uPAR) promotes cell migration by interacting with the Formyl Peptide Receptors (FPRs). Since both uPAR and FPR1 are involved in tumor progression, the uPAR-FPR1 interaction is an attractive therapeutic target. We previously described peptide antagonists of the uPAR-FPR1 interaction that inhibited cell migration and angiogenesis. To develop enzyme-resistant analogues, we applied here the Retro-Inverso (RI) approach, whereby the topology of the side chains is maintained by inverting the sequence of the peptide and the chirality of all residues. Molecular dynamics suggests that peptide RI-3 adopts the turn structure typical of uPAR-FPR1 antagonists. Accordingly, RI-3 is a nanomolar competitor of N-formyl-Met-Leu-Phe for binding to FPR1 and inhibits migration, invasion, trans-endothelial migration of sarcoma cells and VEGF-triggered endothelial tube formation. When sarcoma cells were subcutaneously injected in nude mice, tumor size, intra-tumoral microvessel density, circulating tumor cells and pulmonary metastases were significantly reduced in animals treated daily with 6 mg/Kg RI-3 as compared to animals treated with vehicle only. Thus, RI-3 represents a promising lead for anti-metastatic drugs.
Collapse
|
|
11
|
Genua M, Ingangi V, Fonteyne P, Piontini A, Yousif AM, Merlino F, Grieco P, Malesci A, Carriero MV, Danese S. Treatment with a Urokinase Receptor-derived Cyclized Peptide Improves Experimental Colitis by Preventing Monocyte Recruitment and Macrophage Polarization. Inflamm Bowel Dis 2016; 22:2390-401. [PMID: 27537052 DOI: 10.1097/mib.0000000000000896] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/09/2022]
Abstract
BACKGROUND Leukocyte migration across the blood barrier and into tissues represents a key process in the pathogenesis of inflammatory bowel diseases. The urokinase receptor (urokinase-type plasminogen activator receptor) is a master regulator of leukocyte recruitment. We recently found that cyclization of the urokinase-type plasminogen activator receptor-derived peptide Ser-Arg-Ser-Arg-Tyr [SRSRY] inhibits transendothelial migration of monocytes. Now, we have explored the effects of [SRSRY] administration during experimental colitis. METHODS The effects of [SRSRY] on cytokine profile, cytoskeletal organization, and cell migration were investigated using phorbol-12-myristate acetate-differentiated THP-1 cells exposed to polarizing stimuli. In vivo, [SRSRY] was intraperitoneally administered during dextran sodium sulfate- or 2,4,6-trinitrobenzene sulfonic acid-induced colitis in wild-type or urokinase-type plasminogen activator receptor knockout mice. Levels of pro-inflammatory cytokines and inflammatory monocytes in mucosal infiltrates were assessed by enzyme-linked immunosorbent assay and flow cytometry, respectively. RESULTS [SRSRY] prevents M0 to M1 transition and migration of M1 polarized macrophages. In vivo, [SRSRY] reduces intestinal inflammation diminishing body weight loss and disease activity index. These beneficial effects are accompanied by a reduction of interleukin 1β, interleukin 6, and tumor necrosis factor α, an increase of interleukin 10, and an abridged recruitment of inflammatory monocytes to the inflamed tissue. CONCLUSIONS Altogether, these findings indicate that [SRSRY] may be considered as a new drug useful for the pharmacological treatment of chronic inflammatory diseases, such as inflammatory bowel diseases.
Collapse
Affiliation(s)
- Marco Genua
- *IBD Center, Humanitas Research Institute, Rozzano, Italy; †Department of Translational Medicine, Università degli Studi di Milano, Milan, Italy; ‡Neoplastic Progression Unit, Department of Experimental Oncology, IRCCS Istituto Nazionale Tumori "Fondazione G. Pascale," Naples, Italy; §SUN, Second University of Naples, Naples, Italy; ‖Department of Pharmacy, University Federico II, Naples, Italy; and ¶Hunimed-Humanitas University, Milan, Italy
| | | | | | | | | | | | | | | | | | | |
Collapse
|
|
12
|
Chhokar V, Tucker AL. Angiogenesis: Basic Mechanisms and Clinical Applications. Semin Cardiothorac Vasc Anesth 2016. [DOI: 10.1177/108925320300700304] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/23/2023]
Abstract
The development and maintenance of an adequate vascular supply is critical for the viability of normal and neoplastic tissues. Angiogenesis, the development of new blood vessels from preexisting capillary networks, plays an important role in a number of physiologic and pathologic processes, including reproduction, wound repair, inflammatory diseases, and tumor growth. Angiogenesis involves sequential steps that are triggered in response to angiogenic growth factors released by inflammatory, mesenchymal, or tumor cells that act as ligands for endothelial cell receptor tyrosine kinases. Stimulated endothelial cells detach from neighboring cells and migrate, proliferate, and form tubes. The immature tubes are subsequently invested and stabilized by pericytes or smooth muscle cells. Angiogenesis depends upon complex interactions among various classes of molecules, including adhesion molecules, proteases, structural proteins, cell surface receptors, and growth factors. The therapeutic manipulation of angiogenesis targeted against ischemic and neoplastic diseases has been investigated in preclinical animal models and in clinical trials. Proangiogenic trials that have stimulated vessel growth in ischemic coronary or peripheral tissues through expression, delivery, or stimulated release of growth factors have shown efficacy in animal models and mixed results in human clinical trials. Antiangiogenic trials have used strategies to block the function of molecules critical for new vessel growth or maturation in the treatment of a variety of malignancies, mostly with results less encouraging than those seen in preclinical models. Pro-and antiangiogenic clinical trials demonstrate that strategies for optimal drug delivery, dosing schedules, patient selection, and endpoint measurements need further investigation and refinement before the therapeutic manipulation of angiogenesis will realize its full clinical potential.
Collapse
Affiliation(s)
- Vikram Chhokar
- Department of Internal Medicine, Salem VA Health System, Roanoke, Virginia
| | - Amy L. Tucker
- Department of Internal Medicine, Cardiovascular Division; Cardiovascular Research Center; Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, Virginia
| |
Collapse
|
|
13
|
Braun GB, Sugahara KN, Yu OM, Kotamraju VR, Mölder T, Lowy AM, Ruoslahti E, Teesalu T. Urokinase-controlled tumor penetrating peptide. J Control Release 2016; 232:188-95. [PMID: 27106816 PMCID: PMC5359125 DOI: 10.1016/j.jconrel.2016.04.027] [Citation(s) in RCA: 40] [Impact Index Per Article: 4.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/23/2015] [Revised: 03/30/2016] [Accepted: 04/18/2016] [Indexed: 12/11/2022]
Abstract
Tumor penetrating peptides contain a cryptic (R/K)XX(R/K) CendR element that must be C-terminally exposed to trigger neuropilin-1 (NRP-1) binding, cellular internalization and malignant tissue penetration. The specific proteases that are involved in processing of tumor penetrating peptides identified using phage display are not known. Here we design de novo a tumor-penetrating peptide based on consensus cleavage motif of urokinase-type plasminogen activator (uPA). We expressed the peptide, uCendR (RPARSGR↓SAGGSVA, ↓ shows cleavage site), on phage or coated it onto silver nanoparticles and showed that it is cleaved by uPA, and that the cleavage triggers binding to recombinant NRP-1 and to NPR-1-expressing cells. Upon systemic administration to mice bearing uPA-overexpressing breast tumors, FAM-labeled uCendR peptide and uCendR-coated nanoparticles preferentially accumulated in tumor tissue. We also show that uCendR phage internalization into cultured cancer cells and its penetration in explants of murine tumors and clinical tumor explants can be potentiated by combining the uCendR peptide with tumor-homing module, CRGDC. Our work demonstrates the feasibility of designing tumor-penetrating peptides that are activated by a specific tumor protease. As upregulation of protease expression is one of the hallmarks of cancer, and numerous tumor proteases have substrate specificities compatible with proteolytic unmasking of cryptic CendR motifs, the strategy described here may provide a generic approach for designing proteolytically-actuated peptides for tumor-penetrative payload delivery.
Collapse
Affiliation(s)
- Gary B Braun
- Sanford Burnham Prebys Medical Discovery Institute, Cancer Research Center, La Jolla, CA, USA
| | - Kazuki N Sugahara
- Sanford Burnham Prebys Medical Discovery Institute, Cancer Research Center, La Jolla, CA, USA; Department of Surgery, Columbia University College of Physicians and Surgeons, New York, NY, USA
| | - Olivia M Yu
- Sanford Burnham Prebys Medical Discovery Institute, Cancer Research Center, La Jolla, CA, USA; Biomedical Sciences Graduate Program, Department of Pharmacology, University of California San Diego, La Jolla, USA
| | | | - Tarmo Mölder
- Laboratory of Cancer Biology, Institute of Biomedicine and Translational Medicine, University of Tartu, Tartu, Estonia
| | - Andrew M Lowy
- Department of Surgery, Division of Surgical Oncology, Moores Cancer Center, University of California San Diego, La Jolla, CA, USA
| | - Erkki Ruoslahti
- Sanford Burnham Prebys Medical Discovery Institute, Cancer Research Center, La Jolla, CA, USA; Center for Nanomedicine, Department of Molecular, Cellular, and Developmental Biology, University of California, Santa Barbara, CA, USA
| | - Tambet Teesalu
- Sanford Burnham Prebys Medical Discovery Institute, Cancer Research Center, La Jolla, CA, USA; Center for Nanomedicine, Department of Molecular, Cellular, and Developmental Biology, University of California, Santa Barbara, CA, USA; Laboratory of Cancer Biology, Institute of Biomedicine and Translational Medicine, University of Tartu, Tartu, Estonia.
| |
Collapse
|
|
14
|
Vandooren J, Opdenakker G, Loadman PM, Edwards DR. Proteases in cancer drug delivery. Adv Drug Deliv Rev 2016; 97:144-55. [PMID: 26756735 DOI: 10.1016/j.addr.2015.12.020] [Citation(s) in RCA: 74] [Impact Index Per Article: 8.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/29/2015] [Revised: 12/23/2015] [Accepted: 12/24/2015] [Indexed: 01/12/2023]
Abstract
Whereas protease inhibitors have been developed successfully against hypertension and viral infections, they have failed thus far as cancer drugs. With advances in cancer profiling we now better understand that the tumor "degradome" (i.e. the repertoire of proteases and their natural inhibitors and interaction partners) forms a complex network in which specific nodes determine the global outcome of manipulation of the protease web. However, knowing which proteases are active in the tumor micro-environment, we may tackle cancers with the use of Protease-Activated Prodrugs (PAPs). Here we exemplify this concept for metallo-, cysteine and serine proteases. PAPs not only exist as small molecular adducts, containing a cleavable substrate sequence and a latent prodrug, they are presently also manufactured as various types of nanoparticles. Although the emphasis of this review is on PAPs for treatment, it is clear that protease activatable probes and nanoparticles are also powerful tools for imaging purposes, including tumor diagnosis and staging, as well as visualization of tumor imaging during microsurgical resections.
Collapse
Affiliation(s)
- Jennifer Vandooren
- KU Leuven, University of Leuven, Department of Microbiology and Immunology, Rega Institute for Medical Research, Laboratory of Immunobiology, B-3000 Leuven, Belgium
| | - Ghislain Opdenakker
- KU Leuven, University of Leuven, Department of Microbiology and Immunology, Rega Institute for Medical Research, Laboratory of Immunobiology, B-3000 Leuven, Belgium
| | - Paul M Loadman
- Institute of Cancer Therapeutics, School of Life Sciences, University of Bradford, Bradford, Yorkshire BD7 1DP, UK
| | - Dylan R Edwards
- School of Biological Sciences, University of East Anglia, Norwich Research Park, Norwich NR4 7TJ, UK; The Genome Analysis Centre, Norwich Research Park, Norwich NR4 7UH, UK.
| |
Collapse
|
|
15
|
Yousif AM, Minopoli M, Bifulco K, Ingangi V, Di Carluccio G, Merlino F, Motti ML, Grieco P, Carriero MV. Cyclization of the urokinase receptor-derived ser-arg-ser-arg-tyr Peptide generates a potent inhibitor of trans-endothelial migration of monocytes. PLoS One 2015; 10:e0126172. [PMID: 25938482 PMCID: PMC4418665 DOI: 10.1371/journal.pone.0126172] [Citation(s) in RCA: 21] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/11/2014] [Accepted: 03/30/2015] [Indexed: 02/02/2023] Open
Abstract
The receptor for the urokinase-type plasminogen activator (uPAR) is a widely recognized master regulator of cell migration and uPAR88-92 is the minimal sequence required to induce cell motility. We and others have previously documented that the uPAR88-92 sequence, even in the form of synthetic linear peptide (SRSRY), interacts with the formyl peptide receptor type 1 (FPR1), henceforth inducing cell migration of several cell lines, including monocytes. FPR1 is mainly expressed by mammalian phagocytic leukocytes and plays a crucial role in chemotaxis. In this study, we present evidence that the cyclization of the SRSRY sequence generates a new potent and stable inhibitor of monocyte trafficking. In rat basophilic leukaemia RBL-2H3/ETFR cells expressing high levels of constitutively activated FPR1, the cyclic SRSRY peptide ([SRSRY]) blocks FPR1 mediated cell migration by interfering with both internalization and ligand-uptake of FPR1. Similarly to RBL-2H3/ETFR cells, [SRSRY] competes with fMLF for binding to FPR1 and prevents agonist-induced FPR1 internalization in human monocyte THP-1 cells. Unlike scramble [RSSYR], [SRSRY] inhibits fMLF-directed migration of monocytes in a dose-dependent manner, with IC50 value of 0.01 nM. PMA-differentiated THP-1 cell exposure to fMLF gradient causes a marked cytoskeletal re-organization with the formation of F-actin rich pseudopodia that are prevented by the addition of [SRSRY]. Furthermore, [SRSRY] prevents migration of human primary monocytes and trans-endothelial migration of monocytes. Our findings indicate that [SRSRY] is a new FPR1 inhibitor which may suggest the development of new drugs for treating pathological conditions sustained by increased motility of monocytes, such as chronic inflammatory diseases.
Collapse
Affiliation(s)
| | - Michele Minopoli
- Neoplastic Progression Unit, Department of Experimental Oncology, IRCCS Istituto Nazionale Tumori “Fondazione G. Pascale”, Naples, Italy
| | - Katia Bifulco
- Neoplastic Progression Unit, Department of Experimental Oncology, IRCCS Istituto Nazionale Tumori “Fondazione G. Pascale”, Naples, Italy
| | - Vincenzo Ingangi
- Neoplastic Progression Unit, Department of Experimental Oncology, IRCCS Istituto Nazionale Tumori “Fondazione G. Pascale”, Naples, Italy
- SUN Second University of Naples, Italy
| | - Gioconda Di Carluccio
- Neoplastic Progression Unit, Department of Experimental Oncology, IRCCS Istituto Nazionale Tumori “Fondazione G. Pascale”, Naples, Italy
| | | | | | - Paolo Grieco
- Department of Pharmacy, University Federico II, Naples, Italy
- CIRPEB: Centro Interuniversitario di Ricerca sui Peptidi Bioattivi University of Naples “Federico II”, DFM-Scarl, Institute of Biostructures and Bioimaging—CNR, 80134, Naples, Italy
| | - Maria Vincenza Carriero
- Neoplastic Progression Unit, Department of Experimental Oncology, IRCCS Istituto Nazionale Tumori “Fondazione G. Pascale”, Naples, Italy
| |
Collapse
|
|
16
|
Theilade S, Lyngbaek S, Hansen TW, Eugen-Olsen J, Fenger M, Rossing P, Jeppesen JL. Soluble urokinase plasminogen activator receptor levels are elevated and associated with complications in patients with type 1 diabetes. J Intern Med 2015; 277:362-371. [PMID: 24830873 DOI: 10.1111/joim.12269] [Citation(s) in RCA: 58] [Impact Index Per Article: 5.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/25/2022]
Abstract
OBJECTIVES Soluble urokinase plasminogen activator receptor (suPAR) is a marker of inflammation and endothelial dysfunction. We investigated the associations between suPAR and diabetes, including diabetes duration and complications, in patients with type 1 diabetes. DESIGN, SETTING AND SUBJECTS From 2009 to 2011, 667 patients with type 1 diabetes and 51 nondiabetic control subjects were included in a cross-sectional study at Steno Diabetes Center, Gentofte, Denmark. suPAR levels were measured with an enzyme-linked immunosorbent assay. MAIN OUTCOME MEASURES The investigated diabetic complications were cardiovascular disease (CVD: previous myocardial infarction, revascularisation, peripheral arterial disease and stroke), autonomic dysfunction (heart rate variability during deep breathing <11 beats min(-1) ), albuminuria [urinary albumin excretion rate (UAER) ≥30 mg/24 h] or a high degree of arterial stiffness (pulse wave velocity ≥10 m s(-1) ). Analyses were adjusted for gender, age, systolic blood pressure, estimated glomerular filtration rate, UAER, glycated haemoglobin (HbA1c ), total cholesterol, body mass index, C-reactive protein, antihypertensive treatment and smoking. RESULTS Soluble urokinase plasminogen activator receptor levels were lower in control subjects versus all patients, in control subjects versus normoalbuminuric patients (UAER <30 mg/24 h), in normoalbuminuric patients with short (<10 years) versus long diabetes duration and were increased with degree of albuminuria (adjusted P < 0.001 for all). Furthermore, suPAR levels were higher in patients with versus without CVD (n = 144; 21.3%), autonomic dysfunction (n = 369; 59.2%), albuminuria (n = 357; 53.1%) and a high degree of arterial stiffness (n = 298; 47.2%) (adjusted P ≤ 0.024). The adjusted odds ratio (95% confidence interval) values per 1 ln unit increase in suPAR were as follows: 2.5 (1.1-5.7) for CVD: 2.7 (1.2-6.2) for autonomic dysfunction; 3.8 (1.3-10.9) for albuminuria and 2.5 (1.1-6.1) for a high degree of arterial stiffness (P ≤ 0.039). CONCLUSION The suPAR level is higher in patients with type 1 diabetes and is associated with diabetes duration and complications independent of other risk factors. suPAR is a potential novel risk marker for the management of diabetes.
Collapse
Affiliation(s)
- S Theilade
- Steno Diabetes Center, Gentofte, Denmark
| | - S Lyngbaek
- Department of Medicine, Glostrup Hospital, University of Copenhagen, Copenhagen, Denmark
| | - T W Hansen
- Steno Diabetes Center, Gentofte, Denmark
| | - J Eugen-Olsen
- Clinical Research Centre, Hvidovre Hospital, University of Copenhagen, Copenhagen, Denmark
| | - M Fenger
- Department of Biochemistry, Hvidovre Hospital, University of Copenhagen, Copenhagen, Denmark
| | - P Rossing
- Steno Diabetes Center, Gentofte, Denmark.,Faculty of Health, Aarhus University, Aarhus, Denmark.,Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark
| | - J L Jeppesen
- Department of Medicine, Glostrup Hospital, University of Copenhagen, Copenhagen, Denmark.,Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark
| |
Collapse
|
|
17
|
Khan R, Gupta N, Kumar R, Sharma M, Kumar L, Sharma A. Augmented expression of urokinase plasminogen activator and extracellular matrix proteins associates with multiple myeloma progression. Clin Exp Metastasis 2014; 31:585-93. [PMID: 24807734 DOI: 10.1007/s10585-014-9652-7] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/06/2013] [Accepted: 03/26/2014] [Indexed: 10/25/2022]
Abstract
Multiple myeloma (MM) represents a B cell malignancy, characterized by a monoclonal proliferation of malignant plasma cells. Interactions between tumor cells and extracellular matrix (ECM) are of importance for tumor invasion and metastasis. Protein levels of urokinase plasminogen activator (uPA) and fibulin 1, nidogen and laminin in plasma and serum respectively and mRNA levels of these molecules in peripheral blood mononuclear cells were determined in 80 subjects by using ELISA and quantitative PCR and data was analyzed with severity of disease. Pearson correlation was determined to observe interrelationship between different molecules. A statistical significant increase for ECM proteins (laminin, nidogen and fibulin 1) and uPA at circulatory level as well as at mRNA level was observed compared to healthy controls. The levels of these molecules in serum might be utilized as a marker of active disease. Significant positive correlation of all ECM proteins with uPA was found and data also correlates with severity of disease. Strong association found between ECM proteins and uPA in this study supports that there might be interplay between these molecules which can be targeted. This study on these molecules may help to gain insight into processes of growth, spread, and clinical behavior of MM.
Collapse
Affiliation(s)
- Rehan Khan
- Department of Biochemistry, All India Institute of Medical Sciences (AIIMS), New Delhi, 110029, India
| | | | | | | | | | | |
Collapse
|
|
18
|
Wyganowska-Świątkowska M, Surdacka A, Skrzypczak-Jankun E, Jankun J. The plasminogen activation system in periodontal tissue (Review). Int J Mol Med 2014; 33:763-8. [PMID: 24535478 DOI: 10.3892/ijmm.2014.1653] [Citation(s) in RCA: 16] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/17/2013] [Accepted: 01/28/2014] [Indexed: 11/05/2022] Open
Abstract
The plasminogen activation system (PAS) plays an essential role in tissue proteolysis in physiological and pathological processes. Periodontitis is a chronic infection associated with increased proteolysis driven by plasminogen activation. In this comprehensive review, we summarise the effects of PAS in wound healing, tissue remodelling, inflammation, bacterial infection, and in the initiation and progression of periodontal disease. Specifically, we discuss the role of plasminogen activators (PAs), including urokinase PA (uPA), tissue-type PA (tPA), PA inhibitor type 1 (PAI-1) and 2 (PAI-2) and activated plasminogen in periodontal tissue, where their concentrations can reach much higher values than those found in other parts of the body. We also discuss whether PA deficiencies can have effects on periodontal tissue. We conclude that in periodontal disease, PAS is unbalanced and equalizing its function can improve the clinical periodontal tissue condition.
Collapse
Affiliation(s)
| | - Anna Surdacka
- Department of Conservative Dentistry and Periodontology, Poznań University of Medical Sciences, Poznań 60-820, Poland
| | - Ewa Skrzypczak-Jankun
- Urology Research Center, Department of Urology, College of Medicine, University of Toledo, Toledo, OH 43614, USA
| | - Jerzy Jankun
- Protein Research Chair, Department of Biochemistry, College of Sciences, King Saud University, Riyadh 11451, Kingdom of Saudi Arabia
| |
Collapse
|
|
19
|
Chedraui P, Escobar GS, Pérez-López FR, Palla G, Montt-Guevara M, Cecchi E, Genazzani AR, Simoncini T. Angiogenesis, inflammation and endothelial function in postmenopausal women screened for the metabolic syndrome. Maturitas 2014; 77:370-4. [PMID: 24598235 DOI: 10.1016/j.maturitas.2014.01.014] [Citation(s) in RCA: 37] [Impact Index Per Article: 3.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/06/2014] [Revised: 01/16/2014] [Accepted: 01/27/2014] [Indexed: 01/24/2023]
Abstract
BACKGROUND Prevalence of the metabolic syndrome (METS) increases after the menopause; nevertheless, concomitant vascular, inflammatory and endothelial changes have not been completely elucidated. OBJECTIVE To measure serum markers of angiogenesis, inflammation and endothelial function in postmenopausal women screened for the METS. METHODS Serum of 100 postmenopausal women was analyzed for angiopoietin-2, interleukin-8 (IL-8), soluble FAS ligand (sFASL), interleukin-6 (IL-6), tumour necrosis factor alpha (TNF-α), soluble CD40 ligand (sCD40L), plasminogen activator inhibitor-1 (PAI-1), and urokinase-type plasminogen activator (uPA). Comparisons were made in accordance to the presence or not of the METS and each of its components. Modified Adult Treatment Panel III criteria were used to define the METS. RESULTS Women with the METS (n=57) had similar age and time since menopause as compared to those without the syndrome (n=43). In general, women with the METS displayed a trend for higher levels of the analyzed markers. Nevertheless, only IL-6 levels were found to be significantly higher and uPA levels significantly lower among METS women as compared to those without the syndrome. When analyte levels were compared as to presenting or not each of the diagnostic features of the METS, it was found that IL-6 levels were higher among women with abdominal obesity, low HDL-C and high triglyceride levels. Women with low HDL-C and high triglyceride levels presented significantly lower uPA levels and those with high glucose and low HDL-C displayed significantly higher sCD40L levels. CONCLUSION Postmenopausal women with the METS in this sample displayed higher IL-6 (inflammation) and lower uPA levels (endothelial dysfunction). These were mainly related to metabolic and lipid abnormalities. More research is warranted in this regard.
Collapse
Affiliation(s)
- Peter Chedraui
- Institute of Biomedicine, Research Area for Women's Health, Facultad de Ciencias Médicas, Universidad Católica de Santiago de Guayaquil, Guayaquil, Ecuador.
| | - Gustavo S Escobar
- Institute of Biomedicine, Research Area for Women's Health, Facultad de Ciencias Médicas, Universidad Católica de Santiago de Guayaquil, Guayaquil, Ecuador
| | - Faustino R Pérez-López
- Department of Obstetrics and Gynecology, Facultad de Medicina, Lozano Blesa University Hospital, Universidad de Zaragoza, Spain
| | - Giulia Palla
- Department of Clinical and Experimental Medicine, Division of Obstetrics and Gynecology, University of Pisa, Italy
| | - Magdalena Montt-Guevara
- Department of Clinical and Experimental Medicine, Division of Obstetrics and Gynecology, University of Pisa, Italy
| | - Elena Cecchi
- Department of Clinical and Experimental Medicine, Division of Obstetrics and Gynecology, University of Pisa, Italy
| | - Andrea R Genazzani
- Department of Clinical and Experimental Medicine, Division of Obstetrics and Gynecology, University of Pisa, Italy
| | - Tommaso Simoncini
- Department of Clinical and Experimental Medicine, Division of Obstetrics and Gynecology, University of Pisa, Italy
| | | |
Collapse
|
|
20
|
Nelis H, D'Herde K, Goossens K, Vandenberghe L, Leemans B, Forier K, Smits K, Braeckmans K, Peelman L, Van Soom A. Equine oviduct explant culture: a basic model to decipher embryo–maternal communication. Reprod Fertil Dev 2014; 26:954-66. [DOI: 10.1071/rd13089] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/14/2013] [Accepted: 06/18/2013] [Indexed: 12/27/2022] Open
Abstract
Equine embryos remain for 6 days in the oviduct and thus there is a need for an in vitro model to study embryo–oviductal interactions in the horse, since this subtle way of communication is very difficult to analyse in vivo. Until now, no equine oviduct explant culture model has been characterised both morphologically and functionally. Therefore, we established a culture system for equine oviduct explants that maintained epithelial morphology during 6 days of culture, as revealed by light microscopy and transmission electron microscopy. We demonstrated the presence of highly differentiated, tall columnar, pseudostratified epithelium with basal nuclei, numerous nucleoli, secretory granules and apical cilia, which is very similar to the in vivo situation. Both epithelium and stromal cells originating from the lamina propria are represented in the explants. Moreover, at least 98% of the cells remained membrane intact and fewer than 2% of the cells were apoptotic after 6 days of culture. Although dark-cell degeneration, which is a hypoxia-related type of cell death, was observed in the centre of the explants, quantitative real-time PCR failed to detect upregulation of the hypoxia-related marker genes HIF1A, VEGFA, uPA, GLUT1 and PAI1. Since the explants remained morphologically and functionally intact and since the system is easy to set up, it appears to be an excellent tool for proteome, transcriptome and miRNome analysis in order to unravel embryo–maternal interactions in the horse.
Collapse
|
|
21
|
Trimarchi H. Primary focal and segmental glomerulosclerosis and soluble factor urokinase-type plasminogen activator receptor. World J Nephrol 2013; 2:103-110. [PMID: 24255893 PMCID: PMC3832866 DOI: 10.5527/wjn.v2.i4.103] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 09/08/2013] [Revised: 09/24/2013] [Accepted: 10/20/2013] [Indexed: 02/06/2023] Open
Abstract
Primary focal and segmental glomerulosclerosis (FSGS) may be due to genetic or acquired etiologies and is a common cause of nephrotic syndrome with high morbidity that often leads to end-stage renal failure. The different available therapeutic approaches are unsuccessful, in part due to partially deciphered heterogeneous and complex pathophysiological mechanisms. Moreover, the term FSGS, even in its primary form, comprises a histological description shared by a number of different causes with completely different molecular pathways of disease. This review focuses on the latest developments regarding the pathophysiology of primary acquired FSGS caused by soluble factor urokinase type plasminogen activator receptor, a circulating permeability factor involved in proteinuria and edema formation, and describes recent advances with potential success in therapy.
Collapse
|
|
22
|
Chen ML, Lin YH, Yang CM, Hu ML. Lycopene inhibits angiogenesis both in vitro and in vivo by inhibiting MMP-2/uPA system through VEGFR2-mediated PI3K-Akt and ERK/p38 signaling pathways. Mol Nutr Food Res 2012; 56:889-99. [PMID: 22707264 DOI: 10.1002/mnfr.201100683] [Citation(s) in RCA: 65] [Impact Index Per Article: 5.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/20/2023]
Abstract
SCOPE Limited in vitro data show that lycopene may be anti-angiogenic but with unclear mechanisms. Here, we employed ex vivo and in vivo assays to substantiate the anti-angiogenic action of lycopene and determined its molecular mechanisms in human umbilical vein endothelial cells (HUVECs). METHODS AND RESULTS The anti-angiogenic activity of lycopene was confirmed by ex vivo rat aortic ring and in vivo chorioallantoic membrane assays. Furthermore, the in vivo matrigel plug assay in mice demonstrated that lycopene implanted s.c. at the highest dose used (400 μg/plug) completely inhibited the formation of vascular endothelial cells induced by vascular endothelial growth factor (VEGF). As expected, lycopene inhibited tube formation, invasion, and migration in HUVECs, and such actions were accompanied by reduced activities of matrix metalloproteinase-2, urokinase-type plasminogen activator, and protein expression of Rac1, and by enhancing protein expression of tissue inhibitors of metalloproteinase-2 and plasminogen activator inhibitor-1. Moreover, lycopene attenuated VEGF receptor-2 (VEGFR2)-mediated phosphorylation of extracellular signal-regulated kinase (ERK), p38, and Akt as well as protein expression of PI3K. CONCLUSION Our data demonstrate the anti-angiogenic effect of lycopene both in vitro and in vivo. The anti-angiogenic activity of lycopene may involve inhibition of MMP-2/uPA system through VEGFR2-mediated PI3K-Akt and ERK/p38 signaling pathways.
Collapse
Affiliation(s)
- Man-Ling Chen
- Department of Food Science and Biotechnology, National Chung Hsing University, Taichung, Taiwan
| | | | | | | |
Collapse
|
|
23
|
The plasminogen system in regulating stem cell mobilization. J Biomed Biotechnol 2012; 2012:437920. [PMID: 23118508 PMCID: PMC3478786 DOI: 10.1155/2012/437920] [Citation(s) in RCA: 16] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/11/2012] [Accepted: 06/05/2012] [Indexed: 12/24/2022] Open
Abstract
The treatment of patients with hematopoietic progenitor and stem cells (HPSCs) to reconstitute hematopoiesis after myeloablative therapy or to repair ischemia after myocardial infarction has significantly improved clinical outcomes. Successful blood or bone marrow transplants require a sufficient number of HPSCs capable of homing to the injured site to regenerate tissue. Granulocyte-colony stimulating factor (G-CSF) is widely used clinically for stem cell mobilization. However, in some patients the response is poor, thus a better understanding of the mechanisms underlying G-CSF-regulated stem cell mobilization is needed. The pasminogen (Plg) system is the primary fibrinolytic pathway responsible for clot dissolution after thrombosis. Recent evidence suggests that Plg plays a pivotal role in stem cell mobilization from the bone marrow to the peripheral circulation, particularly in HPSC mobilization in response to G-CSF. This paper will discuss the potential mechanisms by which the Plg system regulates stem cell mobilization, focusing on stepwise proteolysis and signal transduction during HPSC egress from their bone marrow niche. Clear elucidation of the underlying mechanisms may lead to the development of new Plg-based therapeutic strategies to improve stem cell mobilization in treating hematological and cardiovascular diseases.
Collapse
|
|
24
|
Birengel S, Yalçındağ FN, Yalçındağ A, Sahli E, Batıoğlu F. Urokinase plasminogen activator receptor levels in Behcet's disease. Thromb Res 2011; 128:274-6. [DOI: 10.1016/j.thromres.2011.03.003] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/11/2010] [Revised: 02/02/2011] [Accepted: 03/06/2011] [Indexed: 11/29/2022]
|
|
25
|
Graziano F, Elia C, Laudanna C, Poli G, Alfano M. Urokinase plasminogen activator inhibits HIV virion release from macrophage-differentiated chronically infected cells via activation of RhoA and PKCε. PLoS One 2011; 6:e23674. [PMID: 21858203 PMCID: PMC3157461 DOI: 10.1371/journal.pone.0023674] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/29/2010] [Accepted: 07/25/2011] [Indexed: 12/13/2022] Open
Abstract
BACKGROUND HIV replication in mononuclear phagocytes is a multi-step process regulated by viral and cellular proteins with the peculiar feature of virion budding and accumulation in intra-cytoplasmic vesicles. Interaction of urokinase-type plasminogen activator (uPA) with its cell surface receptor (uPAR) has been shown to favor virion accumulation in such sub-cellular compartment in primary monocyte-derived macrophages and chronically infected promonocytic U1 cells differentiated into macrophage-like cells by stimulation with phorbol myristate acetate (PMA). By adopting this latter model system, we have here investigated which intracellular signaling pathways were triggered by uPA/uPAR interaction leading the redirection of virion accumulation in intra-cytoplasmic vesicles. RESULTS uPA induced activation of RhoA, PKCδ and PKCε in PMA-differentiated U1 cells. In the same conditions, RhoA, PKCδ and PKCε modulated uPA-induced cell adhesion and polarization, whereas only RhoA and PKCε were also responsible for the redirection of virions in intracellular vesicles. Distribution of G and F actin revealed that uPA reorganized the cytoskeleton in both adherent and polarized cells. The role of G and F actin isoforms was unveiled by the use of cytochalasin D, a cell-permeable fungal toxin that prevents F actin polymerization. Receptor-independent cytoskeleton remodeling by Cytochalasin D resulted in cell adhesion, polarization and intracellular accumulation of HIV virions similar to the effects gained with uPA. CONCLUSIONS These findings illustrate the potential contribution of the uPA/uPAR system in the generation and/or maintenance of intra-cytoplasmic vesicles that actively accumulate virions, thus sustaining the presence of HIV reservoirs of macrophage origin. In addition, our observations also provide evidences that pathways controlling cytoskeleton remodeling and activation of PKCε bear relevance for the design of new antiviral strategies aimed at interfering with the partitioning of virion budding between intra-cytoplasmic vesicles and plasma membrane in infected human macrophages.
Collapse
Affiliation(s)
- Francesca Graziano
- AIDS Immunophatogenesis Unit, Division of Immunology, Transplantation and Infectious Diseases, San Raffaele Scientific Institute, Milan, Italy
| | - Chiara Elia
- AIDS Immunophatogenesis Unit, Division of Immunology, Transplantation and Infectious Diseases, San Raffaele Scientific Institute, Milan, Italy
| | - Carlo Laudanna
- Department of Pathology & Diagnostic, Faculty of Medicine and Surgery, Verona, Italy
| | - Guido Poli
- AIDS Immunophatogenesis Unit, Division of Immunology, Transplantation and Infectious Diseases, San Raffaele Scientific Institute, Milan, Italy
- Vita-Salute San Raffaele University, School of Medicine, Milan, Italy
| | - Massimo Alfano
- AIDS Immunophatogenesis Unit, Division of Immunology, Transplantation and Infectious Diseases, San Raffaele Scientific Institute, Milan, Italy
| |
Collapse
|
|
26
|
The anti-metastatic efficacy of β-ionone and the possible mechanisms of action in human hepatocarcinoma SK-Hep-1 cells. Br J Nutr 2011; 107:631-8. [PMID: 21787455 DOI: 10.1017/s0007114511003473] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/24/2022]
Abstract
β-Ionone (BI), a precursor for carotenoids, is widely distributed in fruit and vegetables. Recent in vitro studies have demonstrated the potential anti-metastatic effects of BI, but the mechanisms underlying such actions are not clear. Because liver cancer is the most endemic cancer in Taiwan and in a large region of the world, we hereby investigate the anti-metastatic effects of BI and its mechanisms of actions in a highly metastatic human hepatocarcinoma SK-Hep-1 cells. We show that incubation of cells with BI (1-50 μm) for 24 and 48 h significantly inhibited cell invasion, migration and adhesion. Mechanistically, incubation of cells with BI (1-50 μm) for 24 h resulted in the following: (1) significant inhibition of matrix metalloproteinase (MMP)-2, MMP-9 and urokinase-type plasminogen activator activities, (2) up-regulation of protein expression of the tissue inhibitor of matrix metalloproteinase (TIMP)-1, TIMP-2 and plasminogen activator inhibitor-1, (3) down-regulation of the expression of migration-related proteins, including focal adhesion kinase (FAK), phosphorylated form of FAK, Rho, Rac1 and Cdc42 and (4) up-regulation of the expression of nm23-H1 protein (P < 0·05). Overall, the results show that BI effectively inhibits the metastasis of SK-Hep-1 cells, and this effect involves the regulation of gene expression and signal pathways related to invasion and migration.
Collapse
|
|
27
|
Cheng JYC, Raghunath M, Whitelock J, Poole-Warren L. Matrix components and scaffolds for sustained islet function. TISSUE ENGINEERING PART B-REVIEWS 2011; 17:235-47. [PMID: 21476869 DOI: 10.1089/ten.teb.2011.0004] [Citation(s) in RCA: 55] [Impact Index Per Article: 3.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/29/2022]
Abstract
The clinical treatment of diabetes by islet transplantation is limited by low islet survival rates. A fundamental reason for this inefficiency is likely due to the removal of islets from their native environment. The isolation process not only disrupts interactions between blood vessels and endocrine cells, but also dramatically changes islet cell interaction with the extracellular matrix (ECM). Biomolecular cues from the ECM are important for islet survival, proliferation, and function; however, very little is known about the composition of islet ECM and the role each component plays. Without a thorough understanding of islet ECM, current endeavors to prolong islet survival via scaffold engineering lack a systematic basis. The following article reviews current knowledge of islet ECM and attempts to explain the roles they play in islet function. In addition, the effects of in vitro simulations of the native islet scaffold will be evaluated. Greater understanding in these areas will provide a preliminary platform from which a sustainable bioartificial pancreas may be developed.
Collapse
Affiliation(s)
- Jennifer Y C Cheng
- Graduate School of Biomedical Engineering, University of New South Wales, Sydney, Australia.
| | | | | | | |
Collapse
|
|
28
|
Liu X, Kolokythas A, Wang J, Huang H, Zhou X. Gene Expression Signatures of Lymph Node Metastasis in Oral Cancer: Molecular Characteristics and Clinical Significances. CURRENT CANCER THERAPY REVIEWS 2010; 6:294-307. [PMID: 21709736 PMCID: PMC3122885 DOI: 10.2174/157339410793358066] [Citation(s) in RCA: 13] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/26/2022]
Abstract
Even though lymph node metastasis accounts for the vast majority of cancer death in patients with oral cancer (OC), the molecular mechanisms of lymph node metastasis remain elusive. Genome-wide microarray analyses and functional studies in vitro and in vivo, along with detailed clinical observations, have identified a number of molecules that may contribute to lymph node metastasis. These include lymphangionenic cytokines, cell adhesion molecules, basement membrane-interacting molecules, matrix enzymes and relevant downstream signaling pathways. However, defined gene signatures from different studies are highly variable, which hinders their translation to clinically relevant applications. To date, none of the identified signatures or molecular biomarkers has been successfully implemented as a diagnostic or prognostic tool applicable to routine clinical practice. In this review, we will first introduce the significance of lymph node metastasis in OC, and clinical/experimental evidences that support the underlying molecular mechanisms. We will then provide a comprehensive review and integrative analysis of the existing gene expression studies that aim to identify the metastasis-related signatures in OC. Finally, the remaining challenges will be discussed and our insights on future directions will be provided.
Collapse
Affiliation(s)
- Xiqiang Liu
- Center for Molecular Biology of Oral Diseases, College of Dentistry, University of Illinois at Chicago, Chicago, IL
- Research Institute & the Affiliated Hospital of Stomatology, Sun Yat-Sen University, Guangzhou, China
| | - Antonia Kolokythas
- Department of Oral and Maxillofacial Surgery, College of Dentistry, University of Illinois at Chicago, Chicago, IL
| | - Jianguang Wang
- Department of Oral and Maxillofacial Surgery, the Second Affiliated Hospital, Sun Yat-Sen University, Guangzhou, China
| | - Hongzhang Huang
- Research Institute & the Affiliated Hospital of Stomatology, Sun Yat-Sen University, Guangzhou, China
| | - Xiaofeng Zhou
- Center for Molecular Biology of Oral Diseases, College of Dentistry, University of Illinois at Chicago, Chicago, IL
- Research Institute & the Affiliated Hospital of Stomatology, Sun Yat-Sen University, Guangzhou, China
- Graduate College, and UIC Cancer Center, University of Illinois at Chicago, Chicago, IL
| |
Collapse
|
|
29
|
Garg N, Goyal N, Strawn TL, Wu J, Mann KM, Lawrence DA, Fay WP. Plasminogen activator inhibitor-1 and vitronectin expression level and stoichiometry regulate vascular smooth muscle cell migration through physiological collagen matrices. J Thromb Haemost 2010; 8:1847-54. [PMID: 20492459 PMCID: PMC2941703 DOI: 10.1111/j.1538-7836.2010.03907.x] [Citation(s) in RCA: 31] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/09/2023]
Abstract
BACKGROUND Vascular smooth muscle cell (VSMC) migration is a critical process in arterial remodeling. Purified plasminogen activator inhibitor-1 (PAI-1) is reported to both promote and inhibit VSMC migration on two-dimensional (D) surfaces. OBJECTIVE To determine the effects of PAI-1 and vitronectin (VN) expressed by VSMC themselves on migration through physiological collagen matrices. METHODS We studied migration of wild-type (WT), PAI-1-deficient, VN-deficient, PAI-1/VN doubly-deficient (DKO) and PAI-1-transgenic (Tg) VSMC through three-D collagen gels. RESULTS WT VSMC migrated significantly slower than PAI-1- and VN-deficient VSMC, but significantly faster than DKO VSMC. Experiments with recombinant PAI-1 suggested that basal VSMC PAI-1 expression inhibits migration by binding VN, which is secreted by VSMC and binds collagen. However, PAI-1-over-expressing Tg VSMC migrated faster than WT VSMC. Reconstitution experiments with recombinant PAI-1 mutants suggested that the pro-migratory effect of PAI-1 over-expression required its anti-plasminogen activator (PA) and LDL receptor-related protein (LRP) binding functions, but not VN binding. While promoting VSMC migration in the absence of PAI-1, VN inhibited the pro-migratory effect of active PAI-1. CONCLUSIONS In isolation, VN and PAI-1 are each pro-migratory. However, via formation of a high-affinity, non-motogenic complex, PAI-1 and VN each buffers the other's pro-migratory effect. The level of PAI-1 expression by VSMC and the concentration of VN in extracellular matrix are critical determinants of whether PAI-1 and VN promote or inhibit migration. These findings help to rectify previously conflicting reports and suggest that PAI-1/VN stoichiometry plays an important role in VSMC migration and vascular remodeling.
Collapse
Affiliation(s)
- N Garg
- Department of Internal Medicine, University of Missouri School of Medicine and Research Service, Harry S. Truman Memorial Veterans Affairs Hospital, Columbia, MO, USA
| | | | | | | | | | | | | |
Collapse
|
|
30
|
Srikanchai T, Murani E, Wimmers K, Ponsuksili S. Four loci differentially expressed in muscle tissue depending on water-holding capacity are associated with meat quality in commercial pig herds. Mol Biol Rep 2010; 37:595-601. [PMID: 19823956 DOI: 10.1007/s11033-009-9856-0] [Citation(s) in RCA: 17] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/22/2009] [Accepted: 09/28/2009] [Indexed: 11/28/2022]
Abstract
Four genes, VTN, KERA, LYZ, and a non-annotated EST (Affymetrix probe set ID: Ssc.25503.1.S1_at), whose candidacy for traits related to water-holding capacity of meat arises from their trait-dependent differential expression, were selected for candidate gene analysis. Based on in silico analysis SNPs were detected, confirmed by sequencing and used to genotype animals of 4 pig populations including 3 commercial herds of Pietrain (PI), Pietrain x (German Large White x German Landrace) (PIF1), German Landrace (DL) and 1 experimental F2 population Duroc x Pietrain (DUPI). Comparative and genetic mapping established the location of VTN on SSC12, of LYZ and KERA on SSC5 and of UN on SSC7, coinciding with QTL regions for meat quality traits. VTN showed association with pH1, pH24 and drip loss. LYZ revealed association with conductivity 24, pH1 and drip loss. KERA was associated with pH. UN showed association with pH24 and drip loss, respectively. However, none of the candidate genes showed significant associations for a particular trait across all populations. This may be due to breed specific effects that are related to the differences in meat quality of theses pig breeds. The studies revealed statistic evidence for a link of genetic variation at these loci or close to them and promoted those four candidate genes as functional and/or positional candidate genes for meat quality traits.
Collapse
Affiliation(s)
- Tiranun Srikanchai
- Research Institute for the Biology of Farm Animals, Dummerstorf, Germany
| | | | | | | |
Collapse
|
|
31
|
Abstract
Rouget, in 1873, was the first to describe a population of cells surrounding capillaries, which he regarded as contractile elements. Fifty years later, Zimmermann termed these cells "pericytes" and distinguished three subtypes along the vascular tree. Since then, the discussion concerning the contractile ability of pericytes has never ceased. Current concepts of pericyte biology rather suggest critical roles in the maintenance of homeostasis, blood-brain barrier (BBB) integrity, angiogenesis, and neovascularization. In addition, data from models of brain pathology suggest that novel pericytes are recruited from the bone marrow, but their respective precursor remains enigmatic. Recent data also suggest an important role in the regulation of cerebral blood flow, thus confirming Rouget's original idea. However, comparison of data from different studies is often constrained by the fact that pericytes were questionably identified. Although a clear-cut definition exists, defining pericytes as part of the vascular wall being enclosed in its basement membrane, pericytes are often mixed up with adjacent cell types of the vascular wall, the perivascular space, and the juxtavascular parenchyma. In fact, their identification is difficult-if not impossible-in standard histological sections. An unambiguous distinction, however, is possible at the ultrastructural level and in semi-thin sections, where their location within the vascular basement membrane can be displayed. Using these techniques in combination with immunological staining methods allows demarking their unique morphology and location. Here, we review original papers describing pericytes, briefly outline their topography within the vascular compartments, describe methods for their identification, and summarize current concepts of their function.
Collapse
Affiliation(s)
- Martin Krueger
- Dr. Senckenbergische Anatomie, Institute of Clinical Neuroanatomy, J W Goethe-University, Frankfurt/Main, Germany
| | | |
Collapse
|
|
32
|
Stewart CE, Hall IP, Parker SG, Moffat MF, Wardlaw AJ, Connolly MJ, Ruse C, Sayers I. PLAUR polymorphisms and lung function in UK smokers. BMC MEDICAL GENETICS 2009; 10:112. [PMID: 19878584 PMCID: PMC2784766 DOI: 10.1186/1471-2350-10-112] [Citation(s) in RCA: 25] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 06/03/2009] [Accepted: 10/31/2009] [Indexed: 12/15/2022]
Abstract
Background We have previously identified Urokinase Plasminogen Activator Receptor (PLAUR) as an asthma susceptibility gene. In the current study we tested the hypothesis that PLAUR single nucleotide polymorphisms (SNPs) determine baseline lung function and contribute to the development of Chronic Obstructive Pulmonary Disease (COPD) in smokers. Methods 25 PLAUR SNPs were genotyped in COPD subjects and individuals with smoking history (n = 992). Linear regression was used to determine the effects of polymorphism on baseline lung function (FEV1, FEV1/FVC) in all smokers. Genotype frequencies were compared in spirometry defined smoking controls (n = 176) versus COPD cases (n = 599) and COPD severity (GOLD stratification) using logistic regression. Results Five SNPs showed a significant association (p < 0.01) with baseline lung function; rs2302524(Lys220Arg) and rs2283628(intron 3) were associated with lower and higher FEV1 respectively. rs740587(-22346), rs11668247(-20040) and rs344779(-3666) in the 5'region were associated with increased FEV1/FVC ratio. rs740587 was also protective for COPD susceptibility and rs11668247 was protective for COPD severity although no allele dose relationship was apparent. Interestingly, several of these associations were driven by male smokers not females. Conclusion This study provides tentative evidence that the asthma associated gene PLAUR also influences baseline lung function in smokers. However the case-control analyses do not support the conclusion that PLAUR is a major COPD susceptibility gene in smokers. PLAUR is a key serine protease receptor involved in the generation of plasmin and has been implicated in airway remodelling.
Collapse
Affiliation(s)
- Ceri E Stewart
- Division of Therapeutics & Molecular Medicine, Nottingham Respiratory Biomedical Research Unit, University Hospital of Nottingham, Nottingham, UK.
| | | | | | | | | | | | | | | |
Collapse
|
|
33
|
TNFα-mediated plasminogen activation on neutrophils is involved in the high plasmin activity in mammary secretion of drying-off cows. J DAIRY RES 2009; 76:459-68. [DOI: 10.1017/s0022029909990069] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/06/2022]
Abstract
Interactions between inflammatory cytokines and plasminogen (Pg) activation system on immune cells are yet to be established. In previous studies we reported a somatic cell-associated elevation of proteolytic activity in mammary secretion of drying-off goats and cows. The purposes of the present study were to examine the role of TNF-α in polymorphonuclear neutrophil (PMN)-associated Pg activation, and the significance of this activation pathway for overall plasmin (Pm) activity in mammary secretion of drying-off cows. Results of experiments in vitro showed that the spontaneous Pg activation observed on fresh preparations of bovine blood PMN was completely blocked by anti bovine TNF-α antibody, and was further up-regulated by exogenous bovine TNF-α. Monitoring the parameters of mammary secretion of drying-off cows revealed that both somatic cell counts and differential PMN ratio was significantly elevated at weeks 1, 2 and 3 of milk stasis. Nevertheless, specific activity of soluble Pm in mammary secretion increased and the level of 17-kDa TNF-α decreased immediately following milk stasis. Iimmunoblotting revealed that although both 26-kDa pro-TNF-α and 17-kDa TNF-α were consistently present in somatic cells of mammary secretion collected at weeks 0, 1, 2 and 3 of milk stasis, only 26-kDa pro-TNF-α was present in somatic cells of milk during lactation. In-vitro assay indicated that cell-free mammary secretion of drying-off cows exerted no Pg activation bioactivity towards bovine blood PMN. Altogether, the current study suggests the existence of an active TNF-α-Pg-Pm autocrine/paracrine loop on the massively infiltrated PMN inside udders of drying-off cows, which involves extensive binding and internalization of 17-kDa TNF-α on PMN and consequently activation of Pg, resulting in high Pm activity and low 17-kDa TNF-α level in mammary secretion. These coordinated mechanisms may play a role in the defence of drying-off mammary gland.
Collapse
|
|
34
|
Esselens CW, Malapeira J, Colomé N, Moss M, Canals F, Arribas J. Metastasis-associated C4.4A, a GPI-anchored protein cleaved by ADAM10 and ADAM17. Biol Chem 2008; 389:1075-84. [PMID: 18979631 DOI: 10.1515/bc.2008.121] [Citation(s) in RCA: 28] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/15/2022]
Abstract
Metalloproteases play a complex role in tumor progression. While the activity of some ADAM, ADAMTS and matrix metalloproteases (MMPs) seems to be protumorigenic, the activity of others seems to prevent tumor progression. The identification of the array of substrates of a given metalloprotease (degradome) seems an adequate approach to predict the effect of the inhibition of a metalloprotease in tumors. Here, we present the proteomic identification of a novel substrate for ADAM10 and -17. We used SILAC (Stable Isotope Labeling by Amino acids in Cell culture), a proteomic technique based on the differential metabolic labeling of cells in different conditions. This was applied to MCF7 cells derived from an invasive mammary tumor, and the same cells expressing shRNAs that knock down ADAM10 or -17. Following this approach, we have identified C4.4A as a substrate to both metalloproteases. Since C4.4A is likely involved in tumor invasion, these results indicate that the cleavage of C4.4A by ADAM10 and ADAM17 contributes to tumor progression.
Collapse
Affiliation(s)
- Cary W Esselens
- Medical Oncology Research Program, Vail d'Hebron University Hospital Research Institute, Psg. Vail d'Hebron 119-129, Universitat Autonoma de Barcelona, E-08035 Barcelona, Spain
| | | | | | | | | | | |
Collapse
|
|
35
|
Ligand-engaged urokinase-type plasminogen activator receptor and activation of the CD11b/CD18 integrin inhibit late events of HIV expression in monocytic cells. Blood 2008; 113:1699-709. [PMID: 18941116 DOI: 10.1182/blood-2008-02-138412] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/22/2022] Open
Abstract
Urokinase-type plasminogen activator (uPA) signaling via its receptor uPAR inhibits late events in HIV-1 replication in acutely infected primary monocyte-derived macrophages (MDMs) and promonocytic U937 cells. Here we show that U937-derived, chronically infected U1 cells stimulated with phorbol 12-myristate 13-acetate (PMA) express integrins, uPA, and soluble uPAR at levels similar to those of MDMs. uPA inhibited HIV expression in U1 cells incubated with either PMA or tumor necrosis factor-alpha (TNF-alpha), but not with other HIV-inductive cytokines or lipopolysaccharide. Of interest, only PMA and TNF-alpha, but not other HIV-inductive stimuli, induced surface expression of the alpha(M) chain CD11b in U1 cells constitutively expressing CD18, the beta(2) chain of the Mac-1 integrin. Like uPA, fibrinogen, a Mac-1 (CD11b/CD18) ligand, and M25, a peptide homologous to a portion of the beta-propeller region of CD11b preventing its association with uPAR, inhibited HIV virion release in PMA-stimulated U1 cells. Both uPAR small-interference RNA (siRNA) and soluble anti-beta(1)/-beta(2) monoclonal antibodies abolished the anti-HIV effects of uPA, whereas CD11b siRNA reversed the anti-HIV effect of M25, but not that induced by uPA. Thus, either uPA/uPAR interaction, Mac-1 activation, or prevention of its association with uPAR triggers a signaling pathway leading to the inefficient release of HIV from monocytic cells.
Collapse
|
|
36
|
The urokinase system in patients with intermittent and persistent allergic rhinitis. Blood Coagul Fibrinolysis 2008; 19:685-8. [DOI: 10.1097/mbc.0b013e32830b287d] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/26/2022]
|
|
37
|
Esselens CW, Malapeira J, Colomé N, Moss M, Canals F, Arribas J. Metastasis-associated C4.4A, a GPI-anchored protein cleaved by ADAM10 and ADAM17. Biol Chem 2008. [DOI: 10.1515/bc.2008.121_bchm.just-accepted] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/15/2022]
|
|
38
|
Bacchiocchi R, Rubini C, Pierpaoli E, Borghetti G, Procacci P, Nocini PF, Santarelli A, Rocchetti R, Ciavarella D, Lo Muzio L, Fazioli F. Prognostic value analysis of urokinase-type plasminogen activator receptor in oral squamous cell carcinoma: an immunohistochemical study. BMC Cancer 2008; 8:220. [PMID: 18673553 PMCID: PMC2527016 DOI: 10.1186/1471-2407-8-220] [Citation(s) in RCA: 25] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/06/2008] [Accepted: 08/01/2008] [Indexed: 11/30/2022] Open
Abstract
Background Oral squamous cell carcinoma (OSCC) represents the most common oral malignancy. Despite recent advances in therapy, up to 50% of the cases have relapse and/or metastasis. There is therefore a strong need for the identification of new biological markers able to predict the clinical behaviour of these lesions in order to improve quality of life and overall survival. Among tumour progression biomarkers, already known for their involvement in other neoplasia, a crucial role is ascribed to the urokinase-type plasminogen activator receptor (uPAR), which plays a multiple role in extracellular proteolysis, cell migration and tissue remodelling not only as a receptor for the zymogen pro-uPA but also as a component for cell adhesion and as a chemoattractant. The purpose of this study was to gain information on the expression of uPAR in OSCC and to verify whether this molecule can have a role as a prognostic/predictive marker for this neoplasia. Methods In a retrospective study, a cohort of 189 OSCC patients was investigated for uPAR expression and its cellular localization by immunohistochemistry. As standard controls, 8 normal oral mucosal tissues free of malignancy, obtained from patients with no evidence or history of oral cavity tumours, were similarly investigated. After grouping for uPAR expression, OSCCs were statistically analyzed for the variables age, gender, histological grading (G), tumour size, recurrence, TNM staging and overall survival rate. Results In our immunohistochemical study, 74 cases (39.1%) of OSCC showed a mostly cytoplasmic positivity for uPAR, whereas 115 were negative. uPAR expression correlated with tumour differentiation grade and prognosis: percentage of positive cases was the greatest in G3 (70.4%) and patients positives for uPAR expression had an expectation of life lower than those for uPAR negatives. Conclusion The results obtained in this study suggest a role of uPAR as a potential biomarker useful to identify higher risk subgroups of OSCC patients.
Collapse
Affiliation(s)
- Roberta Bacchiocchi
- Department of Molecular Pathology and Innovative Therapies, Polytechnic University of the Marche Region, Ancona, Italy.
| | | | | | | | | | | | | | | | | | | | | |
Collapse
|
|
39
|
Csutak A, Silver DM, Tozsér J, Steiber Z, Bagossi P, Hassan Z, Berta A. Plasminogen activator inhibitor in human tears after laser refractive surgery. J Cataract Refract Surg 2008; 34:897-901. [PMID: 18498992 DOI: 10.1016/j.jcrs.2008.02.024] [Citation(s) in RCA: 16] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/07/2007] [Accepted: 02/27/2008] [Indexed: 11/15/2022]
Abstract
PURPOSE To observe levels of plasminogen activator inhibitor (PAI) in human tears after photorefractive keratectomy (PRK) and laser in situ keratomileusis (LASIK). SETTING University medical center eye clinic. METHODS Tear samples were collected from 46 eyes having PRK and 13 eyes having LASIK immediately before and after surgery and on the first (LASIK), third (PRK), and fifth (PRK) postoperative days. Analyses used enzyme-linked immunoassay, yielding 61 PRK PAI-1 determinations and 146 PRK and 35 LASIK PAI-2 determinations. RESULTS All determinations of PRK PAI-1 were below the detection limit of 1 ng/mL of the original tear sample. In the PRK eyes, the mean PAI-2 concentration was 19.8 ng/mL +/- 23.4 (SD) in preoperative tears, 112.7 +/- 60.5 ng/mL immediately postoperatively, 12.1 +/- 19.5 ng/mL after 3 days, and 15.5 +/- 20.4 ng/mL after 5 days. In the LASIK eyes, the mean PAI-2 concentration was 19.0 +/- 33.1 ng/mL preoperatively, 111.5 +/- 69.2 ng/mL immediately postoperatively, and 15.7 +/- 18.8 ng/mL after 1 day. CONCLUSIONS The similarity in the general time pattern of PAI-2 after PRK and LASIK suggests commonality in the enzymatic control response to corneal surgical wounding. Taken in the context of previous work, the observed levels of PAI-2 concentration in eyes with and without opacification suggest that in the postsurgical period, PAI-2 is not the controlling mechanism for the later development of corneal opacification and haze.
Collapse
Affiliation(s)
- Adrienne Csutak
- Department of Ophthalmology, University of Debrecen, Medical and Health Science Center, Faculty of Medicine, Debrecen, Hungary
| | | | | | | | | | | | | |
Collapse
|
|
40
|
Chen SC, Henry DO, Reczek PR, Wong MK. Plasminogen activator inhibitor-1 inhibits prostate tumor growth through endothelial apoptosis. Mol Cancer Ther 2008; 7:1227-36. [DOI: 10.1158/1535-7163.mct-08-0051] [Citation(s) in RCA: 25] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/16/2022]
|
|
41
|
Qian HS, Gu JM, Liu P, Kauser K, Halks-Miller M, Vergona R, Sullivan ME, Dole WP, Deng GG. Overexpression of PAI-1 prevents the development of abdominal aortic aneurysm in mice. Gene Ther 2007; 15:224-32. [DOI: 10.1038/sj.gt.3303069] [Citation(s) in RCA: 38] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/09/2022]
|
|
42
|
Induction of Indefinite Cardiac Allograft Survival Correlates With Toll-Like Receptor 2 and 4 Downregulation After Serine Protease Inhibitor-1 (Serp-1) Treatment. Transplantation 2007; 84:1158-67. [DOI: 10.1097/01.tp.0000286099.50532.b0] [Citation(s) in RCA: 24] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/26/2022]
|
|
43
|
Elia C, Cassol E, Sidenius N, Blasi F, Castagna A, Poli G, Alfano M. Inhibition of HIV replication by the plasminogen activator is dependent on vitronectin-mediated cell adhesion. J Leukoc Biol 2007; 82:1212-20. [PMID: 17704294 DOI: 10.1189/jlb.0407251] [Citation(s) in RCA: 14] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/24/2022] Open
Abstract
Urokinase-type plasminogen activator (uPA), an inducer of macrophage adhesion, inhibits HIV-1 expression in PMA-stimulated, chronically infected U1 cells. We investigated whether uPA-dependent cell adhesion played a role in uPA-dependent inhibition of HIV-1 replication in these cells. Monocyte-derived macrophages (MDM) were generated from monocytes of HIV-infected individuals or from cells of seronegative donors infected acutely in vitro. U1 cells were stimulated in the presence or absence of uPA in standard tissue culture (TC) plates, allowing firm cell adhesion or ultra-low adhesion (ULA) plates. Moreover, U1 cells were also maintained in the presence or absence of vitronectin (VN)-containing sera or serum from VN(-/-) mice. Virus production was evaluated by RT activity in culture supernatants, whereas cell adhesion was by crystal violet staining and optical microscopy. uPA inhibited HIV replication in MDM and PMA-stimulated U1 cells in TC plates but not in ULA plates. uPA failed to inhibit HIV expression in U1 cells stimulated with IL-6, which induces virus expression but not cell adhesion in TC plates. VN, known to bind to the uPA/uPA receptor complex, was crucial for these adhesion-dependent, inhibitory effects of uPA on HIV expression, in that they were not observed in TC plates in the presence of VN(-/-) mouse serum. HIV production in control cell cultures was increased significantly in ULA versus TC plates, indicating that macrophage cell adhesion per se curtails HIV replication. In conclusion, uPA inhibits HIV-1 replication in macrophages via up-regulation of cell adhesion to the substrate mediated by VN.
Collapse
Affiliation(s)
- Chiara Elia
- DIBIT, AIDS Immunopathogenesis Unit, San Raffaele Scientific Institute, Via Olgettina, 58, 20132, Milan, Italy
| | | | | | | | | | | | | |
Collapse
|
|
44
|
Abstract
The N-terminal somatomedin B domain (SMB) of vitronectin binds PAI-1 and the urokinase receptor with high affinity and regulates tumor cell adhesion and migration. We have shown previously in the crystal structure of the PAI-1/SMB complex that SMB, a peptide of 51 residues, is folded as a compact cysteine knot of four pairs of crossed disulfide bonds. However, the physiological significance of this structure was questioned by other groups, who disputed the disulfide bonding shown in the crystal structure (Cys5-Cys21, Cys9-Cys39, Cys19-Cys32, Cys25-Cys31), notably claiming that the first disulfide is Cys5-Cys9 rather than the Cys5-Cys21 bonding shown in the structure. To test if the claimed Cys5-Cys9 bond does exist in the SMB domain of plasma vitronectin, we purified mouse and rat plasma vitronectin that have a Met (hence cleavable by cyanogen bromide) at residue 14, and also prepared recombinant human SMB variants from insect cells with residues Asn14 or Leu24 mutated to Met. HPLC and mass spectrometry analysis showed that, after cyanogen bromide digestion, all the fragments of the SMB derived from mouse or rat vitronectin or the recombinant SMB mutants are still linked together by disulfides, and the N-terminal peptide (residue 1-14 or 1-24) can only be released when the disulfide bonds are broken. This clearly demonstrates that Cys5 and Cys9 of SMB do not form a disulfide bond in vivo, and together with other structural evidence confirms that the only functional structure of the SMB domain of plasma vitronectin is that seen in its crystallographic complex with PAI-1.
Collapse
Affiliation(s)
- Aiwu Zhou
- Department of Haematology, Cambridge Institute for Medical Research, University of Cambridge, UK.
| |
Collapse
|
|
45
|
Wang H, Zhang Y, Heuckeroth RO. Tissue-type plasminogen activator deficiency exacerbates cholestatic liver injury in mice. Hepatology 2007; 45:1527-37. [PMID: 17538930 DOI: 10.1002/hep.21613] [Citation(s) in RCA: 26] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 01/13/2023]
Abstract
UNLABELLED Recent studies demonstrating a role for plasminogen activator inhibitor (PAI)-1 in cholestatic liver disease in mice suggested that tissue-type plasminogen activator (tPA) or urokinase plasminogen activator (uPA) might be important after biliary tract obstruction. We now demonstrate that blocking tPA exacerbates liver injury after bile duct ligation (BDL). tPA deficient mice have increased bile infarcts, increased TUNEL positive cells, increased neutrophil infiltration, reduced hepatocyte proliferation and reduced ductular reaction 72 hours after BDL compared to wild type mice. In addition, the protective and proliferative effects of plasminogen activator inhibitor 1 (PAI-1) deficiency after BDL are dramatically blocked by the tPA inhibitor tPA-STOP. One potential mechanism for these effects is that both tPA deficiency and tPA-STOP reduce hepatocyte growth factor (HGF) activation and c-Met phosphorylation in the liver after BDL. In support of this hypothesis, HGF treatment reverses the effects of tPA deficiency in mice. Furthermore, preferential tPA activation in areas of injury after BDL might occur because fibrin accumulates in bile infarcts and activates tPA. CONCLUSION tPA inactivation accelerates liver injury after BDL and reduces HGF activation. These data suggest that strategies to increase HGF activation might be protective in liver diseases with biliary tract obstruction even without increased HGF production.
Collapse
Affiliation(s)
- Hongtao Wang
- Division of Gastroenterology and Nutrition, Department of Pediatrics, Washington University School of Medicine, 660 S. Euclid Avenue, St. Louis, MO 63110, USA
| | | | | |
Collapse
|
|
46
|
Lee J, Duk Jung I, Gyo Park C, Han JW, Young Lee H. Autotaxin stimulates urokinase-type plasminogen activator expression through phosphoinositide 3-kinase-Akt-nuclear [corrected] factor kappa B signaling cascade in human melanoma cells. Melanoma Res 2007; 16:445-52. [PMID: 17013094 DOI: 10.1097/01.cmr.0000232293.14408.a4] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/28/2023]
Abstract
Autotaxin, a lysophospholipase D producing lysophosphatidic acid, augments invasive and metastatic potential of tumor cells. Current investigations have focused on understanding the molecular mechanisms by which autotaxin regulates the expression of a major mediator of tumor invasion and metastasis, urokinase-type plasminogen activator (uPA) in human A2058 melanoma cells. Autotaxin induced uPA expression in a dose-dependent manner that was inhibited by pharmacological inhibitors for Gi (pertussis toxin), phosphoinositide 3-kinase (PI3K, LY294002), Akt inhibitor (AktI), proteosome activity and IkappaB phosphorylation (pyrrolidine dithiocarbamate), and by a dominant negative mutant (DN) of Akt. Autotaxin phosphorylated Akt and induced the translocation of nuclear [corrected] factor-kappaB (NF-kappaB) to the nucleus that were inhibited by AktI or by overexpressing DN-Akt. Consistently, green fluorescence protein-tagged p65 of NF-kappaB accumulated in the nucleus by autotaxin that was abrogated when the cells were transfected with DN-Akt. Moreover, autotaxin increased the DNA binding ability of NF-kappaB and promoter activity of uPA. Collectively, these data strongly suggest autotaxin induces uPA expression via the Gi-PI3K-Akt-NF-kappaB signaling pathway that might be critical for autotaxin-induced tumor cell invasion and metastasis.
Collapse
Affiliation(s)
- Jangsoon Lee
- College of Medicine, Konyang University, Daejeon, Korea
| | | | | | | | | |
Collapse
|
|
47
|
Wang XQ, Bdeir K, Yarovoi S, Cines DB, Fang W, Abraham E. Involvement of the urokinase kringle domain in lipopolysaccharide-induced acute lung injury. THE JOURNAL OF IMMUNOLOGY 2007; 177:5550-7. [PMID: 17015742 DOI: 10.4049/jimmunol.177.8.5550] [Citation(s) in RCA: 30] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 11/19/2022]
Abstract
Urokinase plasminogen activator (uPA) plays a major role in fibrinolytic processes and also can potentiate LPS-induced neutrophil activation through interactions with its kringle domain (KD). To investigate the role of the uPA KD in modulating acute inflammatory processes in vivo, we cloned and then developed Abs to the murine uPA KD. Increased pulmonary expression of uPA and the uPA KD was present in the lungs after LPS exposure. Administration of anti-kringle Abs diminished LPS-induced up-regulation of uPA and uPA KD in the lungs, and also decreased the severity of LPS-induced acute lung injury, as determined by development of lung edema, pulmonary neutrophil accumulation, histology, and lung IL-6, MIP-2, and TNF-alpha cytokine levels. These proinflammatory effects of the uPA KD appeared to be mediated through activation of Akt and NF-kappaB. The present studies indicate that the uPA KD plays a major role in the development of TLR4-mediated acute inflammatory processes, including lung injury. Blockade of the uPA KD may prevent the development or ameliorate the severity of acute lung injury induced through TLR4-dependent mechanisms, such as would occur in the setting of Gram-negative pulmonary or systemic infection.
Collapse
Affiliation(s)
- Xue-Qing Wang
- Division of Pulmonary Sciences and Critical Care Medicine, University of Colorado, and Health Services Center, Denver, CO 80262, USA
| | | | | | | | | | | |
Collapse
|
|
48
|
Kim CK, Hong SH, Joe YA, Shim BS, Lee SK, Hong YK. The recombinant kringle domain of urokinase plasminogen activator inhibits in vivo malignant glioma growth. Cancer Sci 2007; 98:253-8. [PMID: 17233842 PMCID: PMC11158234 DOI: 10.1111/j.1349-7006.2006.00378.x] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/30/2022] Open
Abstract
In a previous report, the recombinant kringle domain (UK1) of the urokinase type plasminogen activator (uPA) showed antiangiogenic activity. Here, we investigated in vivo antitumor effects of the UK1 of human uPA employing a brain tumor model. The systemic administration of UK1 purified from pichia expression (10 and 50 mg/kg/day intraperitoneally for 25 days) led to suppress the growth of a U87 human glioma xenograft, implanted into the brains of male BALB/cSlc nude mice, by 35% and 80%, respectively. In the immunohistochemical analysis, the tumors treated with UK1 showed decreased vascularity and expression of angiogenesis-related factors including vascular endothelial growth factor (VEGF), angiogenin, alpha-smooth muscle actin, von Willebrand's factor, and CD31 (PECAM-1 [Platelet endothelial cell adhesion molecule-1]), and increased apoptosis. UKl inhibited the in vitro proliferation and tube formation of VEGF-stimulated endothelial cells but not the proliferation of glioma cells. These results suggest that UK1 inhibits the malignant glioma growth by suppression of angiogenesis.
Collapse
Affiliation(s)
- Chung Kwon Kim
- Cancer Research Institute, Catholic University of Korea, Seoul 137-701, Republic of Korea
| | | | | | | | | | | |
Collapse
|
|
49
|
Kasperska-Zajac A, Rogala B. Circulating levels of urokinase-type plasminogen activator (uPA) and its soluble receptor (suPAR) in patients with atopic eczema/dermatitis syndrome. Inflammation 2007; 29:90-3. [PMID: 16858643 DOI: 10.1007/s10753-006-9004-0] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/24/2022]
Abstract
Accumulating evidence has indicated that different immune and inflammatory processes may be accompanied by up-regulation of the uPA/uPAR system. Moreover, it has been suggested that the fibrinolytic system participates actively in immune-mediated skin disorders, including atopic eczema/dermatitis syndrome (AEDS). To study a possible role of such uPA/uPAR system in AEDS, we investigated circulating levels of uPA and suPAR in patients at different clinical stages of AEDS. Levels of u-PA and suPAR were measured by enzyme-linked immunoassay in plasma from 13 patients (five females and eight males; median age 27 years) with moderate AEDS, eight patients (three females and five males; median age 25.5 years) with severe AEDS, and 18 age- and sex-matched healthy subjects. Plasma levels of uPA and suPAR in AEDS patients did not differ significantly when compared with those in healthy subjects. Moreover, we failed to observe any significant differences in levels of these components between patients with moderate and severe AEDS and the controls. It seems that plasma levels of uPA and suPAR are similar in patients at the different stages of AEDS and the healthy subjects. Moreover, these data suggest that the release of uPA and its soluble receptor into the bloodstream is not increased in the course of complex immune-mediated processes associated with AEDS.
Collapse
Affiliation(s)
- Alicja Kasperska-Zajac
- Chair and Clinical Department of Internal Diseases, Allergology and Clinical Immunology, Medical University of Silesia, Katowice, Poland.
| | | |
Collapse
|
|
50
|
Kasperska-Zajac A, Brzoza Z, Rogala B. Blood urokinase plasminogen activator system in chronic urticaria. Arch Dermatol Res 2006; 298:409-11. [PMID: 17102952 DOI: 10.1007/s00403-006-0715-8] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/03/2006] [Revised: 09/26/2006] [Accepted: 10/13/2006] [Indexed: 11/27/2022]
Abstract
The evidence gathered has pointed to the fibrinolytic system, which apart from its major role in hemostasis may also be involved in inflammatory and immune processes. To understand better the role of fibrinolysis in urticaria, we measured plasma levels of the urokinase system associated molecules such as urokinase-type plasminogen activator (uPA), its soluble receptor (suPAR; CD87) and an inhibitor, plasminogen activator inhibitor type 1 (PAI-1) activity in chronic urticaria (CU) patients. Plasma was obtained from symptomatic sixteen CU patients (12 females and 4 males) showing positive response to autologous serum skin test (ASST), 28 CU patients with negative ASST (20 females and 8 males) as well as from healthy subjects matched by sex and age. The plasma level of uPA and suPAR antigens, PAI-1 activity did not differ significantly among the three subjects groups. The data obtained suggest that CU patients showing positive response to ASST have plasma profile of the urokinase system-associated proteins, which is not markedly different as compared with CU patients with negative ASST as well as healthy subjects. Our findings have also confirmed the earlier studies, suggesting that systemic fibrinolysis may not be involved in chronic urticaria.
Collapse
Affiliation(s)
- Alicja Kasperska-Zajac
- Clinical Department of Internal Diseases, Allergology and Clinical Immunology, Medical University of Silesia, ul. 3-go Maja 13-15, 41-800, Zabrze, Katowice, Poland.
| | | | | |
Collapse
|