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American Journal of Physiology. Cell... Jul 2022Corneal fibroblasts are embedded within an extracellular matrix composed largely of collagen type 1, proteoglycans, and other proteins in the corneal stroma, and their...
Corneal fibroblasts are embedded within an extracellular matrix composed largely of collagen type 1, proteoglycans, and other proteins in the corneal stroma, and their morphology and function are subject to continuous regulation by collagen. During wound healing and in various pathological conditions, corneal fibroblasts differentiate into myofibroblasts characterized by the expression of α-smooth muscle actin (α-SMA). Endo180, also known as urokinase-type plasminogen activator (uPA) receptor-associated protein (uPARAP), is a collagen receptor. Here we investigated whether targeting of Endo180 and the uPA receptor (uPAR) by uPA might play a role in the regulation of α-SMA expression by culturing corneal fibroblasts derived from uPA-deficient () or wild-type () mice in a collagen gel or on plastic. The expression of α-SMA was upregulated, the amounts of full-length Endo180 and uPAR were increased, and the levels of both transforming growth factor-β (TGF-β) expression and Smad3 phosphorylation were higher in corneal fibroblasts compared with cells under the collagen gel culture condition. Antibodies to Endo180 inhibited these effects of uPA deficiency on α-SMA and TGF-β expression, whereas a TGF-β signaling inhibitor blocked the effects on Smad3 phosphorylation and α-SMA expression. Our results suggest that uPA deficiency might promote the interaction between collagen and Endo180 and thereby increase α-SMA expression in a manner dependent on TGF-β signaling. Expression of α-SMA is thus negatively regulated by uPA through targeting of Endo180 and uPAR.
Topics: Actins; Animals; Collagen; Fibroblasts; Mice; Muscle, Smooth; Receptors, Mitogen; Transforming Growth Factor beta; Urokinase-Type Plasminogen Activator
PubMed: 35649252
DOI: 10.1152/ajpcell.00432.2021 -
Proceedings of the National Academy of... Jan 2023The urokinase-type plasminogen activator (uPA) system consists of the proteinase uPA, its receptor (PLAUR/uPAR). Under physiological conditions, uPA and PLAUR are...
The urokinase-type plasminogen activator (uPA) system consists of the proteinase uPA, its receptor (PLAUR/uPAR). Under physiological conditions, uPA and PLAUR are predominantly expressed by blood cells, including neutrophils, monocytes, and macrophages, and play important roles in cell activation, adhesion, migration, and extravasation. Here, we report that PLAUR, which is highly expressed in macrophages and dendritic cells (DCs) but hardly expressed in CD4 T cells, inhibits the release of HIV-1 progeny virions from the cell membrane. Silencing markedly enhanced the transmission of HIV-1 in macrophages and DCs. We further demonstrated that PLAUR is localized at the cell membrane to block the release of HIV-1 virions. Interestingly, we found that uPA compromises the PLAUR-mediated inhibition to slightly enhance HIV-1 production in primary macrophages and DCs. In the absence of PLAUR, this enhanced effect induced by uPA is abrogated. In conclusion, PLAUR is a new anti-HIV-1 protein produced in both macrophages and DCs where it inhibits HIV-1 transmission. This discovery may provide a novel therapeutic target for combating HIV.
Topics: Humans; Cell Membrane; HIV-1; Receptors, Urokinase Plasminogen Activator; Urokinase-Type Plasminogen Activator; Virion; Virus Replication
PubMed: 36638209
DOI: 10.1073/pnas.2212991120 -
World Neurosurgery Jun 2021The current treatment options for chronic subdural hematoma (CSDH) include burr hole drainage, twist drill drainage, and craniotomy with or without postoperative...
BACKGROUND
The current treatment options for chronic subdural hematoma (CSDH) include burr hole drainage, twist drill drainage, and craniotomy with or without postoperative catheter drainage. Although generally effective, these treatments have continued to be complicated by recurrence, especially in partially hemolyzed or septated hematomas. Recently, interest in the use of fibrinolytic agents as an adjunct to surgical treatment to address this limitation has been increasing. We conducted a systematic review, focusing on the efficacy and safety profile of fibrinolytic agents and compared the different fibrinolytic agents.
METHODS
The PubMed, EMBASE, CINAHL Plus, and Cochrane Library databases were searched for trials relevant to fibrinolytic administration in the treatment of CSDH. The findings are reported in accordance with the PRISMA (preferred reporting items for systematic reviews and meta-analyses) guidelines. The data from 1702 subjects from 6 retrospective observational studies were qualitatively analyzed. In addition, we included 11 case series and reports for discussion.
RESULTS
For 1449 patients, the use of urokinase or tissue plasminogen activator improved hematoma drainage and shortened the hospital stay (7.04 days), with an overall hematoma recurrence rate of 1.59%. The incidence of infection, seizure, and intracranial bleeding was 3.18%, 0.80%, and 0.41%, respectively, which compared favorably with previously reported findings for surgical drainage without the use of fibrinolytic agents.
CONCLUSIONS
The routine use of intrathecal urokinase and tissue plasminogen activator could be a new direction in the management of CSDH. Conclusive clinical evidence is lacking, however, and further prospective controlled studies are warranted to confirm the benefit and safety of this treatment strategy and to identify the optimal agent and dosing regimen.
Topics: Chemotherapy, Adjuvant; Craniotomy; Fibrinolytic Agents; Hematoma, Subdural, Chronic; Humans; Injections, Spinal; Tissue Plasminogen Activator; Urokinase-Type Plasminogen Activator
PubMed: 33722722
DOI: 10.1016/j.wneu.2021.03.029 -
No Shinkei Geka. Neurological Surgery Sep 2021Traumatic brain injury(TBI)is associated with coagulation and fibrinolytic disorder. It is characterized by consumptive coagulopathy and secondary hyperfibrinolysis...
Traumatic brain injury(TBI)is associated with coagulation and fibrinolytic disorder. It is characterized by consumptive coagulopathy and secondary hyperfibrinolysis associated with hypercoagulability and by hyperfibrinolysis due to the release of tissue plasminogen activator from the injured brain. Thrombin antithrombin III complex, a coagulation parameter, is abnormally high immediately after TBI and declines 6 hours after TBI. Fibrinogen, a coagulation factor, is rapidly consumed and degraded within 3 hours of TBI. D-dimer, a fibrinolytic parameter, is abnormally high on arrival at the hospital and reaches its maximum value 3 hours after TBI; during this time, bleeding tendency increases. Plasminogen activator inhibitor-1, a parameter of fibrinolysis shutdown, peaks at 6 hours after TBI. D-dimer is also known to be a prognostic factor. Patients with a high D-dimer level despite a good level of consciousness on admission are more likely to be "talk and deteriorate." Administration of tranexamic acid, an anti-fibrinolytic agent, early in the acute phase of TBI may reduce mortality. Fresh frozen plasma transfusion should be performed within 3 hours of TBI with monitoring of fibrinogen levels, and the administration dose should be set with a target fibrinogen level of ≧ 150 mg/dL. However, excessive administration should also be avoided. Thus, in the acute phase of TBI, coagulation and fibrinolytic activity changes dynamically and may adversely affect the complicated injury; therefore, monitoring coagulation and fibrinolytic parameters while conducting treatment is recommended.
Topics: Blood Component Transfusion; Humans; Plasma; Tissue Plasminogen Activator
PubMed: 34615754
DOI: 10.11477/mf.1436204477 -
Molecular Pharmaceutics Jun 2023The aberrant proteolytic landscape of the tumor microenvironment is a key contributor of cancer progression. Overexpression of urokinase plasminogen activator (uPA)... (Review)
Review
The aberrant proteolytic landscape of the tumor microenvironment is a key contributor of cancer progression. Overexpression of urokinase plasminogen activator (uPA) and/or its associated cell-surface receptor (uPAR) in tumor versus normal tissue is significantly associated with worse clinicopathological features and poorer patient survival across multiple cancer types. This is linked to mechanisms that facilitate tumor cell invasion and migration, via direct and downstream activation of various proteolytic processes that degrade the extracellular matrix─ultimately leading to metastasis. Targeting uPA has thus long been considered an attractive anticancer strategy. However, poor bioavailability of several uPA-selective small-molecule inhibitors has limited early clinical progress. Nanodelivery systems have emerged as an exciting method to enhance the pharmacokinetic (PK) profile of existing chemotherapeutics, allowing increased circulation time, improved bioavailability, and targeted delivery to tumor tissue. Combining uPA inhibitors with nanoparticle-based delivery systems thus offers a remarkable opportunity to overcome existing PK challenges associated with conventional uPA inhibitors, while leveraging potent candidates into novel targeted nanotherapeutics for an improved anticancer response in uPA positive tumors.
Topics: Humans; Neoplasm Invasiveness; Receptors, Urokinase Plasminogen Activator; Tumor Microenvironment; Urokinase-Type Plasminogen Activator
PubMed: 37119285
DOI: 10.1021/acs.molpharmaceut.3c00055 -
Scientific Reports Apr 2021Plasmin is the key enzyme in fibrinolysis. Upon interaction with plasminogen activators, the zymogen plasminogen is converted to active plasmin. Some studies indicate...
Plasmin is the key enzyme in fibrinolysis. Upon interaction with plasminogen activators, the zymogen plasminogen is converted to active plasmin. Some studies indicate plasminogen activation is regulated by cation-independent mannose 6-phosphate receptor (CI-MPR), a protein that facilitates lysosomal enzyme trafficking and insulin-like growth factor 2 downregulation. Plasminogen regulation may be accomplished by CI-MPR binding to plasminogen or urokinase plasminogen activator receptor. We asked whether other members of the plasminogen activation system, such as tissue plasminogen activator (tPA), also interact with CI-MPR. Because tPA is a glycoprotein with three N-linked glycosylation sites, we hypothesized that tPA contains mannose 6-phosphate (M6P) and binds CI-MPR in a M6P-dependent manner. Using surface plasmon resonance, we found that two sources of tPA bound the extracellular region of human and bovine CI-MPR with low-mid nanomolar affinities. Binding was partially inhibited with phosphatase treatment or M6P. Subsequent studies revealed that the five N-terminal domains of CI-MPR were sufficient for tPA binding, and this interaction was also partially mediated by M6P. The three glycosylation sites of tPA were analyzed by mass spectrometry, and glycoforms containing M6P and M6P-N-acetylglucosamine were identified at position N448 of tPA. In summary, we found that tPA contains M6P and is a CI-MPR ligand.
Topics: Acetylglucosamine; Animals; CHO Cells; Cells, Cultured; Cricetulus; Glycoproteins; Humans; Insulin-Like Growth Factor II; Ligands; Mannosephosphates; Phosphorylation; Protein Binding; Protein Interaction Domains and Motifs; Receptor, IGF Type 2; Sf9 Cells; Spodoptera; Tissue Plasminogen Activator
PubMed: 33859256
DOI: 10.1038/s41598-021-87579-z -
Journal of Visualized Experiments : JoVE Sep 2023A model of persisting lower back pain can be induced in mice with the simple methodology described herein. Step-by-step methods for simple, rapid induction of a...
A model of persisting lower back pain can be induced in mice with the simple methodology described herein. Step-by-step methods for simple, rapid induction of a persisting back pain model in mice are provided here using an injection of urokinase-type plasminogen activator (urokinase), a serine protease present in humans and other animals. The methodology for induction of persisting lower back pain in mice involves a simple injection of urokinase along the ligamentous insertion region of the lumbar spine. The urokinase inflammatory agent activates plasminogen to plasmin. Typically, the model can be induced within 10 min and hypersensitivity persists for at least 8 weeks. Hypersensitivity, gait disturbance, and other standard anxiety- and depression-like measures can be tested in the persisting model. Back pain is the most prevalent type of pain. To improve awareness of back pain, the International Association for the Study of Pain (IASP) named 2021 the "Global Year about Back Pain" and 2022 the "Global Year for Translating Pain Knowledge to Practice." One limitation of the therapeutic advancement of pain therapeutics is the lack of suitable models for testing persistent and chronic pain. The features of this model are suitable for testing potential therapeutics aimed at the reduction of back pain and its ancillary characteristics, contributing to IASP's naming 2022 as the Global Year for Translating Pain Knowledge to Practice.
Topics: Humans; Animals; Mice; Urokinase-Type Plasminogen Activator; Low Back Pain; Back Pain; Serine Proteases; Chronic Pain; Disease Models, Animal; Hypersensitivity
PubMed: 37677033
DOI: 10.3791/63997 -
F&S Science Feb 2023To study the role of interleukin (IL)-1β and the plasminogen activating (PA) system members in endometriotic stromal cell (ESC) migration/invasion.
OBJECTIVE
To study the role of interleukin (IL)-1β and the plasminogen activating (PA) system members in endometriotic stromal cell (ESC) migration/invasion.
DESIGN
Primary cultures of ESCs.
SETTING
Tertiary referral center for endometriosis and pelvic pain.
PATIENT(S)
Patients with surgically excised endometriosis.
INTERVENTION(S)
Interleukin-1β stimulation of primary cultures of ESCs and knockdown of the PA system members urokinase plasminogen activator (uPA), uPA receptor, and plasminogen activator inhibitor-1 (PAI-1).
MAIN OUTCOME MEASURE(S)
Invasion/migration assays.
RESULT(S)
In primary cultures, IL-1β-stimulated ESC production of the PA system members uPA, uPA receptor, and PAI-1. Interleukin-1β also enhanced ESC migration and invasion, and these effects were inhibited by the IL-1 receptor-1 antagonist anakinra. Knockdown of each of the 3 PA system members also inhibited ESC migration and invasion. Knockdown of these PA system members further attenuated the impact of IL-1β on migration and invasion, suggesting that they mediated the promigration and proinvasion effects of IL-1β. To supplement the cell culture work, immunohistochemistry was performed on tissue sections of endometriotic epithelium/stroma: uPA, PAI-1, and IL-1β histoscores were not found to be correlated with each other.
CONCLUSION(S)
In primary cultures of ESCs, IL-1β induces migration and invasion, which is mediated by PA system members and inhibited by the drug anakinra. However, the immunohistochemistry expression of IL-1β, urokinase plasminogen inhibitor-1, and PAI-1 were not correlated, suggesting other regulatory mechanisms for PA system members. Inhibition of IL-1β (e.g., with anakinra) may have potential as a novel treatment approach for the migration/invasion of endometriosis.
Topics: Female; Humans; Interleukin-1beta; Plasminogen Activator Inhibitor 1; Urokinase-Type Plasminogen Activator; Plasminogen; Endometriosis; Interleukin 1 Receptor Antagonist Protein; Cell Movement
PubMed: 36152991
DOI: 10.1016/j.xfss.2022.09.004 -
The European Journal of Neuroscience Apr 2020Epileptogenesis progressively leads to the rearrangement of normal neuronal networks into more excitable ones and can be viewed as a form of neuroplasticity, the...
Epileptogenesis progressively leads to the rearrangement of normal neuronal networks into more excitable ones and can be viewed as a form of neuroplasticity, the molecular mechanisms of which still remain obscure. Here, we studied pentylenetetrazole seizure-induced regulation of genes for plasminogen activator system in the mouse brain. We found that expression of tissue plasminogen activator (tPA) and urokinase receptor (uPAR) mRNA was strongly increased in the mouse cerebral cortex, hippocampus, striatum and amygdala as early as 3 hr after pentylenetetrazole seizures. Such early activity-induced expression of uPAR in the central nervous system has not been demonstrated before. uPAR mRNA accumulation was followed by elevation of uPAR protein, indicating a complete transcription-translation process. Both tPA gene induction and uPAR gene induction were independent of the protein synthesis, suggesting that they are regulated by neural activity as immediate-early genes. In contrast to tPA and uPAR genes, the expression of which returned to the basal level 6 hr following seizures, urokinase and plasminogen activator inhibitor-1 gene expression showed a delayed activation only at 3 days after seizures. In conclusion, our results suggest an important sensitivity of the brain plasminogen activator system to seizure activity which raises the question of its role in activity-dependent neural tissue remodeling in pathological and normal conditions.
Topics: Animals; Brain; Genes, Immediate-Early; Mice; Pentylenetetrazole; Receptors, Urokinase Plasminogen Activator; Seizures; Tissue Plasminogen Activator; Urokinase-Type Plasminogen Activator
PubMed: 31587391
DOI: 10.1111/ejn.14584 -
Journal of Pharmacy Practice Feb 2020To critically evaluate the published literature assessing the safety and efficacy of thrombolytic therapy for massive and submassive pulmonary embolism (PE). (Review)
Review
OBJECTIVE
To critically evaluate the published literature assessing the safety and efficacy of thrombolytic therapy for massive and submassive pulmonary embolism (PE).
METHODS
A search of human trials in the English-language (September 2017) was conducted through the MEDLINE database using the following terms: PE, tissue plasminogen activator, tenecteplase, and alteplase. 67 unique articles were identified, of which 24 clinical trials discussing clinical outcomes related to administration of either intravenous tenecteplase or alteplase were included.
RESULTS
Thrombolytic therapy with anticoagulation significantly reduced mortality compared to anticoagulation alone in massive PE. In submassive PE, thrombolytics reduced the rate of right ventricular dysfunction and hemodynamic collapse; however, there is an increased risk of major and minor bleeding with no benefit on long-term functional outcomes.
CONCLUSIONS
Patients with massive PE should receive thrombolytics when no major contraindications to therapy exist. Patients with submassive PE at highest risk for progression to hemodynamic instability should receive anticoagulation and be monitored for clinical deterioration. If an imminent risk of hemodynamic instability or cardiac arrest occurs, thrombolytics should be administered if no contraindications exist. Net mortality benefit and risk of bleeding must be considered when deciding to administer thrombolytic therapy in massive or submassive PE.
Topics: Anticoagulants; Drug Therapy, Combination; Fibrinolytic Agents; Hemorrhage; Humans; Pulmonary Embolism; Risk Assessment; Severity of Illness Index; Tenecteplase; Thrombolytic Therapy; Tissue Plasminogen Activator; Treatment Outcome
PubMed: 29673293
DOI: 10.1177/0897190018767769