Pulmonary delivery of the broad-spectrum matrix metalloproteinase inhibitor marimastat diminishes multiwalled carbon nanotube-induced circulating bioactivity without reducing pulmonary inflammation

Abstract Background Multiwalled carbon nanotubes (MWCNT) are an increasingly utilized engineered nanomaterial that pose the potential for significant risk of exposure-related health outcomes. The mechanism(s) underlying MWCNT-induced toxicity to extrapulmonary sites are still being defined. MWCNT-induced serum-borne bioactivity appears to dysregulate systemic endothelial cell function. The serum compositional changes after MWCNT exposure have been identified as a surge of fragmented endogenous peptides, likely derived from matrix metalloproteinase (MMP) activity. In the present study, we utilize a broad-spectrum MMP inhibitor, Marimastat, along with a previously described oropharyngeal aspiration model of MWCNT administration to investigate the role of MMPs in MWCNT-derived serum peptide generation and endothelial bioactivity. Results C57BL/6 mice were treated with Marimastat or vehicle by oropharyngeal aspiration 1 h prior to MWCNT treatment. Pulmonary neutrophil infiltration and total bronchoalveolar lavage fluid protein increased independent of MMP blockade. The lung cytokine profile similarly increased following MWCNT exposure for major inflammatory markers (IL-1β, IL-6, and TNF-α), with minimal impact from MMP inhibition. However, serum peptidomic analysis revealed differential peptide compositional profiles, with MMP blockade abrogating MWCNT-derived serum peptide fragments. The serum, in turn, exhibited differential potency in terms of inflammatory bioactivity when incubated with primary murine cerebrovascular endothelial cells. Serum from MWCNT-treated mice led to inflammatory responses in endothelial cells that were significantly blunted with serum from Marimastat-treated mice. Conclusions Thus, MWCNT exposure induced pulmonary inflammation that was largely independent of MMP activity but generated circulating bioactive peptides through predominantly MMP-dependent pathways. This MWCNT-induced lung-derived bioactivity caused pathological consequences of endothelial inflammation and barrier disruption.

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Role of Matrix Metalloproteinases in Multiwalled Carbon Nanotube-mediated Pulmonary and Systemic Inflammatory Activation

10.21203/rs.3.rs-406038/v1 ◽

2021 ◽

Author(s):

Tamara L. Young ◽

Ekaterina Mostovenko ◽

Jesse L. Denson ◽

Jessica G. Begay ◽

Selita N. Lucas ◽

...

Keyword(s):

Endothelial Cells ◽

Cell Function ◽

Inflammatory Responses ◽

Significant Risk ◽

Dependent Manner ◽

Multiwalled Carbon ◽

Endothelial Cell Function ◽

Serum Peptide ◽

Oropharyngeal Aspiration

Abstract Background: Multiwalled carbon nanotubes (MWCNT) are an increasingly utilized engineered nanomaterial that pose the potential for significant risk of exposure-related health outcomes. The mechanism(s) underlying MWCNT-induced toxicity to extrapulmonary sites are still being defined. MWCNT-induced serum-borne bioactivity appears to dysregulate systemic endothelial cell function. The serum compositional changes after MWCNT exposure have been identified as a surge of fragmented endogenous peptides, likely derived from matrix metalloproteinase (MMP) activity. In the present study, we utilize a broad-spectrum MMP inhibitor, Marimastat, along with a previously described oropharyngeal aspiration model of MWCNT administration to investigate the role of MMPs in MWCNT-derived serum peptide generation and endothelial bioactivity. Results: C57BL/6 mice were treated with Marimastat or vehicle by oropharyngeal aspiration 1h prior to MWCNT treatment. Pulmonary neutrophil infiltration and total bronchoalveolar lavage fluid protein increased in a dose-dependent manner, independent of MMP blockade. The lung cytokine profile similarly increased following MWCNT exposure for major inflammatory markers (IL-1β, IL-6, and TNFα), with minimal impact from MMP inhibition. However, serum peptidomic analysis revealed differential peptide compositional profiles, with MMP blockade abrogating MWCNT-derived serum peptide fragments. The serum, in turn, exhibited differential potency in terms of inflammatory bioactivity when incubated with primary murine cerebrovascular endothelial cells. Serum from MWCNT-treated mice led to inflammatory responses in endothelial cells that were significantly blunted with serum from Marimastat-treated mice. Conclusion: Thus, MWCNT exposure induced pulmonary inflammation that was largely independent of MMP activity, but generated circulating bioactive peptides through predominantly MMP-dependent pathways. This MWCNT-induced lung-derived bioactivity caused pathological consequences of endothelial inflammation and barrier disruption.

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Loss of endothelial sulfatase-1 after experimental sepsis attenuates subsequent pulmonary inflammatory responses

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00175.2019 ◽

2019 ◽

Vol 317 (5) ◽

pp. L667-L677 ◽

Cited By ~ 2

Author(s):

Kaori Oshima ◽

Xiaorui Han ◽

Yilan Ouyang ◽

Rana El Masri ◽

Yimu Yang ◽

...

Keyword(s):

Endothelial Cells ◽

Heparan Sulfate ◽

Inflammatory Responses ◽

Pulmonary Inflammation ◽

Endothelial Glycocalyx ◽

Experimental Sepsis ◽

Pulmonary Response ◽

Increased Risk ◽

Heparan Sulfates ◽

Hospital Acquired

Sepsis patients are at increased risk for hospital-acquired pulmonary infections, potentially due to postseptic immunosuppression known as the compensatory anti-inflammatory response syndrome (CARS). CARS has been attributed to leukocyte dysfunction, with an unclear role for endothelial cells. The pulmonary circulation is lined by an endothelial glycocalyx, a heparan sulfate-rich layer essential to pulmonary homeostasis. Heparan sulfate degradation occurs early in sepsis, leading to lung injury. Endothelial synthesis of new heparan sulfates subsequently allows for glycocalyx reconstitution and endothelial recovery. We hypothesized that remodeling of the reconstituted endothelial glycocalyx, mediated by alterations in the endothelial machinery responsible for heparan sulfate synthesis, contributes to CARS. Seventy-two hours after experimental sepsis, coincident with glycocalyx reconstitution, mice demonstrated impaired neutrophil and protein influx in response to intratracheal lipopolysaccharide (LPS). The postseptic reconstituted glycocalyx was structurally remodeled, with enrichment of heparan sulfate disaccharides sulfated at the 6- O position of glucosamine. Increased 6- O-sulfation coincided with loss of endothelial sulfatase-1 (Sulf-1), an enzyme that specifically removes 6- O-sulfates from heparan sulfate. Intravenous administration of Sulf-1 to postseptic mice restored the pulmonary response to LPS, suggesting that loss of Sulf-1 was necessary for postseptic suppression of pulmonary inflammation. Endothelial-specific knockout mice demonstrated that loss of Sulf-1 was not sufficient to induce immunosuppression in non-septic mice. Knockdown of Sulf-1 in human pulmonary microvascular endothelial cells resulted in downregulation of the adhesion molecule ICAM-1. Taken together, our study indicates that loss of endothelial Sulf-1 is necessary for postseptic suppression of pulmonary inflammation, representing a novel endothelial contributor to CARS.

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Regulation of Matrix Metalloproteinase in Vascularization of Endothelial Cells On Extracellular Matrix Secreted By Mesenchymal Stem Cell Sheet

10.21203/rs.3.rs-167880/v1 ◽

2021 ◽

Author(s):

Lina Ren ◽

Jinda Li ◽

Dan Yang ◽

Xuan Zhang ◽

Yuan Shen ◽

...

Keyword(s):

Stem Cells ◽

Endothelial Cells ◽

Stem Cell ◽

Mesenchymal Stem Cell ◽

Matrix Metalloproteinase ◽

Broad Spectrum ◽

Cell Sheet ◽

Immunofluorescence Staining ◽

Rt Pcr ◽

Mesenchymal Stem Cell Sheet

Abstract Background: We have previously developed a prevascularized platform by seeding endothelial cells on the mesenchymal stem cell sheet and the technology provides a promising vascularization potential for wide applications in tissue engineering. However, the mechanism behind the interactions between stem cell sheet-derived matrix and endothelial cells (ECs) are still unknown. The purpose of this study is to investigate the role of MMPs in angiogenesis process in vitro.Methods: Human bone marrow-derived mesenchymal stem cells (hBMSCs) were cultured continually to form a thin cell sheet, and then green fluorescent protein (GFP)-labeled human umbilical vein endothelial cells (HUVECs) were seeded on the cell sheet to form a prevascularized system. Fibronectin, collagen I and MMPs immunofluorescence staining were used to observe their interaction changes. After that, HUVECs and hBMSCs were sorted by flow cytometry and RT-PCR was performed to quantify related gene expression of ECs and stem cells. In order to further explore the possible mechanism, the broad-spectrum MMPs inhibitor GM6001 was added into the medium to observe the effects of vascular network formation. Results: The immunofluorescence staining results showed that HUVECs secreted MMP-2 and membrane-type matrix metalloproteinase-1 (MT1-MMP) in the early stage of angiogenesis. MMPs degraded and remodeled the ECM to create space for vascular formation. HUVECs also promoted hBMSCs to secrete tissue inhibitors of metalloproteinase-2 (TIMP-2), which can stabilize blood vessels by avoiding excessive vascularization or vascular degradation in the later stage. Thus, HUVECs/hBMSCs co-culture group were beneficial for angiogenesis than the other two control groups. With the addition of the broad-spectrum matrix metalloproteinase inhibitor GM6001, RT-PCR results showed that the formation of EC lumen was blocked, while the mean fluorescence density of fibronectin and collagen I were increased.Conclusions: our study suggests that the MMP-2, MT1-MMP and TIMP-2 can synergistically regulate the angiogenesis. In addition, HUVECs and hBMSCs interact with each other in the process of vascular formation, and jointly regulate and promote the construction of lumen. These results also indicate a promising potential using the hBMSCs sheet as a platform for broad applications in engineering vascularized tissues.

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CD40/CD40L contributes to hypercholesterolemia-induced microvascular inflammation

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00962.2008 ◽

2009 ◽

Vol 296 (3) ◽

pp. H689-H697 ◽

Cited By ~ 30

Author(s):

Karen Y. Stokes ◽

LeShanna Calahan ◽

Candiss M. Hamric ◽

Janice M. Russell ◽

D. Neil Granger

Keyword(s):

Oxidative Stress ◽

T Cells ◽

T Cell ◽

Blood Cell ◽

Cell Function ◽

Inflammatory Responses ◽

Microvascular Dysfunction ◽

Cd40 Ligand ◽

Scid Mice ◽

Endothelial Cell Function

Hypercholesterolemia is associated with phenotypic changes in endothelial cell function that lead to a proinflammatory and prothrombogenic state in different segments of the microvasculature. CD40 ligand (CD40L) and its receptor CD40 are ubiquitously expressed and mediate inflammatory responses and platelet activation. The objective of this study was to determine whether CD40/CD40L, in particular T-cell CD40L, contributes to microvascular dysfunction induced by hypercholesterolemia. Intravital microscopy was used to quantify blood cell adhesion in cremasteric postcapillary venules, endothelium-dependent vasodilation responses in arterioles, and microvascular oxidative stress in wild-type (WT) C57BL/6, CD40-deficient (−/−), CD40L−/−, or severe combined immune deficient (SCID) mice placed on a normal (ND) or high-cholesterol (HC) diet for 2 wk. WT-HC mice exhibited an exaggerated leukocyte and platelet recruitment in venules and impaired vasodilation responses in arterioles compared with ND counterparts. A deficiency of CD40, CD40L, or lymphocytes attenuated these responses to HC. The HC phenotype was rescued in CD40L−/− and SCID mice by a transfer of WT T cells. Bone marrow chimeras revealed roles for both vascular- and blood cell-derived CD40 and CD40L in the HC-induced vascular responses. Hypercholesterolemia induced an oxidative stress in both arterioles and venules of WT mice, which was abrogated by either CD40 or CD40L deficiency. The transfer of WT T cells into CD40L−/− mice restored the oxidative stress. These results implicate CD40/CD40L interactions between circulating cells and the vascular wall in both the arteriolar and venular dysfunction elicited by hypercholesterolemia and identify T-cell-associated CD40L as a key mediator of these responses.

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Investigation of Wall Shear Stress in Cardiovascular Research and in Clinical Practice—From Bench to Bedside

International Journal of Molecular Sciences ◽

10.3390/ijms22115635 ◽

2021 ◽

Vol 22 (11) ◽

pp. 5635

Author(s):

Katharina Urschel ◽

Miyuki Tauchi ◽

Stephan Achenbach ◽

Barbara Dietel

Keyword(s):

Endothelial Cells ◽

Shear Stress ◽

Wall Shear Stress ◽

Cell Function ◽

Computational Techniques ◽

Apical Surface ◽

Cardiovascular Research ◽

Endothelial Cell Function ◽

Wall Shear

In the 1900s, researchers established animal models experimentally to induce atherosclerosis by feeding them with a cholesterol-rich diet. It is now accepted that high circulating cholesterol is one of the main causes of atherosclerosis; however, plaque localization cannot be explained solely by hyperlipidemia. A tremendous amount of studies has demonstrated that hemodynamic forces modify endothelial athero-susceptibility phenotypes. Endothelial cells possess mechanosensors on the apical surface to detect a blood stream-induced force on the vessel wall, known as “wall shear stress (WSS)”, and induce cellular and molecular responses. Investigations to elucidate the mechanisms of this process are on-going: on the one hand, hemodynamics in complex vessel systems have been described in detail, owing to the recent progress in imaging and computational techniques. On the other hand, investigations using unique in vitro chamber systems with various flow applications have enhanced the understanding of WSS-induced changes in endothelial cell function and the involvement of the glycocalyx, the apical surface layer of endothelial cells, in this process. In the clinical setting, attempts have been made to measure WSS and/or glycocalyx degradation non-invasively, for the purpose of their diagnostic utilization. An increasing body of evidence shows that WSS, as well as serum glycocalyx components, can serve as a predicting factor for atherosclerosis development and, most importantly, for the rupture of plaques in patients with high risk of coronary heart disease.

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Reversal of the Inflammatory Responses in Fabry Patient iPSC-Derived Cardiovascular Endothelial Cells by CRISPR/Cas9-Corrected Mutation

International Journal of Molecular Sciences ◽

10.3390/ijms22052381 ◽

2021 ◽

Vol 22 (5) ◽

pp. 2381

Author(s):

Hui-Yung Song ◽

Yi-Ping Yang ◽

Yueh Chien ◽

Wei-Yi Lai ◽

Yi-Ying Lin ◽

...

Keyword(s):

Endothelial Cells ◽

Inflammatory Responses ◽

Late Onset ◽

Critical Role ◽

Vascular Complications ◽

Digital Pcr ◽

Mapk Signaling ◽

Autophagic Flux ◽

Fabry Patient ◽

Genomic Editing

The late-onset type of Fabry disease (FD) with GLA IVS4 + 919G > A mutation has been shown to lead to cardiovascular dysfunctions. In order to eliminate variations in other aspects of the genetic background, we established the isogenic control of induced pluripotent stem cells (iPSCs) for the identification of the pathogenetic factors for FD phenotypes through CRISPR/Cas9 genomic editing. We adopted droplet digital PCR (ddPCR) to efficiently capture mutational events, thus enabling isolation of the corrected FD from FD-iPSCs. Both of these exhibited the characteristics of pluripotency and phenotypic plasticity, and they can be differentiated into endothelial cells (ECs). We demonstrated the phenotypic abnormalities in FD iPSC-derived ECs (FD-ECs), including intracellular Gb3 accumulation, autophagic flux impairment, and reactive oxygen species (ROS) production, and these abnormalities were rescued in isogenic control iPSC-derived ECs (corrected FD-ECs). Microarray profiling revealed that corrected FD-derived endothelial cells reversed the enrichment of genes in the pro-inflammatory pathway and validated the downregulation of NF-κB and the MAPK signaling pathway. Our findings highlighted the critical role of ECs in FD-associated vascular dysfunctions by establishing a reliable isogenic control and providing information on potential cellular targets to reduce the morbidity and mortality of FD patients with vascular complications.

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Type-2 Diabetes as a Risk Factor for Severe COVID-19 Infection

Microorganisms ◽

10.3390/microorganisms9061211 ◽

2021 ◽

Vol 9 (6) ◽

pp. 1211

Author(s):

Mahnaz Norouzi ◽

Shaghayegh Norouzi ◽

Alistaire Ruggiero ◽

Mohammad S. Khan ◽

Stephen Myers ◽

...

Keyword(s):

Type 2 Diabetes ◽

Risk Factor ◽

Inflammatory Responses ◽

Significant Risk ◽

Significant Risk Factor ◽

Diabetic Patients ◽

Angiotensin Converting Enzyme 2 ◽

Immune System Dysfunction ◽

Current Outbreak

The current outbreak caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), termed coronavirus disease 2019 (COVID-19), has generated a notable challenge for diabetic patients. Overall, people with diabetes have a higher risk of developing different infectious diseases and demonstrate increased mortality. Type 2 diabetes mellitus (T2DM) is a significant risk factor for COVID-19 progression and its severity, poor prognosis, and increased mortality. How diabetes contributes to COVID-19 severity is unclear; however, it may be correlated with the effects of hyperglycemia on systemic inflammatory responses and immune system dysfunction. Using the envelope spike glycoprotein SARS-CoV-2, COVID-19 binds to angiotensin-converting enzyme 2 (ACE2) receptors, a key protein expressed in metabolic organs and tissues such as pancreatic islets. Therefore, it has been suggested that diabetic patients are more susceptible to severe SARS-CoV-2 infections, as glucose metabolism impairments complicate the pathophysiology of COVID-19 disease in these patients. In this review, we provide insight into the COVID-19 disease complications relevant to diabetes and try to focus on the present data and growing concepts surrounding SARS-CoV-2 infections in T2DM patients.

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The ATP Receptors P2X7 and P2X4 Modulate High Glucose and Palmitate-Induced Inflammatory Responses in Endothelial Cells

PLoS ONE ◽

10.1371/journal.pone.0125111 ◽

2015 ◽

Vol 10 (5) ◽

pp. e0125111 ◽

Cited By ~ 24

Author(s):

Ramasri Sathanoori ◽

Karl Swärd ◽

Björn Olde ◽

David Erlinge

Keyword(s):

Endothelial Cells ◽

High Glucose ◽

Inflammatory Responses ◽

Atp Receptors

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Regulatory T Cells Protect Fine Particulate Matter-Induced Inflammatory Responses in Human Umbilical Vein Endothelial Cells

Mediators of Inflammation ◽

10.1155/2014/869148 ◽

2014 ◽

Vol 2014 ◽

pp. 1-14 ◽

Cited By ~ 6

Author(s):

Wen-cai Zhang ◽

Yan-ge Wang ◽

Zheng-feng Zhu ◽

Fang-qin Wu ◽

Yu-dong Peng ◽

...

Keyword(s):

T Cells ◽

Endothelial Cells ◽

Particulate Matter ◽

Inflammatory Cytokines ◽

Fine Particles ◽

Inflammatory Responses ◽

Umbilical Vein ◽

Fine Particulate Matter ◽

Fine Particulate ◽

Umbilical Vein Endothelial Cells

Objective. To investigate the role of CD4+CD25+T cells (Tregs) in protecting fine particulate matter (PM-) induced inflammatory responses, and its potential mechanisms.Methods. Human umbilical vein endothelial cells (HUVECs) were treated with graded concentrations (2, 5, 10, 20, and 40 µg/cm2) of suspension of fine particles for 24h. For coculture experiment, HUVECs were incubated alone, with CD4+CD25−T cells (Teff), or with Tregs in the presence of anti-CD3 monoclonal antibodies for 48 hours, and then were stimulated with or without suspension of fine particles for 24 hours. The expression of adhesion molecules and inflammatory cytokines was examined.Results. Adhesion molecules, including vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1), and inflammatory cytokines, such as interleukin (IL-) 6 and IL-8, were increased in a concentration-dependent manner. Moreover, the adhesion of human acute monocytic leukemia cells (THP-1) to endothelial cells was increased and NF-κB activity was upregulated in HUVECs after treatment with fine particles. However, after Tregs treatment, fine particles-induced inflammatory responses and NF-κB activation were significantly alleviated. Transwell experiments showed that Treg-mediated suppression of HUVECs inflammatory responses impaired by fine particles required cell contact and soluble factors.Conclusions. Tregs could attenuate fine particles-induced inflammatory responses and NF-κB activation in HUVECs.

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