Analysis of the Heavy-Chain Modification and TSG-6 Activity in Pathological Hyaluronan Matrices

Lauer, Mark E.; Loftis, Jacqueline; Motte, Carol A. de la; Hascall, Vincent C.

doi:10.1007/978-1-4939-1714-3_42

Cited by 24 publications

(22 citation statements)

References 4 publications

(4 reference statements)

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“…Hyaluronidase Extraction of Heavy Chains from Asthmatic Lung Tissue and BAL Fluid-This method has been previously described and is presented here with minor changes (25). Human lung tissue from a patient who died from acute severe asthma was cut with a scalpel and transferred to a preweighed 1.5-ml tube that was prechilled on dry ice.…”

Section: Methodsmentioning

confidence: 99%

Hyaluronan and Its Heavy Chain Modification in Asthma Severity and Experimental Asthma Exacerbation

Lauer

Majors

Comhair

et al. 2015

Journal of Biological Chemistry

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Background: Hyaluronan has been linked to asthma severity and inflammation. Results: We characterized the hyaluronan levels and its heavy chain modification in an experimental model of human asthma exacerbation. Conclusion: These data implicate hyaluronan and its heavy chain modification in human asthma severity. Significance: Repetitive asthma exacerbations exacerbate hyaluronan pathobiology, which contribute to the chronic inflammation associated with this disease.

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Section: Methodsmentioning

confidence: 99%

Hyaluronan and Its Heavy Chain Modification in Asthma Severity and Experimental Asthma Exacerbation

Lauer

Majors

Comhair

et al. 2015

Journal of Biological Chemistry

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“…This method was previously described (6,19). Human CF and control lung/bronchus tissue were processed and blotted using Bio-Rad nitrocellulose and Trans-Blot Turbo System (Biorad, Hercules, CA), probed with a rabbit polyclonal antibody against IaI (1:8,000 dilution), and the secondary was anti-rabbit 800CW (1:15:000; 926-32211; Li-Cor, Lincoln, NE).…”

Section: Ha-hc Analysis By Western Blotmentioning

confidence: 99%

Pathological Hyaluronan Matrices in Cystic Fibrosis Airways and Secretions

Matuska¹,

Comhair

Farver³

et al. 2016

Am J Respir Cell Mol Biol

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Hyaluronan (HA) has been used in treatment of cystic fibrosis (CF) via a nebulizer and has demonstrated success in clinical outcomes. HA is an important glycosaminoglycan that is cross-linked by heavy chains (HCs) from inter-α-inhibitor during inflammation. HC cross-linked HA (HC-HA) becomes significantly more adhesive for leukocytes than non-cross-linked HA, which can enhance inflammation. Our studies tested the hypothesis that HC-HA is present in CF airways and that altered ratios of HC-HA to its degradation into relatively lower molecular weight HA contribute to the pathophysiology of chronic inflammation in CF. We evaluated the distribution, levels, and size of HC-HA within CF, healthy, and diseased control lung, bronchus, and sputum tissues by histological and biochemical approaches. HC-HA was significantly elevated in CF, with deposits around the pulmonary vasculature, airway submucosa, and in the stroma of the submucosal glands. The increased infiltration of leukocyte populations correlated with the distribution of HC-HA matrices in the airways. Elevated lung tissue HC-HA correlated with decreased HA levels in CF mucus and sputum compared with controls, suggesting that aberrant degradation and cross-linking of HA in lung tissue is a unique feature of CF. The accumulation and degradation of proinflammatory HC-HA in CF lung tissue suggests that aberrant HA catabolism and cross-linking may contribute to chronic inflammation in airway tissues and affect mucus viscosity in CF airways.

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“…Detection of HA heavy chain (HC) modification. We utilized the protocol developed by Mark Lauer et al 33 . Frozen lung samples (50 mg) were homogenized in PBS for 90 seconds using 1.6 mm ceramic beads (OMNI, Kennesaw, GA).…”

mentioning

confidence: 99%

“…Membranes were then washed and developed with peroxidase enhanced chemiluminescence (ECL) reagent (Clarity ECL, BioRad) for 1 minute and imaged on a ChemiDoc MP. HC-HA complexes are impermeable to the gel, and thus HC was identified as a band alternately present (hyaluronidase-digested) and absent (undigested) at 74kD according to previously published protocols validating the antibody in mouse lungs 33 . Densitometry was performed on ImageJ and normalized to total protein.…”

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confidence: 99%

Extracellular Superoxide Dismutase Regulates Early Vascular Hyaluronan Remodeling in Hypoxic Pulmonary Hypertension

Tseng

Allawzi

et al. 2020

Sci Rep

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chronic hypoxia leads to pathologic remodeling of the pulmonary vasculature and pulmonary hypertension (PH). The antioxidant enzyme extracellular superoxide dismutase (SOD3) protects against hypoxia-induced pH. Hyaluronan (HA), a ubiquitous glycosaminoglycan of the lung extracellular matrix, is rapidly recycled at sites of vessel injury and repair. We investigated the hypothesis that SOD3 preserves HA homeostasis by inhibiting oxidative and enzymatic hyaluronidase-mediated HA breakdown. In SOD3-deficient mice, hypoxia increased lung hyaluronidase expression and activity, hyaluronan fragmentation, and effacement of HA from the vessel wall of small pulmonary arteries. Hyaluronan fragmentation corresponded to hypoxic induction of the cell surface hyaluronidase-2 (Hyal2), which was localized in the vascular media. Human pulmonary artery smooth muscle cells (HPASMCs) demonstrated hypoxic induction of Hyal2 and SOD-suppressible hyaluronidase activity, congruent to our observations in vivo. fragmentation of homeostatic high molecular weight HA promoted HpASMc proliferation in vitro, whereas pharmacologic inhibition of hyaluronidase activity prevented hypoxia-and oxidant-induced proliferation. Hypoxia initiates SOD3-dependent alterations in the structure and regulation of hyaluronan in the pulmonary vascular extracellular matrix. these changes occurred soon after hypoxia exposure, prior to appearance of pH, and may contribute to the early pathogenesis of this disease.Pulmonary hypertension (PH) is an incurable and fatal disease characterized by intimal thickening, muscular hypertrophy, excessive matrix deposition, and stiffening of arteries in the lung. Chronic hypoxia is a major cause and consequence of PH in humans. It is the most prevalent etiology of precapillary PH in humans (WHO Group 3 disease). The onset of PH in patients with hypoxia due to intrinsic lung disease portends a grim prognosis for survival, post-transplant outcome, and quality of life 1,2 . Therefore, there is an urgent need for research on the molecular pathogenesis of hypoxic pulmonary hypertension.One facet of this disease is the presence of high oxidative stress within the vessel wall and perivascular matrix 3 . Extracellular superoxide dismutase (SOD3) is crucial for neutralization of this stress. Global knockout 4 , selective deletion from smooth muscle cells 5 , and disrupted matrix-binding single nucleotide polymorphisms 6 of SOD3 result in a great susceptibility to hypoxia-induced vascular inflammation, fibrosis, and PH. On the other hand, lung-specific overexpression of SOD3 7 , adenoviral gene transfer of human SOD3 8 , and treatment with an inorganic metalloporphyrin SOD3 mimetic 9 are protective against PH in rodent models. In the advanced stages of PH in humans, SOD3 levels are epigenetically reduced by histone deacetylation 10 .There is a critical gap in our understanding of how the extracellular oxidative environment can lead to irreversible pulmonary vascular remodeling in chronic hypoxia. Amassing evidence points to a central role f...

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Analysis of the Heavy-Chain Modification and TSG-6 Activity in Pathological Hyaluronan Matrices

Cited by 24 publications

References 4 publications

Hyaluronan and Its Heavy Chain Modification in Asthma Severity and Experimental Asthma Exacerbation

Hyaluronan and Its Heavy Chain Modification in Asthma Severity and Experimental Asthma Exacerbation

Pathological Hyaluronan Matrices in Cystic Fibrosis Airways and Secretions

Extracellular Superoxide Dismutase Regulates Early Vascular Hyaluronan Remodeling in Hypoxic Pulmonary Hypertension

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