Tetrathiomolybdate Treatment Attenuates Bleomycin-Induced Angiogenesis and Lung Pathology in a Sheep Model of Pulmonary Fibrosis

Idiopathic pulmonary fibrosis (IPF) is a progressive chronic lung disease characterized by excessive extracellular matrix (ECM) deposition in the parenchyma of the lung. Accompanying the fibrotic remodeling, dysregulated angiogenesis has been observed and implicated in the development and progression of pulmonary fibrosis. Copper is known to be required for key processes involved in fibrosis and angiogenesis. We therefore hypothesized that lowering bioavailable serum copper with tetrathiomolybdate could be of therapeutic value for treating pulmonary fibrosis. This study aimed to investigate the effect of tetrathiomolybdate on angiogenesis and fibrosis induced in sheep lung segments infused with bleomycin. Twenty sheep received two fortnightly infusions of either bleomycin (3U), or saline (control) into two spatially separate lung segments. A week after the final bleomycin/saline infusions, sheep were randomly assigned into two groups (n = 10 per group) and received twice-weekly intravenous administrations of either 50 mg tetrathiomolybdate, or sterile saline (vehicle control), for 6 weeks. Vascular density, expressed as the percentage of capillary area to the total area of parenchyma, was determined in lung tissue sections immuno-stained with antibodies against CD34 and collagen type IV. The degree of fibrosis was assessed by histopathology scoring of H&E stained sections and collagen content using Masson’s trichrome staining. Lung compliance was measured via a wedged bronchoscope procedure prior to and 7 weeks following final bleomycin infusion. In this large animal model, we show that copper lowering by tetrathiomolybdate chelation attenuates both bleomycin-induced angiogenesis and pulmonary fibrosis. Moreover, tetrathiomolybdate treatment downregulates vascular endothelial growth factor (VEGF) expression, and improved lung function in bleomycin-induced pulmonary fibrosis. Tetrathiomolybdate also suppressed the accumulation of inflammatory cells in bronchoalveolar lavage fluid 2 weeks after bleomycin injury. The molecular mechanism(s) underpinning copper modulation of fibrotic pathways is an important area for future investigation, and it represents a potential therapeutic target for pulmonary fibrosis.

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The efficacy and safety of pinocembrin in a sheep model of bleomycin-induced pulmonary fibrosis

PLoS ONE ◽

10.1371/journal.pone.0260719 ◽

2021 ◽

Vol 16 (12) ◽

pp. e0260719

Author(s):

Habtamu B. Derseh ◽

Jason Q. D. Goodger ◽

Jean-Pierre Y. Scheerlinck ◽

Chrishan S. Samuel ◽

Ian E. Woodrow ◽

...

Keyword(s):

Animal Model ◽

Pulmonary Fibrosis ◽

Inflammatory Cells ◽

Lung Parenchyma ◽

Lung Compliance ◽

Large Animal Model ◽

Large Animal ◽

Lung Pathology ◽

Sheep Model ◽

Anti Inflammatory

The primary flavonoid, pinocembrin, is thought to have a variety of medical uses which relate to its reported anti-oxidant, anti-inflammatory, anti-microbial and anti-cancer properties. Some studies have reported that this flavonoid has anti-fibrotic activities. In this study, we investigated whether pinocembrin would impede fibrosis, dampen inflammation and improve lung function in a large animal model of pulmonary fibrosis. Fibrosis was induced in two localized lung segments in each of the 10 sheep participating in the study. This was achieved via two infusions of bleomycin delivered bronchoscopically at a two-week interval. Another lung segment in the same sheep was left untreated, and was used as a healthy control. The animals were kept for a little over 5 weeks after the final infusion of bleomycin. Pinocembrin, isolated from Eucalyptus leaves, was administered to one of the two bleomycin damaged lung segments at a dose of 7 mg. This dose was given once-weekly over 4-weeks, starting one week after the final bleomycin infusion. Lung compliance (as a measure of stiffness) was significantly improved after four weekly administrations of pinocembrin to bleomycin-damaged lung segments. There were significantly lower numbers of neutrophils and inflammatory cells in the bronchoalveolar lavage of bleomycin-infused lung segments that were treated with pinocembrin. Compared to bleomycin damaged lung segments without drug treatment, pinocembrin administration was associated with significantly lower numbers of immuno-positive CD8+ and CD4+ T cells in the lung parenchyma. Histopathology scoring data showed that pinocembrin treatment was associated with significant improvement in inflammation and overall pathology scores. Hydroxy proline analysis showed that the administration of pinocembrin did not reduce the increased collagen content that was induced by bleomycin in this model. Analyses of Masson’s Trichrome stained sections showed that pinocembrin treatment significantly reduced the connective tissue content in lung segments exposed to bleomycin when compared to bleomycin-infused lungs that did not receive pinocembrin. The striking anti-inflammatory and modest anti-fibrotic remodelling effects of pinocembrin administration were likely linked to the compound’s ability to improve lung pathology and functional compliance in this animal model of pulmonary fibrosis.

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Tetrathiomolybdate Treatment Attenuates Microvascular Remodeling in a Large Animal Model of Bleomycin Induced Pulmonary Fibrosis

10.1164/ajrccm-conference.2020.201.1_meetingabstracts.a2390 ◽

2020 ◽

Author(s):

H.B. Derseh ◽

K.U.E.U.E. Perera ◽

V.D.S.D.S. Nimanthi ◽

E. Koumoundouros ◽

C.N. Pagel ◽

...

Keyword(s):

Animal Model ◽

Pulmonary Fibrosis ◽

Large Animal Model ◽

Large Animal ◽

Microvascular Remodeling

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Abstract 13: Adenosine Receptor Activation During Extracorporeal Cardiopulmonary Resuscitation Improves Survival After Cardiac Arrest in Swine

Circulation ◽

10.1161/circ.140.suppl_2.13 ◽

2019 ◽

Vol 140 (Suppl_2) ◽

Author(s):

William Z Chancellor ◽

Dustin Money ◽

Jared P Beller ◽

Matthew R Byler ◽

Aimee Zhang ◽

...

Keyword(s):

Cardiac Arrest ◽

Cardiopulmonary Resuscitation ◽

Troponin I ◽

Circulatory Arrest ◽

Receptor Activation ◽

Large Animal Model ◽

Saline Control ◽

Large Animal ◽

Extracorporeal Cardiopulmonary Resuscitation ◽

A2ar Agonist

Introduction: Despite advances in resuscitation protocols, including the addition of Extracorporeal Cardiopulmonary Resuscitation (ECPR), survival after cardiac arrest remains less than 40% and novel methods are needed to attenuate global injury and improve outcomes. ATL1223, an adenosine 2A receptor (A2AR) agonist has been shown to attenuate organ specific reperfusion injury by modulating the interaction of inflammatory cells. Hypothesis: A2AR activation during ECPR will improve survival and decrease the burden of injury in a large animal model of cardiac arrest. Methods: Adult swine underwent 20 minutes of circulatory arrest followed by defibrillation and 6 hours of ECPR. Animals were randomized to receive saline control (n=5) or the A2AR agonist Regadenoson (0.144 and 14.4mcg/kg/hr, n=5/group). Animals were subsequently weaned from ECPR and monitored for 24 hours. Clinical and biochemical endpoints were compared between groups. Results: The administration of Regadenoson increased survival after cardiac arrest compared to saline controls (10/10, 100% vs 2/5, 40%, p=0.02, Figure 1). Anesthetic administration (p=0.41), fluid resuscitation (p=0.54), and epinephrine required to maintain target arterial pressure (p=0.08) were similar for all subjects. Biochemical markers of organ damage, including creatinine (p=0.87), aspartate aminotransferase (p=0.89), and troponin I (p=0.38), were similar among groups (Figure 2). Conclusions: In a clinically relevant model of cardiac arrest treated with ECPR, selective A2AR agonism increased survival from 40% to 100% at 24 hours. These results suggest A2AR activation is a promising therapeutic target after cardiac arrest.

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Tetrathiomolybdate Treatment Attenuates Bleomycin-Induced Pulmonary Fibrosis in a Large Animal Model

10.1164/ajrccm-conference.2019.199.1_meetingabstracts.a2578 ◽

2019 ◽

Author(s):

H.B. Derseh ◽

K.U.E. Perera ◽

V.D.S. Nimanthi ◽

E. Koumoundouros ◽

A. Stent ◽

...

Keyword(s):

Animal Model ◽

Pulmonary Fibrosis ◽

Large Animal Model ◽

Large Animal

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Percutaneous Intramyocardial Septal Radiofrequency Ablation for Interventricular Septal Reduction: An Ovine Model with 1-Year Outcomes

Cardiology ◽

10.1159/000502973 ◽

2019 ◽

Vol 145 (1) ◽

pp. 53-62

Author(s):

Fang Liu ◽

Jianli Fu ◽

David Hsi ◽

Chao Sun ◽

Guangbin He ◽

...

Keyword(s):

Radiofrequency Ablation ◽

Sham Group ◽

Normal Sinus Rhythm ◽

Interventricular Septum ◽

Large Animal Model ◽

Lv Function ◽

Wall Thickening ◽

Large Animal ◽

Sheep Model

Background: Percutaneous intramyocardial (PIM) septal radiofrequency ablation (SRA) is a novel treatment approach for hypertrophic obstructive cardiomyopathy patients, but there has been lack of a large animal model to study PIM-SRA. We aimed to validate the long-term safety and efficacy of PIM-SRA and to observe pathological changes of the ablated interventricular septum (IVS) in a healthy sheep model. Methods and Results: Twelve sheep were randomized to the PIM-SRA group (n = 6) and the sham group (n = 6). In the PIM-SRA group, a radiofrequency (RF) electrode was inserted into the IVS with a maximum power of 80 W for 5 min. In the sham group, the RF electrode tip was positioned in the IVS segment but without RF power delivery. Septal hypokinesis was seen in all PIM-SRA group animals immediately after the procedure; the systolic wall thickening rate and motion amplitude of the ablated region decreased (p < 0.01), and the diastolic IVS thickness also decreased significantly over time (p < 0.01). ECG showed that all the sheep had normal sinus rhythm during the follow-up. Pathological examinations revealed scar tissue in the ablated region as expected. Conclusions: PIM-SRA produced precisely ablated myocardial tissue, reduced the IVS thickness significantly, preserved the global LV function, and avoided the incidence of conduction system damage in the long term. PIM-SRA was found to be a safe and effective minimally invasive septal reduction therapy.

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Nano-hemostats and a Pilot Study of Their Use in a Large Animal Model of Major Vessel Hemorrhage in Endoscopic Skull Base Surgery

Journal of Neurological Surgery Part B Skull Base ◽

10.1055/s-0036-1597277 ◽

2016 ◽

Vol 38 (03) ◽

pp. 215-221

Author(s):

Jae Murphy ◽

Sarah Vreugde ◽

Alkis Psaltis ◽

P. Wormald ◽

Alistair Jukes

Keyword(s):

Pilot Study ◽

Skull Base ◽

Skull Base Surgery ◽

Coagulation Cascade ◽

Large Animal Model ◽

Large Animal ◽

Small Pilot Study ◽

Sheep Model ◽

Endoscopic Skull Base Surgery ◽

Major Vessel

AbstractNano-hemostats are synthetic amino acid chains that self-assemble into a scaffold under certain conditions. These have been shown to be effective in stopping bleeding in small animal models of hemorrhage. Proposed mechanisms for their effect are that they form a mesh analogous to the fibrin plug in native hemostasis and that they may potentiate both platelet activation and the coagulation cascade. These may potentially become valuable adjuncts to endoscopic skull base surgery where there is the potential for both major vessel injury and smaller perforator injury to eloquent areas where bipolar cautery may not be suitable. We present a summary of the clinical studies to date and a small pilot study of nano-hemostat in an endoscopic sheep model of major vessel hemorrhage to determine its efficacy in stopping bleeding in this potentially catastrophic complication.

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Inhibition of the KCa3.1 Channel Alleviates Established Pulmonary Fibrosis in a Large Animal Model

American Journal of Respiratory Cell and Molecular Biology ◽

10.1165/rcmb.2016-0092oc ◽

2017 ◽

Vol 56 (4) ◽

pp. 539-550 ◽

Cited By ~ 10

Author(s):

Louise Organ ◽

Barbara Bacci ◽

Emmanuel Koumoundouros ◽

Wayne G. Kimpton ◽

Chrishan S. Samuel ◽

...

Keyword(s):

Animal Model ◽

Pulmonary Fibrosis ◽

Large Animal Model ◽

Large Animal

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Quantification of histomorphometric and structural bone changes in a sheep model for fracture treatment in osteoporotic bone

Veterinary and Comparative Orthopaedics and Traumatology ◽

10.1055/s-0038-1632790 ◽

2003 ◽

Vol 16 (04) ◽

pp. 243-249 ◽

Cited By ~ 1

Author(s):

E. Winterstein ◽

C. Eckhardt ◽

B. Rahn ◽

J. Goldhahn ◽

E. Schneider ◽

...

Keyword(s):

Iliac Crest ◽

Structural Parameters ◽

Large Animal Model ◽

Control Group ◽

Osteoporotic Bone ◽

Fracture Treatment ◽

Large Animal ◽

Average Dose ◽

Sheep Model ◽

Trabecular Thickness

SummaryThe histomorphometrical cancellous bone parameters of osteoporotic ewes were examined to establish a large animal model for the investigation of fracture treatment and healing in osteoporotic bone. Eighteen animals were divided into 2 groups. Group 1 (n = 8, age 3 to 5 years) served as an untreated control. In Group 2 (n = 10, age 7 to 9 years) osteoporosis was induced by means of ovariectomy, methylprednisolone application (average dose 160 mg/week) and Ca./Vit.D restricted diet during 7 months. Biopsies from iliac crest, femoral head and lumbar spine were harvested. Structural parameters from the biopsies were determined using μCT. In order to investigate static and dynamic parameters of bone remodelling, undecalcified sections were digitized and evaluated by means of image analysing software. Morphologically, bone area and trabecular thickness were reduced in osteoporotic sheep. Bone perimeter (−20%), trabecular number (−16%) and separation (+39%) showed a significant change in the iliac crest. The structure model index indicated a shift from the plate-to the rodmodel. At different skeletal sites of osteoporotic sheep bone resorption was significantly increased (250-675%), and bone formation significantly decreased (77-89%). Osteoid thickness (O.Th) was reduced (28-62%). In osteoporotic sheep the mean length of single labels was significantly shortened (64-73%) and the mineralising surface showed a significant decrease (80-92%). The mineralisation of the osteoid was comparable to the control group. The histomorphometrical changes in this sheep model of osteoporosis are comparable to the human situation of a combined post-menopausal and steroid-induced osteoporosis.

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KCa3.1 channel blockade attenuates microvascular remodelling in a large animal model of bleomycin-induced pulmonary fibrosis

Scientific Reports ◽

10.1038/s41598-019-56412-z ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 1

Author(s):

Habtamu B. Derseh ◽

Sasika N. Vithana Dewage ◽

Kopiyawaththage U. E. Perera ◽

Charles N. Pagel ◽

Emmanuel Koumoundouros ◽

...

Keyword(s):

Cell Proliferation ◽

Endothelial Cell ◽

Pulmonary Fibrosis ◽

Large Animal Model ◽

Endothelial Cell Proliferation ◽

Pulmonary Vasculature ◽

Large Animal ◽

Vascular Remodelling ◽

Vegf Expression ◽

Endothelial Growth Factor

AbstractIdiopathic pulmonary fibrosis (IPF) is a chronic progressive lung disease with limited therapeutic options and poor prognosis. IPF has been associated with aberrant vascular remodelling, however the role of vascular remodelling in pulmonary fibrosis is poorly understood. Here, we used a novel segmental challenge model of bleomycin-induced pulmonary fibrosis in sheep to evaluate the remodelling of the pulmonary vasculature, and to investigate the changes to this remodelling after the administration of the KCa3.1 channel inhibitor, senicapoc, compared to the FDA-approved drug pirfenidone. We demonstrate that in vehicle-treated sheep, bleomycin-infused lung segments had significantly higher blood vessel density when compared to saline-infused control segments in the same sheep. These microvascular density changes were significantly attenuated by senicapoc treatment. The increases in vascular endothelial growth factor (VEGF) expression and endothelial cell proliferation in bleomycin-infused lung segments were significantly reduced in sheep treated with the senicapoc, when compared to vehicle-treated controls. These parameters were not significantly suppressed with pirfenidone treatment. Senicapoc treatment attenuated vascular remodelling through inhibition of capillary endothelial cell proliferation and VEGF expression. These findings suggest a potential new mode of action for the novel drug senicapoc which may contribute to its efficacy in combatting pulmonary fibrosis.

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