Micro-computed tomography of pulmonary fibrosis in mice induced by adenoviral gene transfer of biologically active transforming growth factor-β1

Rodt, Thomas; Falck, Christian von; Dettmer, Sabine; Halter, Roman; Maus, Regina; Ask, Kjetil; Kolb, Martin; Gauldie, Jack; Hoy, Ludwig; Welte, Tobias; Galanski, Michael; Maus, Ulrich A.; Borlak, Jürgen

doi:10.1186/1465-9921-11-181

Cited by 35 publications

(31 citation statements)

References 26 publications

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“…Towards this end, we employed an adenoviral vector overexpressing TGF-β in mice. Adenoviral overexpression of TGF-β has induced lung fibrosis in monkeys [19] and wildtype mice [20,21] but not in Smad3 knockout mice [22]. In the current report, we subjected wildtype mice to lung injury by intratracheal administration of adenoviral vectors expressing the firefly gene-luciferase(Ad-Luc) or overexpressing TGF-β (Ad-TGF-β).…”

Section: Resultsmentioning

confidence: 99%

Potential Mechanisms Underlying TGF-β-mediated Complement Activation in Lung Fibrosis

Fisher¹,

Cipolla²,

Varre³

et al. 2017

Cell Mol Med

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While our previous studies suggest that limiting bleomycin-induced complement activation suppresses TGF-β signaling, the specific hierarchical interactions between TGF-β and complement in lung fibrosis are unclear. Herein, we investigated the mechanisms underlying TGF-β-induced complement activation in the pathogenesis of lung fibrosis. C57-BL6 mice were given intratracheal instillations of adenoviral vectors overexpressing TGF-β (Ad-TGFβ) or the firefly gene-luciferase (Ad-Luc; control). Two weeks later, mice with fibrotic lungs were instilled RNAi specific to receptors for C3a or C5a-C3ar or C5ar, and sacrificed at day 28. Histopathological analyses revealed that genetic silencing of C3ar or C5ar arrested the progression of TGF-β-induced lung fibrosis, collagen deposition and content (hydroxyproline, col1a1/2); and significantly suppressed local complement activation. With genetic silencing of either C3ar or C5ar, in Ad-TGFβ-injured lungs: we detected the recovery of Smad7 (TGF-β inhibitor) and diminished local release of DAF (membrane-bound complement inhibitor); in vitro: TGF-β-mediated loss of DAF was prevented. Conversely, blockade of the TGF-β receptor prevented C3a-mediated loss of DAF in both normal primary human alveolar and small airway epithelial cells. Of the 52 miRNAs analyzed as part of the Affymetrix array, normal primary human SAECs exposed to C3a, C5a or TGF-β caused discrete and overlapping miRNA regulation related to epithelial proliferation or apoptosis (miR-891A, miR-4442, miR-548, miR-4633), cellular contractility (miR-1197) and lung fibrosis . Our studies present potential mechanisms by which TGF-β activates complement and promotes lung fibrosis.

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Section: Resultsmentioning

confidence: 99%

Potential Mechanisms Underlying TGF-β-mediated Complement Activation in Lung Fibrosis

Fisher¹,

Cipolla²,

Varre³

et al. 2017

Cell Mol Med

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“…Few studies, however, have addressed the use of micro-CT for evaluation of the degree of fibrotic change in murine models of lung fibrosis. Previous studies [21][22][23] used in vivo micro-CT to investigate fibrotic changes in the adenoviral TGF-b1 overexpression-induced or bleomycin models of lung fibrosis in rats or mice with associated voxel sizes of ,155 mm, 94 mm or 35 mm. These studies used different approaches to segment the tissue into either fibrotic [21] or aerated lung volumes [22], or used semiquantitative pathological scoring [23], but the utility of these approaches has not yet been validated with a drug intervention strategy.…”

Section: Interstitial Lung Diseases | Cj Scotton Et Almentioning

confidence: 99%

“…Previous studies [21][22][23] used in vivo micro-CT to investigate fibrotic changes in the adenoviral TGF-b1 overexpression-induced or bleomycin models of lung fibrosis in rats or mice with associated voxel sizes of ,155 mm, 94 mm or 35 mm. These studies used different approaches to segment the tissue into either fibrotic [21] or aerated lung volumes [22], or used semiquantitative pathological scoring [23], but the utility of these approaches has not yet been validated with a drug intervention strategy. A recent study by JIN et al [24] used micro-CT (with ,44.25-mm spatial resolution) to investigate the therapeutic effects of rosiglitazone during BILF in mice; they reported beneficial changes in a qualitative radiological assessment, but did not provide a fully quantitative measure of fibrotic change.…”

Section: Interstitial Lung Diseases | Cj Scotton Et Almentioning

confidence: 99%

Ex vivomicro-computed tomography analysis of bleomycin-induced lung fibrosis for preclinical drug evaluation

et al. 2013

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Research into the pathogenesis underlying the development of idiopathic pulmonary fibrosis is hampered by a repertoire of animal models that fail to recapitulate all the features of the human disease. Better use and understanding of what the animal models represent may improve clinical predictability. We interrogated ex vivo micro-computed tomography (CT) as a novel end-point measure in the mouse model of bleomycin-induced lung fibrosis (BILF), and to evaluate a therapeutic dosing regimen for preclinical drug evaluation.A detailed characterisation of BILF was performed using standard end-point measures (lung hydroxyproline and histology). High resolution micro-CT (,13.7 mm voxel size) was evaluated for quantifying the extent and severity of lung fibrosis.The period from 14 to 28 days following bleomycin instillation represents progression of established fibrosis. A therapeutic dosing regimen during this period was validated using a transforming growth factor-b receptor-1 kinase inhibitor, and micro-CT provided a highly sensitive and quantitative measure of fibrosis. Moreover, fibrotic lesions did not completely resolve, but instead persisted for o6 months following a single insult with bleomycin.Ex vivo micro-CT analysis of BILF allows robust evaluation of therapeutic dosing once fibrosis is already well established, requiring fewer mice than conventional biochemical end-points. @ERSpublications Micro-CT has improved out understanding of the bleomycin model of fibrotic lung disease for preclinical drug evaluation

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“…24,25 The value of this imaging technique has been studied in disease models of lung fibrosis, lung emphysema and lung neoplasms. [26][27][28][29][30] Nevertheless, this technique has not been used for the assessment of invasive aspergillosis in small animals. Magnetic resonance imaging (MRI) provides images with good spatial resolution and excellent soft tissue contrast, thereby creating the potential to distinguish between different pathological processes occurring within the lung, such as inflammation or necrosis.…”

mentioning

confidence: 99%

Longitudinal, in vivo assessment of invasive pulmonary aspergillosis in mice by computed tomography and magnetic resonance imaging

Poelmans

Hillen

Vanherp

et al. 2016

Laboratory Investigation

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Invasive aspergillosis is an emerging threat to public health due to the increasing use of immune suppressive drugs and the emergence of resistance against antifungal drugs. To deal with this threat, research on experimental disease models provides insight into the pathogenesis of infections caused by susceptible and resistant Aspergillus strains and by assessing their response to antifungal drugs. However, standard techniques used to evaluate infection in a preclinical setting are severely limited by their invasive character, thereby precluding evaluation of disease extent and therapy effects in the same animal. To enable non-invasive, longitudinal monitoring of invasive pulmonary aspergillosis in mice, we optimized computed tomography (CT) and magnetic resonance imaging (MRI) techniques for daily follow-up of neutropenic BALB/c mice intranasally infected with A. fumigatus spores. Based on the images, lung parameters (signal intensity, lung tissue volume and total lung volume) were quantified to obtain objective information on disease onset, progression and extent for each animal individually. Fungal lung lesions present in infected animals were successfully visualized and quantified by both CT and MRI. By using an advanced MR pulse sequence with ultrashort echo times, pathological changes within the infected lung became visually and quantitatively detectable at earlier disease stages, thereby providing valuable information on disease onset and progression with high sensitivity. In conclusion, these noninvasive imaging techniques prove to be valuable tools for the longitudinal evaluation of dynamic disease-related changes and differences in disease severity in individual animals that might be readily applied for rapid and cost-efficient drug screening in preclinical models in vivo.

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Micro-computed tomography of pulmonary fibrosis in mice induced by adenoviral gene transfer of biologically active transforming growth factor-β1

Cited by 35 publications

References 26 publications

Potential Mechanisms Underlying TGF-β-mediated Complement Activation in Lung Fibrosis

Potential Mechanisms Underlying TGF-β-mediated Complement Activation in Lung Fibrosis

Ex vivomicro-computed tomography analysis of bleomycin-induced lung fibrosis for preclinical drug evaluation

Longitudinal, in vivo assessment of invasive pulmonary aspergillosis in mice by computed tomography and magnetic resonance imaging

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