Machine Learning Analysis of the Bleomycin Mouse Model Reveals the Compartmental and Temporal Inflammatory Pulmonary Fingerprint

Bordag, Natalie; Biasin, Valentina; Schnoegl, Diana; Valzano, Francesco; Jandl, Katharina; Nagy, Bence; Sharma, Neha; Wygrecka, Małgorzata; Kwapiszewska, Grażyna; Marsh, Leigh M.

doi:10.1016/j.isci.2020.101819

Cited by 21 publications

(20 citation statements)

References 40 publications

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“…Monocyte-derived macrophages, which contribute to the expanded CD11b + macrophages in our model, have been shown to be necessary for fibrogenesis ( 9 , 10 , 19 ). Using a model of concurrent mild bleomycin-induced lung injury, we assessed the impact of CS on macrophage function at two timepoints, day 7 (pre-tissue remodelling and early inflammation) and day 21 (peak fibrogenesis and late inflammation) ( Figure 5A ) ( 35 ). While bleomycin-treated mice had equivalent weight loss regardless of exposure ( Figure S5A ), 3/5 (60%) and 4/5 (80%) myeloid lineage-related genes were enriched in CS-exposed compared to RA-exposed bleomycin-treated mice at day 7 and 21 respectively ( Tables S3 and S4 ).…”

Section: Resultsmentioning

confidence: 99%

Increased Monocyte-Derived CD11b+ Macrophage Subpopulations Following Cigarette Smoke Exposure Are Associated With Impaired Bleomycin-Induced Tissue Remodelling

et al. 2021

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RationaleThe accumulation of macrophages in the airways and the pulmonary interstitium is a hallmark of cigarette smoke-associated inflammation. Notably, pulmonary macrophages are not a homogenous population but consist of several subpopulations. To date, the manner in which cigarette smoke exposure affects the relative composition and functional capacity of macrophage subpopulations has not been elucidated.MethodsUsing a whole-body cigarette smoke exposure system, we investigated the impact of cigarette smoke on macrophage subpopulations in C57BL/6 mice using flow cytometry-based approaches. Moreover, we used bromodeoxyuridine labelling plus Il1a-/- and Il1r1-/- mice to assess the relative contribution of local proliferation and monocyte recruitment to macrophage accumulation. To assess the functional consequences of altered macrophage subpopulations, we used a model of concurrent bleomycin-induced lung injury and cigarette smoke exposure to examine tissue remodelling processes.Main ResultsCigarette smoke exposure altered the composition of pulmonary macrophages increasing CD11b+ subpopulations including monocyte-derived alveolar macrophages (Mo-AM) as well as interstitial macrophages (IM)1, -2 and -3. The increase in CD11b+ subpopulations was observed at multiple cigarette smoke exposure timepoints. Bromodeoxyuridine labelling and studies in Il1a-/- mice demonstrated that increased Mo-AM and IM3 turnover in the lungs of cigarette smoke-exposed mice was IL-1α dependent. Compositional changes in macrophage subpopulations were associated with impaired induction of fibrogenesis including decreased α-smooth muscle actin positive cells following intratracheal bleomycin treatment. Mechanistically, in vivo and ex vivo assays demonstrated predominant macrophage M1 polarisation and reduced matrix metallopeptidase 9 activity in cigarette smoke-exposed mice.ConclusionCigarette smoke exposure modified the composition of pulmonary macrophage by expanding CD11b+ subpopulations. These compositional changes were associated with attenuated fibrogenesis, as well as predominant M1 polarisation and decreased fibrotic activity. Overall, these data suggest that cigarette smoke exposure altered the composition of pulmonary macrophage subpopulations contributing to impaired tissue remodelling.

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Section: Resultsmentioning

confidence: 99%

Increased Monocyte-Derived CD11b+ Macrophage Subpopulations Following Cigarette Smoke Exposure Are Associated With Impaired Bleomycin-Induced Tissue Remodelling

et al. 2021

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“…Data are presented as scatter dot plots with median. Dendrograms and heatmaps were created using R (version 3.6, R Core Team, Vienna, Austria) [38,43]. The dendrograms were clustered by Lance-Williams dissimilarity update with complete linkage (R function dist and hclust) and sorted (R function dendsort) at every merging point according to the average distance of subtrees and plotted at the corresponding heat maps (R function heatmap).…”

Section: Discussionmentioning

confidence: 99%

“…LSRII flow cytometer (BD, Biosciences, Vienna, Austria) and cytoFLEX-SII (Beckman Coulter, Vienna, Austria) were used to count the stained cells and FACSDiva software (BD, Biosciences, Vienna, Austria) and Flowjo v10 (LLC, Ashland, OR, USA) [37] for analysis. The distribution of the myeloid and lymphoid cells from the wild type animals of this study has been incorporated into a summary of our scientific work [38].…”

Section: Flow Cytometrymentioning

confidence: 99%

RGS5 Determines Neutrophil Migration in the Acute Inflammatory Phase of Bleomycin-Induced Lung Injury

Sharma

Nagaraj

Nagy

et al. 2021

IJMS

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The regulator of G protein signaling (RGS) represents a widespread system of controllers of cellular responses. The activities of the R4 subfamily of RGSs have been elucidated in allergic pulmonary diseases. However, the R4 signaling in other inflammatory lung diseases, with a strong cellular immune response, remained unexplored. Thus, our study aimed to discern the functional relevance of the R4 family member, RGS5, as a potential modulating element in this context. Gene profiling of the R4 subfamily showed increased RGS5 expression in human fibrosing lung disease samples. In line with this, RGS5 was markedly increased in murine lungs following bleomycin injury. RGS knock-out mice (RGS-/-) had preserved lung function while control mice showed significant combined ventilatory disorders three days after bleomycin application as compared to untreated control mice. Loss of RGS5 was associated with a significantly reduced neutrophil influx and tissue myeloperoxidase expression. In the LPS lung injury model, RGS5-/- mice also failed to recruit neutrophils into the lung, which was accompanied by reduced tissue myeloperoxidase levels after 24 h. Our in-vitro assays showed impaired migration of RGS5-/- neutrophils towards chemokines despite preserved Ca2+ signaling. ERK dephosphorylation might play a role in reduced neutrophil migration in our model. As a conclusion, loss of RGS5 preserves lung function and attenuates hyperinflammation in the acute phase of bleomycin-induced pulmonary fibrosis and LPS-induced lung injury. Targeting RGS5 might alleviate the severity of exacerbations in interstitial lung diseases.

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“…Bleomycin generates a well-established murine model of pneumonitis (20) with a dynamic pathology similar to that of COVID-19. Female, 11-13 week old C57BL/6 mice (Charles River Laboratories) were administered one intranasal 40 L dose of bleomycin sulphate (7.5 or 11.25 units/kg) or phosphate buffered saline (PBS) vehicle control under light isoflurane anaesthesia.…”

Section: Experimental Pneumonitismentioning

confidence: 99%

“…To address this need, we undertook preclinical studies using a mouse model of bleomycin-induced pneumonitis. Many pathophysiological changes of COVID-19 lung disease (epithelial cytopathy, endotheliitis, inflammatory infiltrates, surfactant loss) are reproduced in the pneumonitis induced by inhaled bleomycin (19,20). Indeed, single cell sequencing studies of mouse bleomycin (21) and COVID-19 (22,23) pneumonitis have shown pathogenic SPP1 pos macrophage orthologues expressing key inflammatory mediators to be prominent in both conditions, along with profoundly reduced expression of anticoagulant and anti-apoptotic protein S in alveolar macrophages.…”

Section: Introductionmentioning

confidence: 99%

Low-dose lung radiotherapy for COVID-19 lung disease: a preclinical efficacy study in a bleomycin model of pneumonitis

Jackson

Stevenson

Chahal

et al. 2021

Preprint

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PurposeLow-dose whole lung radiotherapy (LDLR) has been proposed as a treatment for patients with acute respiratory distress syndrome associated with SARS-CoV-2 infection and clinical trials are underway. There is an urgent need for preclinical evidence to justify this approach and inform dose, scheduling and mechanisms of action.Materials and methodsFemale C57BL/6 mice were treated with intranasal bleomycin sulphate (7.5 or 11.25 units/kg, day 0), then exposed to whole lung radiation therapy (0.5, 1.0, 1.5 Gy or sham, day 3). Bodyweight was measured daily and lung tissue harvested for histology and flow cytometry on day 10. Computed tomography (CT) lung imaging was performed pre-radiation (day 3) and pre-endpoint (day 10).ResultsBleomycin caused pneumonitis of variable severity which correlated with weight loss. LDLR at 1.0 Gy was associated with a significant increase in the proportion of mice recovering to 98% of initial bodyweight and a proportion of these mice exhibited less severe histopathological lung changes. Mice experiencing moderate initial weight loss were more likely to respond to LDLR than those experiencing severe initial weight loss. Additionally, LDLR (1.0 Gy) significantly reduced bleomycin-induced increases in interstitial macrophages, CD103+ dendritic cells and neutrophil-DC hybrids. Overall,bleomycin-treated mice exhibited significantly higher percentages of non-aerated lung in left than right lungs and LDLR (1.0 Gy) prevented further reductions in aerated lung volume in right but not left lungs. LDLR at 0.5 and 1.5 Gy did not modulate bodyweight or flow cytometric readouts of bleomycin-induced pneumonitis.ConclusionsOur data support the concept that LDLR can ameliorate acute inflammatory lung injury, identify 1.0 Gy as the most effective dose and provide preliminary evidence that it is more effective in the context of moderate than severe pneumonitis. Mechanistically, LDLR at 1.0 Gy significantly suppressed bleomycin-induced accumulation of pulmonary interstitial macrophages, CD103+ dendritic cells and neutrophil-DC hybrids.

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Machine Learning Analysis of the Bleomycin Mouse Model Reveals the Compartmental and Temporal Inflammatory Pulmonary Fingerprint

Cited by 21 publications

References 40 publications

Increased Monocyte-Derived CD11b+ Macrophage Subpopulations Following Cigarette Smoke Exposure Are Associated With Impaired Bleomycin-Induced Tissue Remodelling

Increased Monocyte-Derived CD11b+ Macrophage Subpopulations Following Cigarette Smoke Exposure Are Associated With Impaired Bleomycin-Induced Tissue Remodelling

RGS5 Determines Neutrophil Migration in the Acute Inflammatory Phase of Bleomycin-Induced Lung Injury

Low-dose lung radiotherapy for COVID-19 lung disease: a preclinical efficacy study in a bleomycin model of pneumonitis

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