Pub Date : 2018-09-15DOI: 10.1183/13993003.CONGRESS-2018.PA1386
Ayako Miura, S. Yanagi, Hironobu Tsubouchi, A. Matsuo, N. Matsumoto, M. Nakazato
Introduction: Cellular senescence (CS) is a crucial anticancer mechanism that is defined by irreversible cell-cycle arrest and senescence-associated secretory phenotype (SASP). CS also participates in tissue remodeling during normal embryonic development (i.e., programmed CS): however, the roles of programmed CS in lung development and their intrinsic regulator(s) remain unknown. Aims: To determine the role of epithelial Pten, a tumor suppressor, in programmed CS and cell-fate decision during lung development. Methods: We conducted structural and biochemical analyses of two different types of lung epithelium-specific Pten-deleted mice (ShhGFPCre/PtenΔ/Δ mice and SPC-rtTA/(tetO)7Cre/PtenΔ/Δ; SOPtenΔ/Δ mice). Further mechanistic studies were performed in vitro and in vivo. Results: Both ShhGFPCre/PtenΔ/Δ and SOPtenΔ/Δ mice showed impaired development of mature alveoli. A microarray study identified attenuated expressions of alveolar type 2 markers whereas detected increased expressions of markers of neuroendocrine cells, Club cells, goblet cells, and alveolar type 1 in lung epithelial cells isolated from SOPtenΔ/Δ mice. Deletion of Pten in lung epithelium enhances NF-κB–Notch signaling pathway. Pten deficiency led to cell growth arrest of lung epithelial cells, increased number of senescent cells, and enhanced SASP at E18.5. Epithelial Pten ablation upregulated p53–p21 pathway and impaired bipolar spindle formation in vivo and in vitro. Conclusions: Our results demonstrate that epithelial Pten has an essential role in controlling programmed CS and cell-fate choices during lung development by regulating NF-κB–Notch pathway, p53–p21 pathway, and proper architectural integrity of the mitotic spindle.
{"title":"The role of epithelial Pten in programmed cellular senescence during lung development","authors":"Ayako Miura, S. Yanagi, Hironobu Tsubouchi, A. Matsuo, N. Matsumoto, M. Nakazato","doi":"10.1183/13993003.CONGRESS-2018.PA1386","DOIUrl":"https://doi.org/10.1183/13993003.CONGRESS-2018.PA1386","url":null,"abstract":"Introduction: Cellular senescence (CS) is a crucial anticancer mechanism that is defined by irreversible cell-cycle arrest and senescence-associated secretory phenotype (SASP). CS also participates in tissue remodeling during normal embryonic development (i.e., programmed CS): however, the roles of programmed CS in lung development and their intrinsic regulator(s) remain unknown. Aims: To determine the role of epithelial Pten, a tumor suppressor, in programmed CS and cell-fate decision during lung development. Methods: We conducted structural and biochemical analyses of two different types of lung epithelium-specific Pten-deleted mice (ShhGFPCre/PtenΔ/Δ mice and SPC-rtTA/(tetO)7Cre/PtenΔ/Δ; SOPtenΔ/Δ mice). Further mechanistic studies were performed in vitro and in vivo. Results: Both ShhGFPCre/PtenΔ/Δ and SOPtenΔ/Δ mice showed impaired development of mature alveoli. A microarray study identified attenuated expressions of alveolar type 2 markers whereas detected increased expressions of markers of neuroendocrine cells, Club cells, goblet cells, and alveolar type 1 in lung epithelial cells isolated from SOPtenΔ/Δ mice. Deletion of Pten in lung epithelium enhances NF-κB–Notch signaling pathway. Pten deficiency led to cell growth arrest of lung epithelial cells, increased number of senescent cells, and enhanced SASP at E18.5. Epithelial Pten ablation upregulated p53–p21 pathway and impaired bipolar spindle formation in vivo and in vitro. Conclusions: Our results demonstrate that epithelial Pten has an essential role in controlling programmed CS and cell-fate choices during lung development by regulating NF-κB–Notch pathway, p53–p21 pathway, and proper architectural integrity of the mitotic spindle.","PeriodicalId":290970,"journal":{"name":"Lung and airway developmental biology","volume":"86 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2018-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"114509558","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2018-09-15DOI: 10.1183/13993003.CONGRESS-2018.PA1378
S. Danopoulos, S. Bhattacharya, M. Thornton, B. Grubbs, T. Mariani, D. A. Alam
The mesenchyme gives rise to multiple distinct cell lineages in the mature respiratory system, including smooth muscle cells of the airway and vasculature, vascular endothelial cells, and parenchymal fibroblasts. However, a thorough understanding of the specification and the inter-relationships among the diverse mesenchymally derived cells in the human lung is lacking. We used single cell RNAseq of human fetal lung cells, to characterize cellular phenotypes in the pseudoglandular (11.5 weeks) and early canalicular (18.5 weeks) stage human lung. Confirmed live cells obtained from protease-dispersed tissue were captured and sequenced using the Chromium 10X system. Expression analysis was performed using Seurat 2.0 and functional analysis was performed using ToppFun. At these early stages, we were able to identify molecularly distinct cell populations representing fetal human lung endothelial cells, pericytes and smooth muscle cells. Early endothelial lineages expressed “classic” endothelial cell markers (PECAM, CDH5, VWF) as well as CD34, KIT and CLDN5. Cells with pericyte characteristics were evident early in development, and defined by expression of PDGFRB, THY1, notch signaling, and broad expression of basement membrane molecules (COL4, laminin, proteoglycans). Smooth muscle cells, defined by expression of the canonical lineage marker ACTA2, demonstrated evidence for Hedgehog signaling, with high expression of HHIP and PTCH1. At both 11 and 18 weeks, we identified a large population of undefined mesenchymal cells characterized by expression of COL1, PDGFRA and elastin fiber genes (ELN, MFAP4, FBLN1, LOXL1). Our data suggest that early specification of distinct mesenchymal lineages occurs in the human lung.
{"title":"Diversity in mesenchymal lineages during early human lung development","authors":"S. Danopoulos, S. Bhattacharya, M. Thornton, B. Grubbs, T. Mariani, D. A. Alam","doi":"10.1183/13993003.CONGRESS-2018.PA1378","DOIUrl":"https://doi.org/10.1183/13993003.CONGRESS-2018.PA1378","url":null,"abstract":"The mesenchyme gives rise to multiple distinct cell lineages in the mature respiratory system, including smooth muscle cells of the airway and vasculature, vascular endothelial cells, and parenchymal fibroblasts. However, a thorough understanding of the specification and the inter-relationships among the diverse mesenchymally derived cells in the human lung is lacking. We used single cell RNAseq of human fetal lung cells, to characterize cellular phenotypes in the pseudoglandular (11.5 weeks) and early canalicular (18.5 weeks) stage human lung. Confirmed live cells obtained from protease-dispersed tissue were captured and sequenced using the Chromium 10X system. Expression analysis was performed using Seurat 2.0 and functional analysis was performed using ToppFun. At these early stages, we were able to identify molecularly distinct cell populations representing fetal human lung endothelial cells, pericytes and smooth muscle cells. Early endothelial lineages expressed “classic” endothelial cell markers (PECAM, CDH5, VWF) as well as CD34, KIT and CLDN5. Cells with pericyte characteristics were evident early in development, and defined by expression of PDGFRB, THY1, notch signaling, and broad expression of basement membrane molecules (COL4, laminin, proteoglycans). Smooth muscle cells, defined by expression of the canonical lineage marker ACTA2, demonstrated evidence for Hedgehog signaling, with high expression of HHIP and PTCH1. At both 11 and 18 weeks, we identified a large population of undefined mesenchymal cells characterized by expression of COL1, PDGFRA and elastin fiber genes (ELN, MFAP4, FBLN1, LOXL1). Our data suggest that early specification of distinct mesenchymal lineages occurs in the human lung.","PeriodicalId":290970,"journal":{"name":"Lung and airway developmental biology","volume":"21 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2018-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"126242591","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2018-09-15DOI: 10.1183/13993003.CONGRESS-2018.PA1376
M. Kovach, Ulrika Käck, K. Che, Bettina Levänen, G. Lilja, J. Konradsen, A. Lindén
{"title":"Late Breaking Abstract - Systemic IL-26 correlates with improved asthma control in children with allergic sensitization","authors":"M. Kovach, Ulrika Käck, K. Che, Bettina Levänen, G. Lilja, J. Konradsen, A. Lindén","doi":"10.1183/13993003.CONGRESS-2018.PA1376","DOIUrl":"https://doi.org/10.1183/13993003.CONGRESS-2018.PA1376","url":null,"abstract":"","PeriodicalId":290970,"journal":{"name":"Lung and airway developmental biology","volume":"1 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2018-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"129658831","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2018-09-15DOI: 10.1183/13993003.CONGRESS-2018.LSC-1093
J. Collins, M. Lithopoulos, C. D. Santos, N. Issa, M. Moebius, C. Ito, S. Zhong, A. Vadivel, B. Thébaud
Background: Bronchopulmonary dysplasia (BPD), a common adverse outcome of extreme preterm birth, can be caused by oxygen-related lung injury and is characterized by arrested alveolar development. Mesenchymal stromal cells (MSCs) have lung protective effects. Conversely, BPD is associated with increased MSCs in tracheal aspirates. This apparent discrepancy is unexplored. We hypothesized that endogenous lung (L-)MSCs are perturbed in an oxygen-induced rat model mimicking BPD. Methods: Rat pups were exposed to 21% or 95% O2 from postnatal day 0 to 10. On day 12, CD146+ L-MSCs were isolated and characterized. Epithelial and vascular repair potential were tested by scratch assay and endothelial network formation respectively, immune function by mixed lymphocyte reaction assay. Microarray analysis was performed using GSEA software. Results: L-MSCs isolated from hyperoxia rat pups had decreased CD73 expression and inhibited lung endothelial network formation. L-MSCs indiscriminately promoted epithelial wound healing and limited T-cell proliferation. Expression of anti-angiogenic genes of the axonal guidance cue pathway was increased after in vivo hyperoxia, whereas genes of the anti-inflammatory JAK/STAT and lung/vascular growth promoting FGF pathways were decreased. Conclusions: In vivo hyperoxia exposure alters the pro-angiogenic effects and FGF expression of L-MSCs. Additionally, decreased CD73 and JAK/STAT expression suggest decreased immune function. L-MSC function may be perturbed and contribute to BPD pathogenesis. These findings may lead to improvements in manufacturing exogenous MSCs with superior repair capabilities.
{"title":"Impaired angiogenic supportive capacity and altered gene expression profile of CD146+ mesenchymal stromal cells isolated from hyperoxia-injured neonatal rat lungs","authors":"J. Collins, M. Lithopoulos, C. D. Santos, N. Issa, M. Moebius, C. Ito, S. Zhong, A. Vadivel, B. Thébaud","doi":"10.1183/13993003.CONGRESS-2018.LSC-1093","DOIUrl":"https://doi.org/10.1183/13993003.CONGRESS-2018.LSC-1093","url":null,"abstract":"Background: Bronchopulmonary dysplasia (BPD), a common adverse outcome of extreme preterm birth, can be caused by oxygen-related lung injury and is characterized by arrested alveolar development. Mesenchymal stromal cells (MSCs) have lung protective effects. Conversely, BPD is associated with increased MSCs in tracheal aspirates. This apparent discrepancy is unexplored. We hypothesized that endogenous lung (L-)MSCs are perturbed in an oxygen-induced rat model mimicking BPD. Methods: Rat pups were exposed to 21% or 95% O2 from postnatal day 0 to 10. On day 12, CD146+ L-MSCs were isolated and characterized. Epithelial and vascular repair potential were tested by scratch assay and endothelial network formation respectively, immune function by mixed lymphocyte reaction assay. Microarray analysis was performed using GSEA software. Results: L-MSCs isolated from hyperoxia rat pups had decreased CD73 expression and inhibited lung endothelial network formation. L-MSCs indiscriminately promoted epithelial wound healing and limited T-cell proliferation. Expression of anti-angiogenic genes of the axonal guidance cue pathway was increased after in vivo hyperoxia, whereas genes of the anti-inflammatory JAK/STAT and lung/vascular growth promoting FGF pathways were decreased. Conclusions: In vivo hyperoxia exposure alters the pro-angiogenic effects and FGF expression of L-MSCs. Additionally, decreased CD73 and JAK/STAT expression suggest decreased immune function. L-MSC function may be perturbed and contribute to BPD pathogenesis. These findings may lead to improvements in manufacturing exogenous MSCs with superior repair capabilities.","PeriodicalId":290970,"journal":{"name":"Lung and airway developmental biology","volume":"37 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2018-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"124984904","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2018-09-15DOI: 10.1183/13993003.CONGRESS-2018.PA1383
E. Eenjes, M. B. Kempen, A. Munck, Anneloes Van Krimpen, L. Bruin, R. Rottier
{"title":"Sox21, a novel player in the proximal-distal specification of airway epithelium","authors":"E. Eenjes, M. B. Kempen, A. Munck, Anneloes Van Krimpen, L. Bruin, R. Rottier","doi":"10.1183/13993003.CONGRESS-2018.PA1383","DOIUrl":"https://doi.org/10.1183/13993003.CONGRESS-2018.PA1383","url":null,"abstract":"","PeriodicalId":290970,"journal":{"name":"Lung and airway developmental biology","volume":"229 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2018-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"123310060","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2018-09-15DOI: 10.1183/13993003.CONGRESS-2018.PA1380
A. Pincus, A. Fryer, D. Jacoby, K. Lebold
Background: Maternal asthma causes airway hyperreactivity in infants at birth and increases asthma risk in childhood. Humans with severe asthma have increased sensory nerve density in airways. Aim: We investigated whether maternal allergen exposure affects airway reactivity and innervation in their adult offspring. Methods: Female C57BI/6 mice were exposed to house dust mite (HDM) or vehicle (VEH) daily for 4 weeks prior to pregnancy and throughout gestation. At 8 weeks of age, offspring were sensitized and challenged with HDM. 24 hours after final challenge, mice were anesthetized and ventilated, and changes in serotonin-induced bronchoconstriction were measured. Inflammatory cells, Th2 cytokines, and neurotrophins were measured in bronchoalveolar lavage (BAL). Sensory nerve length and branching was quantified in airway epithelium. Results: Following HDM sensitization and challenge, offspring from HDM exposed mothers had significantly increased airway hyperreactivity, eosinophils and macrophages compared with offspring from VEH exposed mothers. IL-5 and IL-13 were not different in BAL of offspring from HDM vs VEH exposed mothers. Offspring from HDM exposed mothers had increased sensory nerve length and branching in airway epithelium and increased brain-derived neurotrophic factor compared with offspring from VEH exposed mothers. Conclusions: Exposure to maternal antigen in utero increases sensory innervation and neurotrophin expression in airways of their offspring. Subsequent exposure of these offspring to antigen results in significantly greater airway hyperreactivity, and macrophage and eosinophilic inflammation. NIH grant support: HL132414, HL124165, HL065228, HL083808
{"title":"Maternal allergen exposure increases offspring sensory airway innervation and airway hyperreactivity in mice","authors":"A. Pincus, A. Fryer, D. Jacoby, K. Lebold","doi":"10.1183/13993003.CONGRESS-2018.PA1380","DOIUrl":"https://doi.org/10.1183/13993003.CONGRESS-2018.PA1380","url":null,"abstract":"Background: Maternal asthma causes airway hyperreactivity in infants at birth and increases asthma risk in childhood. Humans with severe asthma have increased sensory nerve density in airways. Aim: We investigated whether maternal allergen exposure affects airway reactivity and innervation in their adult offspring. Methods: Female C57BI/6 mice were exposed to house dust mite (HDM) or vehicle (VEH) daily for 4 weeks prior to pregnancy and throughout gestation. At 8 weeks of age, offspring were sensitized and challenged with HDM. 24 hours after final challenge, mice were anesthetized and ventilated, and changes in serotonin-induced bronchoconstriction were measured. Inflammatory cells, Th2 cytokines, and neurotrophins were measured in bronchoalveolar lavage (BAL). Sensory nerve length and branching was quantified in airway epithelium. Results: Following HDM sensitization and challenge, offspring from HDM exposed mothers had significantly increased airway hyperreactivity, eosinophils and macrophages compared with offspring from VEH exposed mothers. IL-5 and IL-13 were not different in BAL of offspring from HDM vs VEH exposed mothers. Offspring from HDM exposed mothers had increased sensory nerve length and branching in airway epithelium and increased brain-derived neurotrophic factor compared with offspring from VEH exposed mothers. Conclusions: Exposure to maternal antigen in utero increases sensory innervation and neurotrophin expression in airways of their offspring. Subsequent exposure of these offspring to antigen results in significantly greater airway hyperreactivity, and macrophage and eosinophilic inflammation. NIH grant support: HL132414, HL124165, HL065228, HL083808","PeriodicalId":290970,"journal":{"name":"Lung and airway developmental biology","volume":"24 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2018-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"114424442","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2018-09-15DOI: 10.1183/13993003.CONGRESS-2018.PA1379
D. Hirani, M. Koch, K. Dinger, J. Mohr, C. Vohlen, C. Klaudt, J. Dötsch, M. A. Alcázar
Background: Bronchopulmonary dysplasia (BPD), a chronic lung disease of premature infants, is characterized by arrest of alveolarization and matrix remodeling. Myofibroblasts are crucial in both processes. Since the transcription factor Kruppel-like factor 4 (Klf4) regulates cell homeostasis and fibroblast function, we studied the functional role of Klf4 in lungs of newborn mice exposed to hyperoxia and in primary neonatal lung fibroblasts (PnF). Methods: (1) Newborn mice were exposed to 85% O2 (HYX) or 21% O2 (NOX) for up to 28 days. (2) Knockdown (siRNA) or overexpression (sleeping beauty transposon system) of Klf4 was induced in PnF or mouse embryonic fibroblasts (MEFs), respectively, followed by exposure to HYX or NOX for up to 48 hours. Results: (1) HYX markedly reduced Klf4 mRNA and protein expression at P7 and P28 in lungs. Gene expression of TGFβ, PAI-1 and αSMA protein (indicator of myofibroblasts) were higher in lungs after HYX. Immunostaining showed a localization of Klf4 in myofibroblasts (αSMA-positive cells). (2) Exposure of PnF to HYX decreased Klf4 protein. Both Knockdown of Klf4 and HYX reduced migration and increased mRNA of CTGF, collagen Iα1 & IVα1 and PDGFRα. Proliferation was not affected by loss of Klf4. In contrast, overexpression of Klf4 in MEFs confirmed the modulation of migration and ECM expression by Klf4 and abrogated the changes induced by HYX. Conclusion: We identify Klf4 as a novel key regulator of neonatal lung fibroblast homeostasis. Loss of Klf4 is intimately linked to myofibroblast activation, fibrosis and migration, eventually reducing thereby alveolar formation and contributing to the pathogeneis of BPD.
{"title":"Krüppel-like factor 4 (Klf4) is a novel regulator of neonatal lung fibroblast homeostasis and reduced in hyperoxia-induced lung injury","authors":"D. Hirani, M. Koch, K. Dinger, J. Mohr, C. Vohlen, C. Klaudt, J. Dötsch, M. A. Alcázar","doi":"10.1183/13993003.CONGRESS-2018.PA1379","DOIUrl":"https://doi.org/10.1183/13993003.CONGRESS-2018.PA1379","url":null,"abstract":"Background: Bronchopulmonary dysplasia (BPD), a chronic lung disease of premature infants, is characterized by arrest of alveolarization and matrix remodeling. Myofibroblasts are crucial in both processes. Since the transcription factor Kruppel-like factor 4 (Klf4) regulates cell homeostasis and fibroblast function, we studied the functional role of Klf4 in lungs of newborn mice exposed to hyperoxia and in primary neonatal lung fibroblasts (PnF). Methods: (1) Newborn mice were exposed to 85% O2 (HYX) or 21% O2 (NOX) for up to 28 days. (2) Knockdown (siRNA) or overexpression (sleeping beauty transposon system) of Klf4 was induced in PnF or mouse embryonic fibroblasts (MEFs), respectively, followed by exposure to HYX or NOX for up to 48 hours. Results: (1) HYX markedly reduced Klf4 mRNA and protein expression at P7 and P28 in lungs. Gene expression of TGFβ, PAI-1 and αSMA protein (indicator of myofibroblasts) were higher in lungs after HYX. Immunostaining showed a localization of Klf4 in myofibroblasts (αSMA-positive cells). (2) Exposure of PnF to HYX decreased Klf4 protein. Both Knockdown of Klf4 and HYX reduced migration and increased mRNA of CTGF, collagen Iα1 & IVα1 and PDGFRα. Proliferation was not affected by loss of Klf4. In contrast, overexpression of Klf4 in MEFs confirmed the modulation of migration and ECM expression by Klf4 and abrogated the changes induced by HYX. Conclusion: We identify Klf4 as a novel key regulator of neonatal lung fibroblast homeostasis. Loss of Klf4 is intimately linked to myofibroblast activation, fibrosis and migration, eventually reducing thereby alveolar formation and contributing to the pathogeneis of BPD.","PeriodicalId":290970,"journal":{"name":"Lung and airway developmental biology","volume":"213 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2018-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"124194833","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2018-09-15DOI: 10.1183/13993003.CONGRESS-2018.PA1388
Kimberley C. W. Wang, J. Elliot, S. Saglani, P. Noble, A. James
Background: The volume of the airway smooth muscle (ASM) layer increases from birth until adulthood. However it is unknown if the increased volume is due to ASM cell hypertrophy and/or hyperplasia, or an increase in the proportion of extracellular matrix (ECM). It is important to investigate the mechanisms that produce the normal increase in the volume of the ASM layer as these may have different effects on airway function and/or be susceptible to change resulting in abnormal function. Aim: Characterise the structural mechanisms producing growth of the ASM layer in early life. Methods: Airways from post-mortem cases were available from 4 different age groups: Results: Airway size and muscle thickness approximately doubled from the prenatal period to the first year of life. The increase in ASM layer thickness was accompanied by a reduction in ASM cell density. There was a non-significant trend for an increase in NL. Conclusion: From late gestation to the first year of life, the ASM layer thickens with a concomitant reduction in ASM cell density. This may due to cell hypertrophy or a disproportionate expansion of the ECM between cells. There is a suggestion of increased cell number between 6–12 months of age.
{"title":"Thickening of the airway smooth muscle layer from late gestation to first year of life is accompanied by a reduction in smooth muscle cell density","authors":"Kimberley C. W. Wang, J. Elliot, S. Saglani, P. Noble, A. James","doi":"10.1183/13993003.CONGRESS-2018.PA1388","DOIUrl":"https://doi.org/10.1183/13993003.CONGRESS-2018.PA1388","url":null,"abstract":"Background: The volume of the airway smooth muscle (ASM) layer increases from birth until adulthood. However it is unknown if the increased volume is due to ASM cell hypertrophy and/or hyperplasia, or an increase in the proportion of extracellular matrix (ECM). It is important to investigate the mechanisms that produce the normal increase in the volume of the ASM layer as these may have different effects on airway function and/or be susceptible to change resulting in abnormal function. Aim: Characterise the structural mechanisms producing growth of the ASM layer in early life. Methods: Airways from post-mortem cases were available from 4 different age groups: Results: Airway size and muscle thickness approximately doubled from the prenatal period to the first year of life. The increase in ASM layer thickness was accompanied by a reduction in ASM cell density. There was a non-significant trend for an increase in NL. Conclusion: From late gestation to the first year of life, the ASM layer thickens with a concomitant reduction in ASM cell density. This may due to cell hypertrophy or a disproportionate expansion of the ECM between cells. There is a suggestion of increased cell number between 6–12 months of age.","PeriodicalId":290970,"journal":{"name":"Lung and airway developmental biology","volume":"24 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2018-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"121667512","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2018-09-15DOI: 10.1183/13993003.CONGRESS-2018.PA1389
M. Ljujić, Máté Varga, S. Bartel, S. Krauss‐Etschmann, A. D. Rankov
There is an evident need for new animal models to understand molecular mechanisms underlying development of chronic lung diseases, such as asthma and COPD. Recently, zebrafish (Danio rerio) has emerged as a model for various aspects of lung disease (inflammation, acute lung injury and effects of cigarette smoke). The overall aim is to establish the zebrafish as a model to study developmental origins of chronic lung disease and their transgenerational inheritance. The objective of this study was to investigate effects of nicotine on zebrafish development, selected gene expression and neutrophil chemotaxis in order to validate its usefulness as a transgenerational model. Zebrafish Tubingen wild type and Tg(mpx:GFP), were maintained and bread according to ZFbook. Toxicity of different nicotine concentrations was assessed by the analysis of survival, hatching and malformations. Real-time PCR was used to analyse expression of CYP1A1 and AHR1 genes and neutrophil counts and migration were analysed and recorded by live imaging. There was increased mortality in groups treated with 20µg/mL nicotine. An increase in malformations was present from concentration of 10µg/mL (pericardial oedema, body curvature, shortening of body and decreased movements and swim bladder inflation). The expression of CYP1A1 was observed to be increased. There was an increase of number of neutrophils and they migrated from circulation after treatment with nicotine. There is a correlation between the effects of nicotine on mammalian organisms and zebrafish. These results indicate usefulness zebrafish as a model for various aspects of chronic lung diseases. In future studies we will analyse the effect of nicotine on subsequent generations.
{"title":"Zebrafish as a model for study of developmental origins of chronic lung diseases","authors":"M. Ljujić, Máté Varga, S. Bartel, S. Krauss‐Etschmann, A. D. Rankov","doi":"10.1183/13993003.CONGRESS-2018.PA1389","DOIUrl":"https://doi.org/10.1183/13993003.CONGRESS-2018.PA1389","url":null,"abstract":"There is an evident need for new animal models to understand molecular mechanisms underlying development of chronic lung diseases, such as asthma and COPD. Recently, zebrafish (Danio rerio) has emerged as a model for various aspects of lung disease (inflammation, acute lung injury and effects of cigarette smoke). The overall aim is to establish the zebrafish as a model to study developmental origins of chronic lung disease and their transgenerational inheritance. The objective of this study was to investigate effects of nicotine on zebrafish development, selected gene expression and neutrophil chemotaxis in order to validate its usefulness as a transgenerational model. Zebrafish Tubingen wild type and Tg(mpx:GFP), were maintained and bread according to ZFbook. Toxicity of different nicotine concentrations was assessed by the analysis of survival, hatching and malformations. Real-time PCR was used to analyse expression of CYP1A1 and AHR1 genes and neutrophil counts and migration were analysed and recorded by live imaging. There was increased mortality in groups treated with 20µg/mL nicotine. An increase in malformations was present from concentration of 10µg/mL (pericardial oedema, body curvature, shortening of body and decreased movements and swim bladder inflation). The expression of CYP1A1 was observed to be increased. There was an increase of number of neutrophils and they migrated from circulation after treatment with nicotine. There is a correlation between the effects of nicotine on mammalian organisms and zebrafish. These results indicate usefulness zebrafish as a model for various aspects of chronic lung diseases. In future studies we will analyse the effect of nicotine on subsequent generations.","PeriodicalId":290970,"journal":{"name":"Lung and airway developmental biology","volume":"25 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2018-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"117057581","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2018-09-15DOI: 10.1183/13993003.CONGRESS-2018.PA1387
I. Katovich, Zhanna Rutkouskaya, S. Anischenko, A. Tahanovich
Lung-protective pharmacotherapies preventing development of bronchopulmonary dysplasia (BPD) in infants are widely investigated. The aim of this study was to evaluate the influence of inhaled N-acetylcysteine (NAC) on the oxidative and protease imbalance in lungs of newborn guinea pigs under prolonged hyperoxia. The studied groups included: “hyperoxia” (n=12, 70% O2, 14 days), “hyperoxia+NAC” (n=6, 250 mg/kg of NAC introduced by nebulizer on alternate days on hyperoxia exposure), “control” (n=12, room air). Hyperoxia exposure resulted in enhanced production of reactive oxygen species (ROS) by bronchoalveolar lavage cells, 1,9-fold increased level of products reacting with thiobarbituric acid (TBARP), low glutathione peroxidase activity and glutathione (G-SH) in bronchoalveolar lavage fluid (BALF) (30% and 68% of controls respectively); elevated alpha1-antitrypsin (A1-AT), neutrophil elastase and matrix metalloproteases MMP2 and MMP9 in lung homogenates (141%, 286%, 132%, and 176% of controls respectively) (for all p
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