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1.
Am J Physiol Lung Cell Mol Physiol ; 303(12): L1046-56, 2012 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-23043074

RESUMO

Idiopathic pulmonary fibrosis (IPF) is a progressive fibrotic lung disease without effective therapeutics. Periostin has been reported to be elevated in IPF patients relative to controls, but its sources and mechanisms of action remain unclear. We confirm excess periostin in lungs of IPF patients and show that IPF fibroblasts produce periostin. Blood was obtained from 54 IPF patients (all but 1 with 48 wk of follow-up). We show that periostin levels predict clinical progression at 48 wk (hazard ratio = 1.47, 95% confidence interval = 1.03-2.10, P < 0.05). Monocytes and fibrocytes are sources of periostin in circulation in IPF patients. Previous studies suggest that periostin may regulate the inflammatory phase of bleomycin-induced lung injury, but periostin effects during the fibroproliferative phase of the disease are unknown. Wild-type and periostin-deficient (periostin(-/-)) mice were anesthetized and challenged with bleomycin. Wild-type mice were injected with bleomycin and then treated with OC-20 Ab (which blocks periostin and integrin interactions) or control Ab during the fibroproliferative phase of disease, and fibrosis and survival were assessed. Periostin expression was upregulated quickly after treatment with bleomycin and remained elevated. Periostin(-/-) mice were protected from bleomycin-induced fibrosis. Instillation of OC-20 during the fibroproliferative phase improved survival and limited collagen deposition. Chimeric mouse studies suggest that hematopoietic and structural sources of periostin contribute to lung fibrogenesis. Periostin was upregulated by transforming growth factor-ß in lung mesenchymal cells, and periostin promoted extracellular matrix deposition, mesenchymal cell proliferation, and wound closure. Thus periostin plays a vital role in late stages of pulmonary fibrosis and is a potential biomarker for disease progression and a target for therapeutic intervention.


Assuntos
Moléculas de Adesão Celular/sangue , Fibrose Pulmonar Idiopática/metabolismo , Idoso , Animais , Anticorpos Neutralizantes/farmacologia , Biomarcadores , Moléculas de Adesão Celular/biossíntese , Proliferação de Células , Colágeno/metabolismo , Progressão da Doença , Matriz Extracelular/metabolismo , Feminino , Fibroblastos/metabolismo , Humanos , Masculino , Camundongos , Pessoa de Meia-Idade , Monócitos/metabolismo , Fator de Crescimento Transformador beta/farmacologia , Cicatrização
2.
PLoS One ; 7(2): e31336, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22363622

RESUMO

In bronchopulmonary dysplasia (BPD), alveolar septae are thickened with collagen and α-smooth muscle actin, transforming growth factor (TGF)-ß-positive myofibroblasts. Periostin, a secreted extracellular matrix protein, is involved in TGF-ß-mediated fibrosis and myofibroblast differentiation. We hypothesized that periostin expression is required for hypoalveolarization and interstitial fibrosis in hyperoxia-exposed neonatal mice, an animal model for this disease. We also examined periostin expression in neonatal lung mesenchymal stromal cells and lung tissue of hyperoxia-exposed neonatal mice and human infants with BPD. Two-to-three day-old wild-type and periostin null mice were exposed to air or 75% oxygen for 14 days. Mesenchymal stromal cells were isolated from tracheal aspirates of premature infants. Hyperoxic exposure of neonatal mice increased alveolar wall periostin expression, particularly in areas of interstitial thickening. Periostin co-localized with α-smooth muscle actin, suggesting synthesis by myofibroblasts. A similar pattern was found in lung sections of infants dying of BPD. Unlike wild-type mice, hyperoxia-exposed periostin null mice did not show larger air spaces or α-smooth muscle-positive myofibroblasts. Compared to hyperoxia-exposed wild-type mice, hyperoxia-exposed periostin null mice also showed reduced lung mRNA expression of α-smooth muscle actin, elastin, CXCL1, CXCL2 and CCL4. TGF-ß treatment increased mesenchymal stromal cell periostin expression, and periostin treatment increased TGF-ß-mediated DNA synthesis and myofibroblast differentiation. We conclude that periostin expression is increased in the lungs of hyperoxia-exposed neonatal mice and infants with BPD, and is required for hyperoxia-induced hypoalveolarization and interstitial fibrosis.


Assuntos
Moléculas de Adesão Celular/deficiência , Hiperóxia/patologia , Hiperóxia/prevenção & controle , Alvéolos Pulmonares/patologia , Idoso , Idoso de 80 Anos ou mais , Animais , Animais Recém-Nascidos , Displasia Broncopulmonar/complicações , Displasia Broncopulmonar/metabolismo , Displasia Broncopulmonar/patologia , Moléculas de Adesão Celular/genética , Moléculas de Adesão Celular/metabolismo , Diferenciação Celular/efeitos dos fármacos , DNA/biossíntese , Feminino , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Técnicas de Inativação de Genes , Humanos , Hiperóxia/complicações , Hiperóxia/genética , Hipoventilação/complicações , Hipoventilação/metabolismo , Hipoventilação/patologia , Recém-Nascido , Masculino , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Pessoa de Meia-Idade , Miofibroblastos/efeitos dos fármacos , Miofibroblastos/metabolismo , Miofibroblastos/patologia , Fenótipo , Alvéolos Pulmonares/efeitos dos fármacos , Alvéolos Pulmonares/crescimento & desenvolvimento , Alvéolos Pulmonares/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fator de Crescimento Transformador beta/farmacologia
3.
Am J Respir Cell Mol Biol ; 45(3): 445-52, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21421906

RESUMO

Prostaglandin (PG)E(2) is a bioactive eicosanoid that regulates many biologically important processes in part due to its ability to signal through four distinct G-protein-coupled receptors with differential signaling activity and unique expression patterns in different cell types. Although PGE(2) has been linked to malignancy in many organs, it is believed to play a beneficial role in the setting of fibrotic lung disease. This is in part due to the ability of PGE(2) to limit many of the pathobiologic features of lung fibroblasts and myofibroblasts, including the ability of PGE(2) to limit fibroblast proliferation, migration, collagen secretion, and, as originally reported in the Journal by us in 2003, the ability to limit transforming growth factor (TGF)-ß-induced myofibroblast differentiation. In the setting of lung fibrosis, PGE(2) production and signaling is often diminished. In the last 8 years, significant advances have been made to better understand the dysregulation of PGE(2) production and signaling in the setting of lung fibrosis. We also have a clearer picture of how PGE(2) inhibits myofibroblast differentiation and the receptor signaling pathways that can influence fibroblast proliferation. This review highlights these recent advances and offers new insights into the potential ways that PGE(2) and its downstream signals can be regulated for therapeutic benefit in a disease that has no validated treatment options.


Assuntos
Dinoprostona/metabolismo , Regulação da Expressão Gênica , Fibrose Pulmonar/patologia , Animais , Adesão Celular , Diferenciação Celular , Movimento Celular , Proliferação de Células , Epigênese Genética , Fibroblastos/citologia , Humanos , Modelos Biológicos , Estrutura Terciária de Proteína , Transdução de Sinais , Fator de Crescimento Transformador beta/metabolismo
4.
Stem Cells Dev ; 20(11): 1995-2007, 2011 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-21341990

RESUMO

We have previously isolated mesenchymal stromal cells (MSCs) from the tracheal aspirates of premature neonates with respiratory distress. Although isolation of MSCs correlates with the development of bronchopulmonary dysplasia, the physiologic role of these cells remains unclear. To address this, we further characterized the cells, focusing on the issues of gene expression, origin, and cytokine expression. Microarray comparison of early passage neonatal lung MSC gene expression to cord blood MSCs and human fetal and neonatal lung fibroblast lines demonstrated that the neonatal lung MSCs differentially expressed 971 gene probes compared with cord blood MSCs, including the transcription factors Tbx2, Tbx3, Wnt5a, FoxF1, and Gli2, each of which has been associated with lung development. Compared with lung fibroblasts, 710 gene probe transcripts were differentially expressed by the lung MSCs, including IL-6 and IL-8/CXCL8. Differential chemokine expression was confirmed by protein analysis. Further, neonatal lung MSCs exhibited a pattern of Hox gene expression distinct from cord blood MSCs but similar to human fetal lung fibroblasts, consistent with a lung origin. On the other hand, limiting dilution analysis showed that fetal lung fibroblasts form colonies at a significantly lower rate than MSCs, and fibroblasts failed to undergo differentiation along adipogenic, osteogenic, and chondrogenic lineages. In conclusion, MSCs isolated from neonatal tracheal aspirates demonstrate a pattern of lung-specific gene expression, are distinct from lung fibroblasts, and secrete pro-inflammatory cytokines.


Assuntos
Expressão Gênica , Pulmão/patologia , Células-Tronco Mesenquimais/metabolismo , Síndrome do Desconforto Respiratório do Recém-Nascido/patologia , Traqueia/patologia , Antígenos de Diferenciação , Displasia Broncopulmonar/etiologia , Displasia Broncopulmonar/patologia , Diferenciação Celular , Células Cultivadas , Citocinas/metabolismo , Feminino , Sangue Fetal , Fibroblastos/metabolismo , Perfilação da Expressão Gênica , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Humanos , Imunofenotipagem , Recém-Nascido , Recém-Nascido Prematuro , Pulmão/metabolismo , Masculino , Células-Tronco Mesenquimais/fisiologia , Fenótipo , Síndrome do Desconforto Respiratório do Recém-Nascido/complicações , Síndrome do Desconforto Respiratório do Recém-Nascido/mortalidade , Sucção
5.
Pediatrics ; 126(5): e1127-33, 2010 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-20937656

RESUMO

BACKGROUND: We have isolated mesenchymal stem cells (MSCs) from tracheal aspirates of premature infants with respiratory distress. Under the influence of transforming growth factor ß, MSCs differentiate into α-smooth-muscle actin-expressing myofibroblasts. Myofibroblasts are increased in the lungs of patients with bronchopulmonary dysplasia (BPD), a chronic lung disease of prematurely born infants. OBJECTIVE: We tested whether isolation of MSCs from tracheal aspirates of premature infants with respiratory distress during the first week of life correlates with BPD. METHODS: Eighty-four infants born at a gestational age of <33 weeks and requiring mechanical ventilation were studied. Aspirates were collected during suctioning and centrifuged. Cell pellets were resuspended in culture medium and plated. Adherent cells were grown to confluence. RESULTS: MSCs were isolated from the tracheal aspirates of 56 infants; 28 aspirate samples showed no MSCs. There was no statistical difference in gestational age or birth weight between the MSC and no-MSC groups. In the MSC group, 12 infants died and 25 developed BPD, as defined by a requirement for supplemental oxygen at 36 weeks' postmenstrual age. In the no-MSC group, 6 infants died and 1 developed BPD. Accounting for potential influences of gender, birth weight, gestational age, number of tracheal aspirate samples taken, and the duration of endotracheal intubation (up to 7 days), isolation of MSCs increased the adjusted odds ratio of BPD more than 21-fold (95% confidence interval: 1.82-265.85). CONCLUSIONS: Isolation of tracheal aspirate MSCs predicts the development of BPD, which suggests that MSCs play an important role in the pathogenesis of this disease.


Assuntos
Displasia Broncopulmonar/patologia , Recém-Nascido de muito Baixo Peso , Células-Tronco Mesenquimais/patologia , Síndrome do Desconforto Respiratório do Recém-Nascido/patologia , Células Estromais/patologia , Traqueia/patologia , Peso ao Nascer , Displasia Broncopulmonar/diagnóstico , Displasia Broncopulmonar/mortalidade , Diagnóstico Precoce , Feminino , Idade Gestacional , Humanos , Recém-Nascido , Masculino , Prognóstico , Valores de Referência , Síndrome do Desconforto Respiratório do Recém-Nascido/diagnóstico , Síndrome do Desconforto Respiratório do Recém-Nascido/mortalidade , Componente Secretório/análise , Fatores Sexuais , Células-Tronco , Sucção , Taxa de Sobrevida , Fator de Crescimento Transformador beta1/análise
6.
Respir Res ; 11: 127, 2010 Sep 21.
Artigo em Inglês | MEDLINE | ID: mdl-20858250

RESUMO

BACKGROUND: Recent studies have indicated the presence of multipotent mesenchymal stromal cells (MSCs) in human lung diseases. Excess airway smooth muscle, myofibroblasts and activated fibroblasts have each been noted in asthma, suggesting that mesenchymal progenitor cells play a role in asthma pathogenesis. We therefore sought to determine whether MSCs are present in the lungs of ovalbumin (OVA)-sensitized and challenged mice, a model of allergic airways disease. METHODS: Balb/c mice were sensitized and challenged with PBS or OVA over a 25 day period. Flow cytometry as well as colony forming and differentiation potential were used to analyze the emergence of MSCs along with gene expression studies using immunochemical analyses, quantitative polymerase chain reaction (qPCR), and gene expression beadchips. RESULTS: A CD45-negative subset of cells expressed Stro-1, Sca-1, CD73 and CD105. Selection for these markers and negative selection against CD45 yielded a population of cells capable of adipogenic, osteogenic and chondrogenic differentiation. Lungs from OVA-treated mice demonstrated a greater average colony forming unit-fibroblast (CFU-F) than control mice. Sorted cells differed from unsorted lung adherent cells, exhibiting a pattern of gene expression nearly identical to bone marrow-derived sorted cells. Finally, cells isolated from the bronchoalveolar lavage of a human asthma patient showed identical patterns of cell surface markers and differentiation potential. CONCLUSIONS: In summary, allergen sensitization and challenge is accompanied by an increase of MSCs resident in the lungs that may regulate inflammatory and fibrotic responses.


Assuntos
Imunofenotipagem , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/imunologia , Células-Tronco Multipotentes/citologia , Células-Tronco Multipotentes/imunologia , Ovalbumina/farmacologia , Animais , Contagem de Células , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/imunologia , Células Cultivadas , Imunização/métodos , Pulmão/citologia , Pulmão/efeitos dos fármacos , Pulmão/imunologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos BALB C , Células-Tronco Multipotentes/efeitos dos fármacos , Células Estromais/citologia , Células Estromais/efeitos dos fármacos , Células Estromais/imunologia
7.
Am J Physiol Lung Cell Mol Physiol ; 298(6): L735-43, 2010 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-20190033

RESUMO

We have isolated mesenchymal stem cells (MSCs) from tracheal aspirates of premature infants with respiratory distress. We examined the capacity of MSCs to differentiate into myofibroblasts, cells that participate in lung development, injury, and repair. Gene expression was measured by array, qPCR, immunoblot, and immunocytochemistry. Unstimulated MSCs expressed mRNAs encoding contractile (e.g., ACTA2, TAGLN), extracellular matrix (COL1A1 and ELN), and actin-binding (DBN1, PXN) proteins, consistent with a myofibroblast phenotype, although there was little translation into immunoreactive protein. Incubation in serum-free medium increased contractile protein (ACTA2, MYH11) gene expression. MSC-conditioned medium showed substantial levels of TGF-beta1, and treatment of serum-deprived cells with a type I activin receptor-like kinase inhibitor, SB-431542, attenuated the expression of genes encoding contractile and extracellular matrix proteins. Treatment of MSCs with TGF-beta1 further induced the expression of mRNAs encoding contractile (ACTA2, MYH11, TAGLN, DES) and extracellular matrix proteins (FN1, ELN, COL1A1, COL1A2), and increased the protein expression of alpha-smooth muscle actin, myosin heavy chain, and SM22. In contrast, human bone marrow-derived MSCs failed to undergo TGF-beta1-induced myofibroblastic differentiation. Finally, primary cells from tracheal aspirates behaved in an identical manner as later passage cells. We conclude that human neonatal lung MSCs demonstrate an mRNA expression pattern characteristic of myofibroblast progenitor cells. Autocrine production of TGF-beta1 further drives myofibroblastic differentiation, suggesting that, in the absence of other signals, fibrosis represents the "default program" for neonatal lung MSC gene expression. These data are consistent with the notion that MSCs play a key role in neonatal lung injury and repair.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Pulmão/citologia , Células-Tronco Mesenquimais/metabolismo , Fator de Crescimento Transformador beta1/biossíntese , Actinas/biossíntese , Células Cultivadas , Colágeno Tipo I/biossíntese , Cadeia alfa 1 do Colágeno Tipo I , Elastina/biossíntese , Feminino , Perfilação da Expressão Gênica , Humanos , Ácidos Hidroxâmicos/farmacologia , Recém-Nascido , Recém-Nascido Prematuro , Pulmão/metabolismo , Masculino , Proteínas dos Microfilamentos/biossíntese , Proteínas Musculares/biossíntese , Neuropeptídeos/biossíntese , Paxilina/biossíntese , RNA Mensageiro/metabolismo , Síndrome do Desconforto Respiratório do Recém-Nascido/fisiopatologia , Fator de Crescimento Transformador beta1/farmacologia
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