19 results on '"Sontake V"'
Search Results
2. Dysregulated Sox9 Overexpression in Multiple Lung Cells Contributes to Severe Fibrotic Lung Disease
- Author
-
Singh, P., primary, Gajjala, P.R., additional, Ediga, H.H., additional, Vemulapalli, C.P., additional, Sontake, V., additional, Shambhu, S., additional, Sharma, M., additional, Jegga, A.G., additional, Miyazaki, H., additional, Huang, S.K., additional, Walters, M., additional, and Madala, S.K., additional
- Published
- 2023
- Full Text
- View/download PDF
3. Human Lung Stem Cell-Based Alveolospheres Provide Insights into SARS-CoV-2-Mediated Interferon Responses and Pneumocyte Dysfunction
- Author
-
Kobayashi, Y., Katsura, H., Heaton, B.E., Mikami, Y., Fritch, E.J., Edwards, C.E., Konkimalla, A., Tata, P.R., Boucher, R.C., Randell, S.H., Heaton, N.S., Lee, P.J., Baric, R.S., Tata, A., Sontake, V., and Asakura, T.
- Abstract
Tata and colleagues report defined conditions for long-term expansion and differentiation of adult human primary alveolar stem cells. Cultured AT2s are conducive to SARS-CoV-2 infection and elicit transcriptome-wide changes that mirror COVID-19 histopathology, including upregulation of inflammatory responses, cell death, and downregulation of surfactant expression, leading to pneumocyte dysfunction. �� 2020 Elsevier Inc.Coronavirus infection causes diffuse alveolar damage leading to acute respiratory distress syndrome. The absence of ex vivo models of human alveolar epithelium is hindering an understanding of coronavirus disease 2019 (COVID-19) pathogenesis. Here, we report a feeder-free, scalable, chemically defined, and modular alveolosphere culture system for the propagation and differentiation of human alveolar type 2 cells/pneumocytes derived from primary lung tissue. Cultured pneumocytes express the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) receptor angiotensin-converting enzyme receptor type-2 (ACE2) and can be infected with virus. Transcriptome and histological analysis of infected alveolospheres mirror features of COVID-19 lungs, including emergence of interferon (IFN)-mediated inflammatory responses, loss of surfactant proteins, and apoptosis. Treatment of alveolospheres with IFNs recapitulates features of virus infection, including cell death. In contrast, alveolospheres pretreated with low-dose IFNs show a reduction in viral replication, suggesting the prophylactic effectiveness of IFNs against SARS-CoV-2. Human stem cell-based alveolospheres, thus, provide novel insights into COVID-19 pathogenesis and can serve as a model for understanding human respiratory diseases.
- Published
- 2020
- Full Text
- View/download PDF
4. Pirfenidone Therapy Attenuates Lung Function Decline in a Mouse Model of TGFα-Induced Pulmonary Fibrosis
- Author
-
Sontake, V., primary, Soundararajan, D., additional, Arron, J.R., additional, Jegga, A., additional, and Madala, S.K., additional
- Published
- 2019
- Full Text
- View/download PDF
5. Aurora Kinase B Is a Positive Regulator of Proliferation and Survival of Lung-Resident Fibroblasts in Severe Fibrotic Lung Disease
- Author
-
Kasam, R.K., primary, Geereddy, B.R., additional, Sontake, V., additional, Jegga, A., additional, and Madala, S.K., additional
- Published
- 2019
- Full Text
- View/download PDF
6. SARS-CoV-2 variant of concern fitness and adaptation in primary human airway epithelia.
- Author
-
Meganck RM, Edwards CE, Mallory ML, Lee RE, Dang H, Bailey AB, Wykoff JA, Gallant SC, Zhu DR, Yount BL, Kato T, Shaffer KM, Nakano S, Cawley AM, Sontake V, Wang JR, Hagan RS, Miller MB, Tata PR, Randell SH, Tse LV, Ehre C, Okuda K, Boucher RC, and Baric RS
- Subjects
- Humans, Virus Replication, Mutation genetics, Respiratory Mucosa virology, Genetic Fitness, Animals, Epithelial Cells virology, Chlorocebus aethiops, Adaptation, Physiological genetics, Vero Cells, SARS-CoV-2 physiology, SARS-CoV-2 genetics, COVID-19 virology, COVID-19 transmission
- Abstract
The severe acute respiratory syndrome coronavirus 2 pandemic is characterized by the emergence of novel variants of concern (VOCs) that replace ancestral strains. Here, we dissect the complex selective pressures by evaluating variant fitness and adaptation in human respiratory tissues. We evaluate viral properties and host responses to reconstruct forces behind D614G through Omicron (BA.1) emergence. We observe differential replication in airway epithelia, differences in cellular tropism, and virus-induced cytotoxicity. D614G accumulates the most mutations after infection, supporting zoonosis and adaptation to the human airway. We perform head-to-head competitions and observe the highest fitness for Gamma and Delta. Under these conditions, RNA recombination favors variants encoding the B.1.617.1 lineage 3' end. Based on viral growth kinetics, Alpha, Gamma, and Delta exhibit increased fitness compared to D614G. In contrast, the global success of Omicron likely derives from increased transmission and antigenic variation. Our data provide molecular evidence to support epidemiological observations of VOC emergence., Competing Interests: Declaration of interests R.S.B. is a member of advisory boards for VaxArt, Takeda, and Invivyd; has consulted for Gilead; and has collaborative projects with Gilead, J&J, and Hillevax focused on unrelated projects., (Copyright © 2024. Published by Elsevier Inc.)
- Published
- 2024
- Full Text
- View/download PDF
7. Host range, transmissibility and antigenicity of a pangolin coronavirus.
- Author
-
Hou YJ, Chiba S, Leist SR, Meganck RM, Martinez DR, Schäfer A, Catanzaro NJ, Sontake V, West A, Edwards CE, Yount B, Lee RE, Gallant SC, Zost SJ, Powers J, Adams L, Kong EF, Mattocks M, Tata A, Randell SH, Tata PR, Halfmann P, Crowe JE Jr, Kawaoka Y, and Baric RS
- Subjects
- Cricetinae, Humans, Animals, Mice, Host Specificity, Pangolins, SARS-CoV-2 genetics, Antibodies, Viral, COVID-19 Vaccines, Mice, Inbred BALB C, COVID-19 prevention & control, Severe acute respiratory syndrome-related coronavirus
- Abstract
The pathogenic and cross-species transmission potential of SARS-CoV-2-related coronaviruses (CoVs) remain poorly characterized. Here we recovered a wild-type pangolin (Pg) CoV GD strain including derivatives encoding reporter genes using reverse genetics. In primary human cells, PgCoV replicated efficiently but with reduced fitness and showed less efficient transmission via airborne route compared with SARS-CoV-2 in hamsters. PgCoV was potently inhibited by US Food and Drug Administration approved drugs, and neutralized by COVID-19 patient sera and SARS-CoV-2 therapeutic antibodies in vitro. A pan-Sarbecovirus antibody and SARS-CoV-2 S2P recombinant protein vaccine protected BALB/c mice from PgCoV infection. In K18-hACE2 mice, PgCoV infection caused severe clinical disease, but mice were protected by a SARS-CoV-2 human antibody. Efficient PgCoV replication in primary human cells and hACE2 mice, coupled with a capacity for airborne spread, highlights an emergence potential. However, low competitive fitness, pre-immune humans and the benefit of COVID-19 countermeasures should impede its ability to spread globally in human populations., (© 2023. The Author(s).)
- Published
- 2023
- Full Text
- View/download PDF
8. Human distal lung maps and lineage hierarchies reveal a bipotent progenitor.
- Author
-
Kadur Lakshminarasimha Murthy P, Sontake V, Tata A, Kobayashi Y, Macadlo L, Okuda K, Conchola AS, Nakano S, Gregory S, Miller LA, Spence JR, Engelhardt JF, Boucher RC, Rock JR, Randell SH, and Tata PR
- Subjects
- Alveolar Epithelial Cells, Animals, Cell Differentiation, Connectome, Fibroblasts, Gene Expression Profiling, Humans, Lung Diseases, Mice, Organoids, Primates, Regeneration, Single-Cell Analysis, Cell Lineage, Lung cytology, Stem Cells cytology
- Abstract
Mapping the spatial distribution and molecular identity of constituent cells is essential for understanding tissue dynamics in health and disease. We lack a comprehensive map of human distal airways, including the terminal and respiratory bronchioles (TRBs), which are implicated in respiratory diseases
1-4 . Here, using spatial transcriptomics and single-cell profiling of microdissected distal airways, we identify molecularly distinct TRB cell types that have not-to our knowledge-been previously characterized. These include airway-associated LGR5+ fibroblasts and TRB-specific alveolar type-0 (AT0) cells and TRB secretory cells (TRB-SCs). Connectome maps and organoid-based co-cultures reveal that LGR5+ fibroblasts form a signalling hub in the airway niche. AT0 cells and TRB-SCs are conserved in primates and emerge dynamically during human lung development. Using a non-human primate model of lung injury, together with human organoids and tissue specimens, we show that alveolar type-2 cells in regenerating lungs transiently acquire an AT0 state from which they can differentiate into either alveolar type-1 cells or TRB-SCs. This differentiation programme is distinct from that identified in the mouse lung5-7 . Our study also reveals mechanisms that drive the differentiation of the bipotent AT0 cell state into normal or pathological states. In sum, our findings revise human lung cell maps and lineage trajectories, and implicate an epithelial transitional state in primate lung regeneration and disease., (© 2022. The Author(s), under exclusive licence to Springer Nature Limited.)- Published
- 2022
- Full Text
- View/download PDF
9. Effect of Sorbitol on Alpha-Crystallin Structure and Function.
- Author
-
Kumar CU, Suryavanshi U, Sontake V, Reddy PY, Sankhala RS, Swamy MJ, and Reddy GB
- Subjects
- Animals, Molecular Chaperones metabolism, Rats, Sorbitol pharmacology, Cataract, Diabetes Mellitus, Lens, Crystalline metabolism, alpha-Crystallins chemistry, alpha-Crystallins metabolism, alpha-Crystallins pharmacology
- Abstract
Loss of eye lens transparency due to cataract is the leading cause of blindness all over the world. While aggregation of lens crystallins is the most common endpoint in various types of cataracts, chaperone-like activity (CLA) of α-crystallin preventing protein aggregation is considered to be important for maintaining the eye lens transparency. Osmotic stress due to increased accumulation of sorbitol under hyperglycemic conditions is believed to be one of the mechanisms for diabetic cataract. In addition, compromised CLA of α-crystallin in diabetic cataract has been reported. However, the effect of sorbitol on the structure and function of α-crystallin has not been elucidated yet. Hence, in the present exploratory study, we described the effect of varying concentrations of sorbitol on the structure and function of α-crystallin. Alpha-crystallin purified from the rat lens was incubated with varying concentrations of sorbitol in the dark under sterile conditions for up to 5 days. At the end of incubation, structural properties and CLA were evaluated by spectroscopic methods. Interestingly, different concentrations of sorbitol showed contrasting results: at lower concentrations (5 and 50 mM) there was a decrease in CLA and subtle alterations in secondary and tertiary structure but not at higher concentrations (500 mM). Though, these results shed a light on the effect of sorbitol on α-crystallin structure-function, further studies are required to understand the mechanism of the observed effects and their implication to cataractogenesis.
- Published
- 2022
- Full Text
- View/download PDF
10. Human Lung Stem Cell-Based Alveolospheres Provide Insights into SARS-CoV-2-Mediated Interferon Responses and Pneumocyte Dysfunction.
- Author
-
Katsura H, Sontake V, Tata A, Kobayashi Y, Edwards CE, Heaton BE, Konkimalla A, Asakura T, Mikami Y, Fritch EJ, Lee PJ, Heaton NS, Boucher RC, Randell SH, Baric RS, and Tata PR
- Subjects
- Adult, Adult Stem Cells drug effects, Adult Stem Cells enzymology, Aged, Aged, 80 and over, Alveolar Epithelial Cells enzymology, Alveolar Epithelial Cells metabolism, Angiotensin-Converting Enzyme 2 metabolism, Animals, COVID-19 physiopathology, Cell Culture Techniques, Cell Differentiation, Female, Humans, Inflammation, Male, Mice, Receptors, Coronavirus metabolism, Transcriptome, Virus Replication, Adult Stem Cells virology, Alveolar Epithelial Cells drug effects, Interferons pharmacology, SARS-CoV-2 immunology, COVID-19 Drug Treatment
- Abstract
Coronavirus infection causes diffuse alveolar damage leading to acute respiratory distress syndrome. The absence of ex vivo models of human alveolar epithelium is hindering an understanding of coronavirus disease 2019 (COVID-19) pathogenesis. Here, we report a feeder-free, scalable, chemically defined, and modular alveolosphere culture system for the propagation and differentiation of human alveolar type 2 cells/pneumocytes derived from primary lung tissue. Cultured pneumocytes express the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) receptor angiotensin-converting enzyme receptor type-2 (ACE2) and can be infected with virus. Transcriptome and histological analysis of infected alveolospheres mirror features of COVID-19 lungs, including emergence of interferon (IFN)-mediated inflammatory responses, loss of surfactant proteins, and apoptosis. Treatment of alveolospheres with IFNs recapitulates features of virus infection, including cell death. In contrast, alveolospheres pretreated with low-dose IFNs show a reduction in viral replication, suggesting the prophylactic effectiveness of IFNs against SARS-CoV-2. Human stem cell-based alveolospheres, thus, provide novel insights into COVID-19 pathogenesis and can serve as a model for understanding human respiratory diseases., Competing Interests: Declaration of Interests A patent application (PCT/US20/53158) related to this work has been filed. H.K. and P.R.T. are listed as co-inventors on this application. P.R.T. serves as a consultant for Cellarity and Surrozen., (Copyright © 2020 Elsevier Inc. All rights reserved.)
- Published
- 2020
- Full Text
- View/download PDF
11. BALO: a "mini lung" model to study cell-cell interactions.
- Author
-
Sontake V and Tata PR
- Subjects
- Animals, Cell Communication, Mice, Organogenesis, Stem Cells, Lung, Organoids
- Abstract
Stem cell-derived organoid models have emerged as a valuable tool for studying organogenesis, cell-to-cell stromal communication and disease. In this issue, Vazquez-Armendariz et al (2020) report a murine lung stem cell-based bronchioalveolar organoid system and provide insights into the effect of co-culturing with immune and mesenchymal cells., (© 2020 The Authors.)
- Published
- 2020
- Full Text
- View/download PDF
12. SARS-CoV-2 Reverse Genetics Reveals a Variable Infection Gradient in the Respiratory Tract.
- Author
-
Hou YJ, Okuda K, Edwards CE, Martinez DR, Asakura T, Dinnon KH 3rd, Kato T, Lee RE, Yount BL, Mascenik TM, Chen G, Olivier KN, Ghio A, Tse LV, Leist SR, Gralinski LE, Schäfer A, Dang H, Gilmore R, Nakano S, Sun L, Fulcher ML, Livraghi-Butrico A, Nicely NI, Cameron M, Cameron C, Kelvin DJ, de Silva A, Margolis DM, Markmann A, Bartelt L, Zumwalt R, Martinez FJ, Salvatore SP, Borczuk A, Tata PR, Sontake V, Kimple A, Jaspers I, O'Neal WK, Randell SH, Boucher RC, and Baric RS
- Subjects
- Aged, Angiotensin-Converting Enzyme 2, Animals, Antibodies, Monoclonal immunology, Antibodies, Neutralizing immunology, Betacoronavirus immunology, Betacoronavirus pathogenicity, COVID-19, Cell Line, Cells, Cultured, Chlorocebus aethiops, Coronavirus Infections immunology, Coronavirus Infections therapy, Cystic Fibrosis pathology, DNA, Recombinant, Female, Furin metabolism, Humans, Immunization, Passive, Lung metabolism, Lung pathology, Lung virology, Male, Middle Aged, Nasal Mucosa metabolism, Nasal Mucosa pathology, Nasal Mucosa virology, Pandemics, Peptidyl-Dipeptidase A metabolism, Pneumonia, Viral immunology, Respiratory System pathology, SARS-CoV-2, Serine Endopeptidases metabolism, Vero Cells, Virulence, Virus Replication, COVID-19 Serotherapy, Betacoronavirus genetics, Coronavirus Infections pathology, Coronavirus Infections virology, Pneumonia, Viral pathology, Pneumonia, Viral virology, Respiratory System virology, Reverse Genetics methods
- Abstract
The mode of acquisition and causes for the variable clinical spectrum of coronavirus disease 2019 (COVID-19) remain unknown. We utilized a reverse genetics system to generate a GFP reporter virus to explore severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pathogenesis and a luciferase reporter virus to demonstrate sera collected from SARS and COVID-19 patients exhibited limited cross-CoV neutralization. High-sensitivity RNA in situ mapping revealed the highest angiotensin-converting enzyme 2 (ACE2) expression in the nose with decreasing expression throughout the lower respiratory tract, paralleled by a striking gradient of SARS-CoV-2 infection in proximal (high) versus distal (low) pulmonary epithelial cultures. COVID-19 autopsied lung studies identified focal disease and, congruent with culture data, SARS-CoV-2-infected ciliated and type 2 pneumocyte cells in airway and alveolar regions, respectively. These findings highlight the nasal susceptibility to SARS-CoV-2 with likely subsequent aspiration-mediated virus seeding to the lung in SARS-CoV-2 pathogenesis. These reagents provide a foundation for investigations into virus-host interactions in protective immunity, host susceptibility, and virus pathogenesis., Competing Interests: Declaration of Interests The authors declare no competing financial interests., (Copyright © 2020 Elsevier Inc. All rights reserved.)
- Published
- 2020
- Full Text
- View/download PDF
13. New therapeutics based on emerging concepts in pulmonary fibrosis.
- Author
-
Sontake V, Gajjala PR, Kasam RK, and Madala SK
- Subjects
- Animals, Cell Proliferation physiology, Disease Progression, Extracellular Matrix metabolism, Fibroblasts cytology, Humans, Idiopathic Pulmonary Fibrosis mortality, Idiopathic Pulmonary Fibrosis physiopathology, Myofibroblasts cytology, Survival, Idiopathic Pulmonary Fibrosis therapy, Lung physiopathology, Molecular Targeted Therapy
- Abstract
Introduction: Fibrosis is an irreversible pathological endpoint in many chronic diseases, including pulmonary fibrosis. Idiopathic pulmonary fibrosis (IPF) is a progressive and often fatal condition characterized by (myo)fibroblast proliferation and transformation in the lung, expansion of the extracellular matrix, and extensive remodeling of the lung parenchyma. Recent evidence indicates that IPF prevalence and mortality rates are growing in the United States and elsewhere. Despite decades of research on the pathogenic mechanisms of pulmonary fibrosis, few therapeutics have succeeded in the clinic, and they have failed to improve IPF patient survival. Areas covered: Based on a literature search and our own results, we discuss the key cellular and molecular responses that contribute to (myo)fibroblast actions and pulmonary fibrosis pathogenesis; this includes signaling pathways in various cells that aberrantly and persistently activate (myo)fibroblasts in fibrotic lesions and promote scar tissue formation in the lung. Expert opinion: Lessons learned from recent failures and successes with new therapeutics point toward approaches that can target multiple pro-fibrotic processes in IPF. Advances in preclinical modeling and single-cell genomics will also accelerate novel discoveries for effective treatment of IPF.
- Published
- 2019
- Full Text
- View/download PDF
14. Wilms' tumor 1 drives fibroproliferation and myofibroblast transformation in severe fibrotic lung disease.
- Author
-
Sontake V, Kasam RK, Sinner D, Korfhagen TR, Reddy GB, White ES, Jegga AG, and Madala SK
- Subjects
- Adult, Animals, Bleomycin toxicity, Cell Differentiation genetics, Cells, Cultured, Disease Models, Animal, Extracellular Matrix metabolism, Fibrosis, Gene Expression Regulation, Gene Knock-In Techniques, Humans, Idiopathic Pulmonary Fibrosis chemically induced, Idiopathic Pulmonary Fibrosis genetics, Lung cytology, Lung drug effects, Lung pathology, Male, Mice, Transgenic, Primary Cell Culture, Promoter Regions, Genetic genetics, Actins genetics, Idiopathic Pulmonary Fibrosis pathology, Myofibroblasts pathology, Repressor Proteins metabolism, WT1 Proteins metabolism
- Abstract
Wilms' tumor 1 (WT1) is a critical transcriptional regulator of mesothelial cells during lung development but is downregulated in postnatal stages and adult lungs. We recently showed that WT1 is upregulated in both mesothelial cells and mesenchymal cells in the pathogenesis of idiopathic pulmonary fibrosis (IPF), a fatal fibrotic lung disease. Although WT1-positive cell accumulation leading to severe fibrotic lung disease has been studied, the role of WT1 in fibroblast activation and pulmonary fibrosis remains elusive. Here, we show that WT1 functions as a positive regulator of fibroblast activation, including fibroproliferation, myofibroblast transformation, and extracellular matrix (ECM) production. Chromatin immunoprecipitation experiments indicate that WT1 binds directly to the promoter DNA sequence of α-smooth muscle actin (αSMA) to induce myofibroblast transformation. In support, the genetic lineage tracing identifies WT1 as a key driver of mesothelial-to-myofibroblast and fibroblast-to-myofibroblast transformation. Importantly, the partial loss of WT1 was sufficient to attenuate myofibroblast accumulation and pulmonary fibrosis in vivo. Further, our coculture studies show that WT1 upregulation leads to non-cell autonomous effects on neighboring cells. Thus, our data uncovered a pathogenic role of WT1 in IPF by promoting fibroblast activation in the peripheral areas of the lung and as a target for therapeutic intervention.
- Published
- 2018
- Full Text
- View/download PDF
15. Repetitive intradermal bleomycin injections evoke T-helper cell 2 cytokine-driven pulmonary fibrosis.
- Author
-
Singh B, Kasam RK, Sontake V, Wynn TA, and Madala SK
- Subjects
- Animals, Bleomycin administration & dosage, Cytokines immunology, Cytokines metabolism, Disease Models, Animal, Fibroblasts metabolism, Lung metabolism, Mice, Inbred BALB C, Mice, Inbred C57BL, Pulmonary Fibrosis pathology, Th2 Cells immunology, Bleomycin pharmacology, Pulmonary Fibrosis drug therapy, Pulmonary Fibrosis metabolism, Th2 Cells drug effects
- Abstract
IL-4 and IL-13 are major T-helper cell (Th) 2 cytokines implicated in the pathogenesis of several lung diseases, including pulmonary fibrosis. In this study, using a novel repetitive intradermal bleomycin model in which mice develop extensive lung fibrosis and a progressive decline in lung function compared with saline-treated control mice, we investigated profibrotic functions of Th2 cytokines. To determine the role of IL-13 signaling in the pathogenesis of bleomycin-induced pulmonary fibrosis, wild-type, IL-13, and IL-4Rα-deficient mice were treated with bleomycin, and lungs were assessed for changes in lung function and pulmonary fibrosis. Histological staining and lung function measurements demonstrated that collagen deposition and lung function decline were attenuated in mice deficient in either IL-13 or IL-4Rα-driven signaling compared with wild-type mice treated with bleomycin. Furthermore, our results demonstrated that IL-13 and IL-4Rα-driven signaling are involved in excessive migration of macrophages and fibroblasts. Notably, our findings demonstrated that IL-13-driven migration involves increased phospho-focal adhesion kinase signaling and F-actin polymerization. Importantly, in vivo findings demonstrated that IL-13 augments matrix metalloproteinase (MMP)-2 and MMP9 activity that has also been shown to increase migration and invasiveness of fibroblasts in the lungs during bleomycin-induced pulmonary fibrosis. Together, our findings demonstrate a pathogenic role for Th2-cytokine signaling that includes excessive migration and protease activity involved in severe fibrotic lung disease.
- Published
- 2017
- Full Text
- View/download PDF
16. Hsp90 regulation of fibroblast activation in pulmonary fibrosis.
- Author
-
Sontake V, Wang Y, Kasam RK, Sinner D, Reddy GB, Naren AP, McCormack FX, White ES, Jegga AG, and Madala SK
- Subjects
- Animals, Benzoquinones pharmacology, Cell Movement drug effects, Cell Proliferation, Disease Models, Animal, Extracellular Matrix metabolism, Fibroblasts drug effects, Fibroblasts pathology, Fibrosis, Gene Knockdown Techniques, HSP90 Heat-Shock Proteins antagonists & inhibitors, HSP90 Heat-Shock Proteins genetics, Humans, Idiopathic Pulmonary Fibrosis pathology, Lactams, Macrocyclic pharmacology, Lung drug effects, Lung pathology, Mice, Mice, Transgenic, Myofibroblasts, RNA, Small Interfering, Transcriptome, Transforming Growth Factor beta, Fibroblasts metabolism, HSP90 Heat-Shock Proteins metabolism, Idiopathic Pulmonary Fibrosis metabolism, Lung metabolism
- Abstract
Idiopathic pulmonary fibrosis (IPF) is a severe fibrotic lung disease associated with fibroblast activation that includes excessive proliferation, tissue invasiveness, myofibroblast transformation, and extracellular matrix (ECM) production. To identify inhibitors that can attenuate fibroblast activation, we queried IPF gene signatures against a library of small-molecule-induced gene-expression profiles and identified Hsp90 inhibitors as potential therapeutic agents that can suppress fibroblast activation in IPF. Although Hsp90 is a molecular chaperone that regulates multiple processes involved in fibroblast activation, it has not been previously proposed as a molecular target in IPF. Here, we found elevated Hsp90 staining in lung biopsies of patients with IPF. Notably, fibroblasts isolated from fibrotic lesions showed heightened Hsp90 ATPase activity compared with normal fibroblasts. 17- N -allylamino-17-demethoxygeldanamycin (17-AAG), a small-molecule inhibitor of Hsp90 ATPase activity, attenuated fibroblast activation and also TGF-β-driven effects on fibroblast to myofibroblast transformation. The loss of the Hsp90AB, but not the Hsp90AA isoform, resulted in reduced fibroblast proliferation, myofibroblast transformation, and ECM production. Finally, in vivo therapy with 17-AAG attenuated progression of established and ongoing fibrosis in a mouse model of pulmonary fibrosis, suggesting that targeting Hsp90 represents an effective strategy for the treatment of fibrotic lung disease., Competing Interests: Conflict of interest: The authors have declared that no conflict of interest exists.
- Published
- 2017
- Full Text
- View/download PDF
17. Unique and Redundant Functions of p70 Ribosomal S6 Kinase Isoforms Regulate Mesenchymal Cell Proliferation and Migration in Pulmonary Fibrosis.
- Author
-
Madala SK, Sontake V, Edukulla R, Davidson CR, Schmidt S, and Hardie WD
- Subjects
- Animals, Bleomycin, Cell Proliferation, Collagen metabolism, Fibroblasts metabolism, Fibroblasts pathology, Humans, Isoenzymes metabolism, Ki-67 Antigen metabolism, Lung metabolism, Lung pathology, Lung physiopathology, Mice, Transgenic, Phosphorylation, Pulmonary Fibrosis physiopathology, Ribosomal Protein S6 Kinases, 70-kDa deficiency, Signal Transduction, Transforming Growth Factor alpha metabolism, Cell Movement, Mesoderm pathology, Pulmonary Fibrosis enzymology, Pulmonary Fibrosis pathology, Ribosomal Protein S6 Kinases, 70-kDa metabolism
- Abstract
The p70 ribosomal S6 kinase (p70S6K) is a downstream substrate that is phosphorylated and activated by the mammalian target of rapamycin complex and regulates multiple cellular processes associated with pulmonary fibrogenesis. Two isoforms of the p70S6K have been identified (S6K1 and S6K2), but their relative contributions in mediating pulmonary fibrosis are unknown. To interrogate the roles of the p70S6K isoforms, we overexpressed transforming growth factor (TGF)-α in mice deficient for the S6K1 or S6K2 genes and measured changes in lung histology, morphometry, total lung collagen, lung function, and proliferation between wild-type and isoform-deficient mice. Deficiency of S6K1, but not S6K2, had a significant effect on reducing proliferation in subpleural fibrotic lesions during TGF-α-induced fibrosis. Migration was significantly decreased in mesenchymal cells isolated from the lungs of S6K1 knockout mice compared with wild-type or S6K2 knockout mice. Conversely, increases in subpleural thickening were significantly decreased in S6K2-deficient mice compared with wild type. Deficiency of S6K2 significantly reduced phosphorylation of the downstream S6 ribosomal protein in lung homogenates and isolated mesenchymal cells after TGF-α expression. However, deficiency of neither isoform alone significantly altered TGF-α-induced collagen accumulation or lung function decline in vivo. Furthermore, deficiency in neither isoform prevented changes in collagen accumulation or lung compliance decline after administration of intradermal bleomycin. Together, these findings demonstrate that the p70S6K isoforms have unique and redundant functions in mediating fibrogenic processes, including proliferation, migration, and S6 phosphorylation, signifying that both isoforms must be targeted to modulate p70S6K-mediated pulmonary fibrosis.
- Published
- 2016
- Full Text
- View/download PDF
18. Fibrocytes Regulate Wilms Tumor 1-Positive Cell Accumulation in Severe Fibrotic Lung Disease.
- Author
-
Sontake V, Shanmukhappa SK, DiPasquale BA, Reddy GB, Medvedovic M, Hardie WD, White ES, and Madala SK
- Subjects
- Animals, Extracellular Matrix Proteins genetics, Extracellular Matrix Proteins immunology, Female, Humans, Idiopathic Pulmonary Fibrosis genetics, Idiopathic Pulmonary Fibrosis pathology, Lung pathology, Male, Mice, Mice, Transgenic, Repressor Proteins genetics, Transforming Growth Factor alpha genetics, Transforming Growth Factor alpha immunology, WT1 Proteins genetics, Gene Expression Regulation immunology, Idiopathic Pulmonary Fibrosis immunology, Lung immunology, Repressor Proteins immunology, WT1 Proteins immunology
- Abstract
Collagen-producing myofibroblast transdifferentiation is considered a crucial determinant in the formation of scar tissue in the lungs of patients with idiopathic pulmonary fibrosis. Multiple resident pulmonary cell types and bone marrow-derived fibrocytes have been implicated as contributors to fibrotic lesions because of the transdifferentiation potential of these cells into myofibroblasts. In this study, we assessed the expression of Wilms tumor 1 (WT1), a known marker of mesothelial cells, in various cell types in normal and fibrotic lungs. We demonstrate that WT1 is expressed by both mesothelial and mesenchymal cells in idiopathic pulmonary fibrosis lungs but has limited or no expression in normal human lungs. We also demonstrate that WT1(+) cells accumulate in fibrotic lung lesions, using two different mouse models of pulmonary fibrosis and WT1 promoter-driven fluorescent reporter mice. Reconstitution of bone marrow cells into a TGF-α transgenic mouse model demonstrated that fibrocytes do not transform into WT1(+) mesenchymal cells, but they do augment accumulation of WT1(+) cells in severe fibrotic lung disease. Importantly, the number of WT1(+) cells in fibrotic lesions was correlated with severity of lung disease as assessed by changes in lung function, histology, and hydroxyproline levels in mice. Finally, inhibition of WT1 expression was sufficient to attenuate collagen and other extracellular matrix gene production by mesenchymal cells from both murine and human fibrotic lungs. Thus, the results of this study demonstrate a novel association between fibrocyte-driven WT1(+) cell accumulation and severe fibrotic lung disease., (Copyright © 2015 by The American Association of Immunologists, Inc.)
- Published
- 2015
- Full Text
- View/download PDF
19. Th2 Cytokines Augment IL-31/IL-31RA Interactions via STAT6-dependent IL-31RA Expression.
- Author
-
Edukulla R, Singh B, Jegga AG, Sontake V, Dillon SR, and Madala SK
- Subjects
- Animals, Asthma etiology, Asthma pathology, Blotting, Western, Cells, Cultured, Chromatin Immunoprecipitation, Flow Cytometry, Fluorescent Antibody Technique, Inflammation etiology, Inflammation pathology, Interleukin-13 pharmacology, Interleukin-4 pharmacology, Interleukins genetics, Macrophages metabolism, Macrophages pathology, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, Mice, Knockout, RNA, Messenger genetics, Real-Time Polymerase Chain Reaction, Reverse Transcriptase Polymerase Chain Reaction, STAT6 Transcription Factor genetics, Schistosoma mansoni pathogenicity, Schistosomiasis mansoni complications, Schistosomiasis mansoni parasitology, Th2 Cells metabolism, Asthma metabolism, Gene Expression Regulation, Inflammation metabolism, Interleukins metabolism, Macrophages immunology, Receptors, Interleukin physiology, STAT6 Transcription Factor metabolism, Th2 Cells immunology
- Abstract
Interleukin 31 receptor α (IL-31RA) is a novel Type I cytokine receptor that pairs with oncostatin M receptor to mediate IL-31 signaling. Binding of IL-31 to its receptor results in the phosphorylation and activation of STATs, MAPK, and JNK signaling pathways. IL-31 plays a pathogenic role in tissue inflammation, particularly in allergic diseases. Recent studies demonstrate IL-31RA expression and signaling in non-hematopoietic cells, but this receptor is poorly studied in immune cells. Macrophages are key immune-effector cells that play a critical role in Th2-cytokine-mediated allergic diseases. Here, we demonstrate that Th2 cytokines IL-4 and IL-13 are capable of up-regulating IL-31RA expression on both peritoneal and bone marrow-derived macrophages from mice. Our data also demonstrate that IL-4Rα-driven IL-31RA expression is STAT6 dependent in macrophages. Notably, the inflammation-associated genes Fizz1 and serum amyloid A (SAA) are significantly up-regulated in M2 macrophages stimulated with IL-31, but not in IL-4 receptor-deficient macrophages. Furthermore, the absence of Type II IL-4 receptor signaling is sufficient to attenuate the expression of IL-31RA in vivo during allergic asthma induced by soluble egg antigen, which may suggest a role for IL-31 signaling in Th2 cytokine-driven inflammation and allergic responses. Our study reveals an important counter-regulatory role between Th2 cytokine and IL-31 signaling involved in allergic diseases., (© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.)
- Published
- 2015
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.