171 results on '"Andrew M. Tager"'
Search Results
2. Innate Immune Reconstitution in Humanized Bone Marrow-Liver-Thymus (HuBLT) Mice Governs Adaptive Cellular Immune Function and Responses to HIV-1 Infection
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Wilfredo F. Garcia-Beltran, Daniel T. Claiborne, Colby R. Maldini, Meredith Phelps, Vladimir Vrbanac, Marshall E. Karpel, Katharine L. Krupp, Karen A. Power, Christian L. Boutwell, Alejandro B. Balazs, Andrew M. Tager, Marcus Altfeld, and Todd M. Allen
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innate immunity ,T cells ,HIV-1 ,HuBLT ,BLT ,humanized mice ,Immunologic diseases. Allergy ,RC581-607 - Abstract
Humanized bone marrow-liver-thymus (HuBLT) mice are a revolutionary small-animal model that has facilitated the study of human immune function and human-restricted pathogens, including human immunodeficiency virus type 1 (HIV-1). These mice recapitulate many aspects of acute and chronic HIV-1 infection, but exhibit weak and variable T-cell responses when challenged with HIV-1, hindering our ability to confidently detect HIV-1–specific responses or vaccine effects. To identify the cause of this, we comprehensively analyzed T-cell development, diversity, and function in HuBLT mice. We found that virtually all HuBLT were well-reconstituted with T cells and had intact TCRβ sequence diversity, thymic development, and differentiation to memory and effector cells. However, there was poor CD4+ and CD8+ T-cell responsiveness to physiologic stimuli and decreased TH1 polarization that correlated with deficient reconstitution of innate immune cells, in particular monocytes. HIV-1 infection of HuBLT mice showed that mice with higher monocyte reconstitution exhibited greater CD8+ T cells responses and HIV-1 viral evolution within predicted HLA-restricted epitopes. Thus, T-cell responses to immune challenges are blunted in HuBLT mice due to a deficiency of innate immune cells, and future efforts to improve the model for HIV-1 immune response and vaccine studies need to be aimed at restoring innate immune reconstitution.
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- 2021
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3. Inhibition of CTGF ameliorates peritoneal fibrosis through suppression of fibroblast and myofibroblast accumulation and angiogenesis
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Norihiko Sakai, Miki Nakamura, Kenneth E. Lipson, Taito Miyake, Yasutaka Kamikawa, Akihiro Sagara, Yasuyuki Shinozaki, Shinji Kitajima, Tadashi Toyama, Akinori Hara, Yasunori Iwata, Miho Shimizu, Kengo Furuichi, Shuichi Kaneko, Andrew M. Tager, and Takashi Wada
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Medicine ,Science - Abstract
Abstract Peritoneal fibrosis (PF) is a serious complication in various clinical settings, but the mechanisms driving it remain to be fully determined. Connective tissue growth factor (CTGF) is known to regulate fibroblast activities. We therefore examined if CTGF inhibition has anti-fibrotic effects in PF. PF was induced by repetitive intraperitoneal injections of chlorhexidine gluconate (CG) in mice with type I pro-collagen promoter-driven green fluorescent protein (GFP) expression to identify fibroblasts. FG-3019, an anti-CTGF monoclonal antibody, was used to inhibit CTGF. CG-induced PF was significantly attenuated in FG-3019-treated mice. CG challenges induced marked accumulations of proliferating fibroblasts and of myofibroblasts, which were both reduced by FG-3019. Levels of peritoneal CTGF expression were increased by CG challenges, and suppressed in FG-3019-treated mice. FG-3019 treatment also reduced the number of CD31+ vessels and VEGF-A-positive cells in fibrotic peritoneum. In vitro studies using NIH 3T3 fibroblasts and peritoneal mesothelial cells (PMCs) showed that CTGF blockade suppressed TGF-β1-induced fibroblast proliferation and myofibroblast differentiation, PMC mesothelial-to-mesenchymal transition, and VEGF-A production. These findings suggest that the inhibition of CTGF by FG-3019 might be a novel treatment for PF through the regulation of fibroblast and myofibroblast accumulation and angiogenesis.
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- 2017
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4. HIV-1 and SIV Infection Are Associated with Early Loss of Lung Interstitial CD4+ T Cells and Dissemination of Pulmonary Tuberculosis
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Björn Corleis, Allison N. Bucsan, Maud Deruaz, Vladimir D. Vrbanac, Antonella C. Lisanti-Park, Samantha J. Gates, Alice H. Linder, Jeffrey M. Paer, Gregory S. Olson, Brittany A. Bowman, Abigail E. Schiff, Benjamin D. Medoff, Andrew M. Tager, Andrew D. Luster, Shabaana A. Khader, Deepak Kaushal, and Douglas S. Kwon
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Biology (General) ,QH301-705.5 - Abstract
Summary: Lung interstitial CD4+ T cells are critical for protection against pulmonary infections, but the fate of this population during HIV-1 infection is not well described. We studied CD4+ T cells in the setting of HIV-1 infection in human lung tissue, humanized mice, and a Mycobacterium tuberculosis (Mtb)/simian immunodeficiency virus (SIV) nonhuman primate co-infection model. Infection with a CCR5-tropic strain of HIV-1 or SIV results in severe and rapid loss of lung interstitial CD4+ T cells but not blood or lung alveolar CD4+ T cells. This is accompanied by high HIV-1 production in these cells in vitro and in vivo. Importantly, during early SIV infection, loss of lung interstitial CD4+ T cells is associated with increased dissemination of pulmonary Mtb infection. We show that lung interstitial CD4+ T cells serve as an efficient target for HIV-1 and SIV infection that leads to their early depletion and an increased risk of disseminated tuberculosis. : Corleis et al. show that lung parenchymal CD4+ T cells are permissive to HIV-1-dependent cell death. CD4+ T cell loss is highly significant in the interstitium but not the alveolar space, and loss of interstitial CD4+ T cells is associated with extrapulmonary dissemination of M. tuberculosis. Keywords: HIV-1, CD4+ T cells, cell death, tuberculosis, HIV/TB co-infection, lung, BAL
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- 2019
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5. TREX1 Knockdown Induces an Interferon Response to HIV that Delays Viral Infection in Humanized Mice
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Lee Adam Wheeler, Radiana T. Trifonova, Vladimir Vrbanac, Natasha S. Barteneva, Xing Liu, Brooke Bollman, Lauren Onofrey, Sachin Mulik, Shahin Ranjbar, Andrew D. Luster, Andrew M. Tager, and Judy Lieberman
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Biology (General) ,QH301-705.5 - Abstract
Despite their antiviral effect, the in vivo effect of interferons on HIV transmission is difficult to predict, because interferons also activate and recruit HIV-susceptible cells to sites of infection. HIV does not normally induce type I interferons in infected cells, but does if TREX1 is knocked down. Here, we investigated the effect of topical TREX1 knockdown and local interferon production on HIV transmission in human cervicovaginal explants and humanized mice. In explants in which TREX1 was knocked down, HIV induced interferons, which blocked infection. In humanized mice, even though TREX1 knockdown increased infiltrating immune cells, it delayed viral replication for 3–4 weeks. Similarly intravaginal application of type I interferons the day before HIV infection induced interferon responsive genes, reduced inflammation, and decreased viral replication. However, intravenous interferon enhanced inflammation and infection. Thus, in models of human sexual transmission, a localized interferon response inhibits HIV transmission but systemic interferons do not.
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- 2016
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6. The autotaxin-lysophosphatidic acid pathway emerges as a therapeutic target to prevent liver cancer
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Derek J. Erstad, Andrew M. Tager, Yujin Hoshida, and Bryan C. Fuchs
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atx ,chemoprevention ,cirrhosis ,fibrosis ,hcc ,hepatocellular carcinoma ,lpa ,lpar1 ,nash ,non-alcoholic steatohepatitis ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Using transcriptome meta-analysis, we recently identified the autotaxin (ATX)-lysophosphatidic acid (LPA) pathway as a regulator of hepatocellular carcinoma (HCC) risk in human cirrhosis patients. Pharmacological targeting of this pathway reduced fibrosis progression and HCC development in animals, identifying ATX-LPA signaling as a novel chemoprevention strategy for cirrhosis and HCC.
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- 2017
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7. Correction: Graft versus Host Disease in the Bone Marrow, Liver and Thymus Humanized Mouse Model.
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Matthew B. Greenblatt, Vladimir Vrbanac, Trevor Tivey, Kelly Tsang, Andrew M. Tager, and Antonios O. Aliprantis
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Medicine ,Science - Published
- 2013
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8. Screening for Inhibitors of YAP Nuclear Localization Identifies Aurora Kinase A as a Modulator of Lung Fibrosis
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Yang Yang, Daniela M. Santos, Lorena Pantano, Rachel Knipe, Elizabeth Abe, Amanda Logue, Gina Pronzati, Katharine E. Black, Jillian J. Spinney, Francesca Giacona, Michael Bieler, Cedrickx Godbout, Paul Nicklin, David Wyatt, Andrew M. Tager, Peter Seither, Franziska E. Herrmann, and Benjamin D. Medoff
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Cell Nucleus ,Pulmonary and Respiratory Medicine ,Pulmonary Fibrosis ,Clinical Biochemistry ,Cell Cycle Proteins ,YAP-Signaling Proteins ,Cell Biology ,Fibroblasts ,Phosphoproteins ,Idiopathic Pulmonary Fibrosis ,Mice ,Transforming Growth Factor beta ,Animals ,Humans ,Molecular Biology ,Original Research ,Adaptor Proteins, Signal Transducing ,Aurora Kinase A - Abstract
Idiopathic pulmonary fibrosis is a progressive lung disease with limited therapeutic options that is characterized by pathological fibroblast activation and aberrant lung remodeling with scar formation. YAP (Yes-associated protein) is a transcriptional coactivator that mediates mechanical and biochemical signals controlling fibroblast activation. We previously identified HMG-CoA (3-hydroxy-3-methylglutaryl coenzyme A) reductase inhibitors (statins) as YAP inhibitors based on a high-throughput small-molecule screen in primary human lung fibroblasts. Here we report that several Aurora kinase inhibitors were also identified from the top hits of this screen. MK-5108, a highly selective inhibitor for AURKA (Aurora kinase A), induced YAP phosphorylation and cytoplasmic retention and significantly reduced profibrotic gene expression in human lung fibroblasts. The inhibitory effect on YAP nuclear translocation and profibrotic gene expression is specific to inhibition of AURKA, but not Aurora kinase B or C, and is independent of the Hippo pathway kinases LATS1 and LATS2 (Large Tumor Suppressor 1 and 2). Further characterization of the effects of MK-5108 demonstrate that it inhibits YAP nuclear localization indirectly via effects on actin polymerization and TGFβ (Transforming Growth Factor β) signaling. In addition, MK-5108 treatment reduced lung collagen deposition in the bleomycin mouse model of pulmonary fibrosis. Our results reveal a novel role for AURKA in YAP-mediated profibrotic activity in fibroblasts and highlight the potential of small-molecule screens for YAP inhibitors for identification of novel agents with antifibrotic activity.
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- 2022
9. Reply to Kalverda et al.: Endobronchial Optical Coherence Tomography: Shining New Light on Diagnosing Usual Interstitial Pneumonitis?
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Sreyankar Nandy, Rebecca A. Raphaely, Ashok Muniappan, Angela Shih, Benjamin W. Roop, Amita Sharma, Colleen M. Keyes, Thomas V. Colby, Hugh G. Auchincloss, Henning A. Gaissert, Michael Lanuti, Christopher R. Morse, Harald C. Ott, John C. Wain, Cameron D. Wright, Maria L. Garcia-Moliner, Maxwell L. Smith, Paul A. VanderLaan, Sarita R. Berigei, Mari Mino-Kenudson, Nora K. Horick, Lloyd L. Liang, Diane L. Davies, Margit V. Szabari, Peter Caravan, Benjamin D. Medoff, Andrew M. Tager, Melissa J. Suter, and Lida P. Hariri
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Pulmonary and Respiratory Medicine ,Critical Care and Intensive Care Medicine - Published
- 2022
10. Ablation of lysophosphatidic acid receptor 1 attenuates hypertrophic cardiomyopathy in a mouse model
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Anna Axelsson Raja, Hiroko Wakimoto, Daniel M. DeLaughter, Daniel Reichart, Joshua Gorham, David A. Conner, Mingyue Lun, Clemens K. Probst, Norihiko Sakai, Rachel S. Knipe, Sydney B. Montesi, Barry Shea, Leonard P. Adam, Leslie A. Leinwand, William Wan, Esther Sue Choi, Eric L. Lindberg, Giannino Patone, Michela Noseda, Norbert Hübner, Christine E. Seidman, Andrew M. Tager, J. G. Seidman, and Carolyn Y. Ho
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Disease Models, Animal ,Mice ,Multidisciplinary ,Cardiovascular and Metabolic Diseases ,Animals ,Endothelial Cells ,Hypertrophy ,Cardiomyopathy, Hypertrophic ,Receptors, Lysophosphatidic Acid ,Carrier Proteins ,Fibrosis - Abstract
Myocardial fibrosis is a key pathologic feature of hypertrophic cardiomyopathy (HCM). However, the fibrotic pathways activated by HCM-causing sarcomere protein gene mutations are poorly defined. Because lysophosphatidic acid is a mediator of fibrosis in multiple organs and diseases, we tested the role of the lysophosphatidic acid pathway in HCM. Lysphosphatidic acid receptor 1 (LPAR1), a cell surface receptor, is required for lysophosphatidic acid mediation of fibrosis. We bred HCM mice carrying a pathogenic myosin heavy-chain variant (403 +/− ) with Lpar1 -ablated mice to create mice carrying both genetic changes (403 +/− LPAR1 −/− ) and assessed development of cardiac hypertrophy and fibrosis. Compared with 403 +/− LPAR1 WT , 403 +/− LPAR1 −/− mice developed significantly less hypertrophy and fibrosis. Single-nucleus RNA sequencing of left ventricular tissue demonstrated that Lpar1 was predominantly expressed by lymphatic endothelial cells (LECs) and cardiac fibroblasts. Lpar1 ablation reduced the population of LECs, confirmed by immunofluorescence staining of the LEC markers Lyve1 and Ccl21a and, by in situ hybridization, for Reln and Ccl21a . Lpar1 ablation also altered the distribution of fibroblast cell states. FB1 and FB2 fibroblasts decreased while FB0 and FB3 fibroblasts increased. Our findings indicate that Lpar1 is expressed predominantly by LECs and fibroblasts in the heart and is required for development of hypertrophy and fibrosis in an HCM mouse model. LPAR1 antagonism, including agents in clinical trials for other fibrotic diseases, may be beneficial for HCM.
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- 2023
11. Reply to Kalverda
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Sreyankar, Nandy, Rebecca A, Raphaely, Ashok, Muniappan, Angela, Shih, Benjamin W, Roop, Amita, Sharma, Colleen M, Keyes, Thomas V, Colby, Hugh G, Auchincloss, Henning A, Gaissert, Michael, Lanuti, Christopher R, Morse, Harald C, Ott, John C, Wain, Cameron D, Wright, Maria L, Garcia-Moliner, Maxwell L, Smith, Paul A, VanderLaan, Sarita R, Berigei, Mari, Mino-Kenudson, Nora K, Horick, Lloyd L, Liang, Diane L, Davies, Margit V, Szabari, Peter, Caravan, Benjamin D, Medoff, Andrew M, Tager, Melissa J, Suter, and Lida P, Hariri
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- 2022
12. Assessing the progression of systemic sclerosis by monitoring the tissue optic axis using PS-OCT
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Margit V. Szabari, Andrew F. McCrossan, Andrew M. Tager, David Lagares, Lida P. Hariri, Melissa J. Suter, and David C. Adams
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0301 basic medicine ,Skin score ,medicine.medical_specialty ,lcsh:Medicine ,Bleomycin ,Eye ,Refraction, Ocular ,01 natural sciences ,Palpation ,Article ,010309 optics ,03 medical and health sciences ,chemistry.chemical_compound ,Mice ,Optical coherence tomography ,Fibrosis ,Region of interest ,0103 physical sciences ,Image Processing, Computer-Assisted ,Medicine ,Animals ,Humans ,In patient ,lcsh:Science ,Multidisciplinary ,Scleroderma, Systemic ,medicine.diagnostic_test ,business.industry ,lcsh:R ,Imaging and sensing ,Translational research ,medicine.disease ,Skin manifestations ,3. Good health ,Skin diseases ,Experimental models of disease ,030104 developmental biology ,chemistry ,Disease Progression ,Organ involvement ,lcsh:Q ,Radiology ,Collagen ,business ,Tomography, Optical Coherence - Abstract
The clinical assessment of fibrosis is critical to the diagnosis and management of patients with systemic sclerosis. Current clinical standards for patient assessment is to use skin fibrosis as an indicator of organ involvement, though this approach is highly subjective and relies on manual palpation. The development of a new method for accurately quantifying collagen content may therefore significantly improve the accuracy of the traditional skin score in patients with systemic sclerosis and may additionally aid in the monitoring of anti-fibrotic therapies in clinical practice. Polarization-sensitive optical coherence tomography (PS-OCT) is a high-speed volumetric imaging modality that can be used to assess birefringent tissues including collagen. In this work we demonstrate a novel computational approach using PS-OCT for the assessment of fibrosis. This approach, based on the measured distribution of optic axis values associated with a given volume of collagen orientation, characterizes fibrotic changes independently from the depth of the region of interest in the tissue. This approach has the potential to accurately quantify collagen content and orientation faster and more robustly compared to traditional PS-OCT metrics. We investigate the viability of this approach for assessing the development of fibrosis in a bleomycin induced skin fibrosis mouse model.
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- 2020
13. Specialized transendothelial dendritic cells mediate thymic T-cell selection against blood-borne macromolecules
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Kristin Rattay, Ki-Wook Kim, Elisabeth H. Vollmann, Steffen Jung, Ulrich H. von Andrian, Vladimir Vrbanac, Andrew M. Tager, Rebecca A Fuhlbrigge, Olga Barreiro, and Aude Thiriot
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Chemokine ,Stromal cell ,Central tolerance ,Science ,Antigen-presenting cells ,CX3C Chemokine Receptor 1 ,General Physics and Astronomy ,Thymus Gland ,Autoantigens ,Article ,General Biochemistry, Genetics and Molecular Biology ,Mice ,Negative selection ,Antigen ,Cell Movement ,Animals ,Humans ,Antigen-presenting cell ,CX3CL1 ,Thymic T cell selection ,Thymocytes ,Multidisciplinary ,biology ,Chemokine CX3CL1 ,Chemistry ,Endothelial Cells ,Cell Differentiation ,Dendritic Cells ,General Chemistry ,Thymus ,Cell biology ,Mice, Inbred C57BL ,Blood ,Self Tolerance ,biology.protein - Abstract
T cells undergo rigorous selection in the thymus to ensure self-tolerance and prevent autoimmunity, with this process requiring innocuous self-antigens (Ags) to be presented to thymocytes. Self-Ags are either expressed by thymic stroma cells or transported to the thymus from the periphery by migratory dendritic cells (DCs); meanwhile, small blood-borne peptides can access the thymic parenchyma by diffusing across the vascular lining. Here we describe an additional pathway of thymic Ag acquisition that enables circulating antigenic macromolecules to access both murine and human thymi. This pathway depends on a subset of thymus-resident DCs, distinct from both parenchymal and circulating migratory DCs, that are positioned in immediate proximity to thymic microvessels where they extend cellular processes across the endothelial barrier into the blood stream. Transendothelial positioning of DCs depends on DC-expressed CX3CR1 and its endothelial ligand, CX3CL1, and disrupting this chemokine pathway prevents thymic acquisition of circulating proteins and compromises negative selection of Ag-reactive thymocytes. Thus, transendothelial DCs represent a mechanism by which the thymus can actively acquire blood-borne Ags to induce and maintain central tolerance., T cells are selected in the thymus, through interaction with self-antigens, to remove autoreactive cells. Here the authors show that a specialized thymic dendritic cell subset juxtaposes to microvessels, requires CX3CR1/CX3CL1 for this positioning, and has processes extruding into the blood stream to sample soluble macromolecules and assist in T cell selection.
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- 2021
14. Endothelial-Specific Loss of Sphingosine-1-Phosphate Receptor 1 Increases Vascular Permeability and Exacerbates Bleomycin-induced Pulmonary Fibrosis
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Alicia Franklin, Katharine E. Black, Barry S. Shea, Patricia L. Brazee, Benjamin D. Medoff, Jason W. Griffith, Andreane Cartier, Francesca Giacona, Andrew Kuo, Elizabeth A Abe, Clemens K Probst, Rachel S. Knipe, Matt Drummond, Lida P. Hariri, Christina Christoffersen, Andrew M. Tager, Amanda Logue, Eric Engelbrecht, Sydney B. Montesi, Timothy Hla, and Jillian J Spinney
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Pulmonary and Respiratory Medicine ,Clinical Biochemistry ,Vascular permeability ,Mice, Transgenic ,Lung injury ,Bleomycin ,Capillary Permeability ,Idiopathic pulmonary fibrosis ,chemistry.chemical_compound ,Fibrosis ,Sphingosine ,Pulmonary fibrosis ,Medicine ,Animals ,RNA-Seq ,Molecular Biology ,Blood Coagulation ,Lung ,Sphingosine-1-Phosphate Receptors ,S1PR1 ,Original Research ,business.industry ,Endothelial Cells ,Cell Biology ,respiratory system ,medicine.disease ,Idiopathic Pulmonary Fibrosis ,respiratory tract diseases ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Phenotype ,chemistry ,Cancer research ,Lysophospholipids ,Single-Cell Analysis ,business ,Gene Deletion - Abstract
Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive disease which leads to significant morbidity and mortality from respiratory failure. The two drugs currently approved for clinical use slow the rate of decline in lung function but have not been shown to halt disease progression or reverse established fibrosis. Thus, new therapeutic targets are needed. Endothelial injury and the resultant vascular permeability are critical components in the response to tissue injury and are present in patients with IPF. However, it remains unclear how vascular permeability affects lung repair and fibrosis following injury. Lipid mediators such as sphingosine-1-phosphate (S1P) are known to regulate multiple homeostatic processes in the lung including vascular permeability. We demonstrate that endothelial cell–(EC) specific deletion of the S1P receptor 1 (S1PR1) in mice (EC-S1pr1(−/−)) results in increased lung vascular permeability at baseline. Following a low-dose intratracheal bleomycin challenge, EC-S1pr1(−/−) mice had increased and persistent vascular permeability compared with wild-type mice, which was strongly correlated with the amount and localization of resulting pulmonary fibrosis. EC-S1pr1(−/−) mice also had increased immune cell infiltration and activation of the coagulation cascade within the lung. However, increased circulating S1P ligand in ApoM-overexpressing mice was insufficient to protect against bleomycin-induced pulmonary fibrosis. Overall, these data demonstrate that endothelial cell S1PR1 controls vascular permeability in the lung, is associated with changes in immune cell infiltration and extravascular coagulation, and modulates the fibrotic response to lung injury.
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- 2021
15. Innate Immune Reconstitution in Humanized Bone Marrow-Liver-Thymus (HuBLT) Mice Governs Adaptive Cellular Immune Function and Responses to HIV-1 Infection
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Alejandro B. Balazs, Marshall Karpel, Daniel T. Claiborne, Colby R. Maldini, Andrew M. Tager, Christian L. Boutwell, Katharine Krupp, Meredith Phelps, Todd M. Allen, Karen A. Power, Vladimir Vrbanac, Wilfredo F. Garcia-Beltran, and Marcus Altfeld
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CD4-Positive T-Lymphocytes ,BLT ,Immunology ,T cells ,Priming (immunology) ,HIV Infections ,Mice, SCID ,Biology ,CD8-Positive T-Lymphocytes ,HuBLT ,Pathogenesis ,Mice ,Immune system ,Immune Reconstitution ,Mice, Inbred NOD ,medicine ,Immunology and Allergy ,Animals ,Humans ,Viremia ,innate immunity ,Original Research ,Innate immune system ,Monocyte ,RC581-607 ,Acquired immune system ,Biological Evolution ,Disease Models, Animal ,medicine.anatomical_structure ,humanized mice ,HIV-1 ,Bone marrow ,Immunologic diseases. Allergy ,CD8 - Abstract
Humanized bone marrow-liver-thymus (HuBLT) mice have facilitated the study of human immune function and human-restricted pathogens, including human immunodeficiency virus type 1 (HIV-1). These mice not only support robust HIV-1 infection, but also uniquely harbor long-term in vivo development of HLA-restricted human T cells and recapitulate many aspects of acute and chronic HIV-1 pathogenesis. However, HuBLT mice challenged with HIV-1 exhibit variable T-cell responses across individual mice, hindering the ability to confidently detect HIV-1–specific responses or vaccine-associated effects. To understand the source of this variability, we comprehensively analyzed T-cell development, diversity, and priming in HuBLT mice. We found that while T-cell diversity, thymic development, and differentiation appeared intact in HuBLT mice, there was a defect in T-cell function, polarization, and priming that correlated with poor reconstitution of innate immune cells, particularly monocytes. While virtually all HuBLT mice were well-reconstituted with T and B cells, there was variable reconstitution of monocytes that positively correlated with T-cell function as well as TH1 polarization. Monocyte reconstitution also positively correlated with CD8+ T-cell responses in vivo during acute HIV-1 infection and the selection of viral variants. Thus, despite proper T-cell reconstitution, a deficiency in innate immune reconstitution in HuBLT mice hinders the development of more robust antigen-specific T-cell responses, which if improved would enhance the model’s ability to better recapitulate human adaptive immunity.
- Published
- 2021
16. An injectable bone marrow–like scaffold enhances T cell immunity after hematopoietic stem cell transplantation
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Azeem Sharda, Theresa M. Raimondo, James C. Weaver, Andrew M. Tager, Ting-Yu Shih, Nisarg J. Shah, David T. Scadden, Maud Deruaz, Matthew D Kerr, David J. Mooney, Vladimir Vrbanac, and Angelo S. Mao
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0303 health sciences ,Adoptive cell transfer ,Stromal cell ,Regulatory T cell ,T cell ,medicine.medical_treatment ,Biomedical Engineering ,Bioengineering ,Hematopoietic stem cell transplantation ,Biology ,Applied Microbiology and Biotechnology ,3. Good health ,03 medical and health sciences ,surgical procedures, operative ,0302 clinical medicine ,medicine.anatomical_structure ,medicine ,Cancer research ,Molecular Medicine ,Bone marrow ,Lymphopoiesis ,Progenitor cell ,030217 neurology & neurosurgery ,030304 developmental biology ,Biotechnology - Abstract
Allogeneic hematopoietic stem cell transplantation (HSCT) is a curative treatment for multiple disorders, but deficiency and dysregulation of T cells limit its utility. Here we report a biomaterial-based scaffold that mimics features of T cell lymphopoiesis in the bone marrow. The bone marrow cryogel (BMC) releases bone morphogenetic protein-2 to recruit stromal cells and presents the Notch ligand Delta-like ligand-4 to facilitate T cell lineage specification of mouse and human hematopoietic progenitor cells. BMCs subcutaneously injected in mice at the time of HSCT enhanced T cell progenitor seeding of the thymus, T cell neogenesis and diversification of the T cell receptor repertoire. Peripheral T cell reconstitution increased ~6-fold in mouse HSCT and ~2-fold in human xenogeneic HSCT. Furthermore, BMCs promoted donor CD4+ regulatory T cell generation and improved survival after allogeneic HSCT. In comparison to adoptive transfer of T cell progenitors, BMCs increased donor chimerism, T cell generation and antigen-specific T cell responses to vaccination. BMCs may provide an off-the-shelf approach for enhancing T cell regeneration and mitigating graft-versus-host disease in HSCT.
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- 2019
17. Diagnostic Accuracy of Endobronchial Optical Coherence Tomography for the Microscopic Diagnosis of Usual Interstitial Pneumonia
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Harald C. Ott, John C. Wain, Paul A. VanderLaan, Benjamin W. Roop, Cameron D. Wright, Michael Lanuti, Henning A. Gaissert, Benjamin D. Medoff, Peter Caravan, Diane L. Davies, Maxwell L. Smith, Sarita R. Berigei, Hugh Auchincloss, Christopher R. Morse, Melissa J. Suter, Rebecca A Raphaely, Mari Mino-Kenudson, Sreyankar Nandy, Amita Sharma, Lida P. Hariri, Margit V. Szabari, Lloyd L. Liang, Ashok Muniappan, Andrew M. Tager, Colleen Keyes, Nora Horick, Maria L. Garcia-Moliner, Thomas V. Colby, and Angela R. Shih
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Pulmonary and Respiratory Medicine ,medicine.medical_specialty ,Diagnostic methods ,medicine.diagnostic_test ,business.industry ,Interstitial lung disease ,Diagnostic accuracy ,Original Articles ,respiratory system ,Critical Care and Intensive Care Medicine ,medicine.disease ,Usual interstitial pneumonitis ,Fibrosis ,humanities ,respiratory tract diseases ,Idiopathic pulmonary fibrosis ,Optical coherence tomography ,Usual interstitial pneumonia ,medicine ,In vivo microscopy ,Humans ,Radiology ,business ,Lung Diseases, Interstitial ,Tomography, Optical Coherence - Abstract
Rationale: Early, accurate diagnosis of interstitial lung disease (ILD) informs prognosis and therapy, especially in idiopathic pulmonary fibrosis (IPF). Current diagnostic methods are imperfect. High-resolution computed tomography has limited resolution, and surgical lung biopsy (SLB) carries risks of morbidity and mortality. Endobronchial optical coherence tomography (EB-OCT) is a low-risk, bronchoscope-compatible modality that images large lung volumes in vivo with microscopic resolution, including subpleural lung, and has the potential to improve the diagnostic accuracy of bronchoscopy for ILD diagnosis. Objectives: We performed a prospective diagnostic accuracy study of EB-OCT in patients with ILD with a low-confidence diagnosis undergoing SLB. The primary endpoints were EB-OCT sensitivity/specificity for diagnosis of the histopathologic pattern of usual interstitial pneumonia (UIP) and clinical IPF. The secondary endpoint was agreement between EB-OCT and SLB for diagnosis of the ILD fibrosis pattern. Methods: EB-OCT was performed immediately before SLB. The resulting EB-OCT images and histopathology were interpreted by blinded, independent pathologists. Clinical diagnosis was obtained from the treating pulmonologists after SLB, blinded to EB-OCT. Measurements and Main Results: We enrolled 31 patients, and 4 were excluded because of inconclusive histopathology or lack of EB-OCT data. Twenty-seven patients were included in the analysis (16 men, average age: 65.0 yr): 12 were diagnosed with UIP and 15 with non-UIP ILD. Average FVC and Dl(CO) were 75.3% (SD, 18.5) and 53.5% (SD, 16.4), respectively. Sensitivity and specificity of EB-OCT was 100% (95% confidence interval, 75.8–100.0%) and 100% (79.6–100%), respectively, for both histopathologic UIP and clinical diagnosis of IPF. There was high agreement between EB-OCT and histopathology for diagnosis of ILD fibrosis pattern (weighted κ: 0.87 [0.72–1.0]). Conclusions: EB-OCT is a safe, accurate method for microscopic ILD diagnosis, as a complement to high-resolution computed tomography and an alternative to SLB.
- Published
- 2021
18. Targeting acid ceramidase inhibits YAP/TAZ signaling to reduce fibrosis in mice
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Aras N. Mattis, Valerie M. Weaver, Lan Wei, Steve S. Ho, Yusuf A. Hannun, Lorena Pantano, Meghan S. Mooring, Bryan C. Fuchs, Jennifer Y. Chen, Alan C. Mullen, Regina Español-Suñer, Diego dos Santos Ferreira, Dean Yimlamai, Andrew M. Tager, Daniel Canals, Raymond T. Chung, Joe M. Segal, Vikram R. Rao, Amy Yu, Johanna R. Schaub, Mai Sedki, Caroline C. Duwaerts, Sarani Ghoshal, Hsiao-Yen Ma, Nadia M. E. Ayad, Scott M. Turner, Jimmy Kuang-Hsien Hu, Benjamin Newcomb, Megan M. Marlow, Stephanie A. Christenson, Izolda Mileva, Dean Sheppard, and Sarah Alsamman
- Subjects
0301 basic medicine ,Ceramide ,Acid Ceramidase ,Medical and Health Sciences ,Article ,03 medical and health sciences ,chemistry.chemical_compound ,Mice ,0302 clinical medicine ,Fibrosis ,medicine ,Hepatic Stellate Cells ,2.1 Biological and endogenous factors ,Animals ,Humans ,Aetiology ,Cancer ,Adaptor Proteins, Signal Transducing ,biology ,Chemistry ,Signal Transducing ,Signal transducing adaptor protein ,Adaptor Proteins ,General Medicine ,Biological Sciences ,medicine.disease ,Sphingolipid ,Ubiquitin ligase ,030104 developmental biology ,030220 oncology & carcinogenesis ,biology.protein ,Cancer research ,Hepatic stellate cell ,Hepatic fibrosis ,Signal Transduction - Abstract
Hepatic stellate cells (HSCs) drive hepatic fibrosis. Therapies that inactivate HSCs have clinical potential as antifibrotic agents. We previously identified acid ceramidase (aCDase) as an antifibrotic target. We showed that tricyclic antidepressants (TCAs) reduce hepatic fibrosis by inhibiting aCDase and increasing the bioactive sphingolipid, ceramide. We now demonstrate that targeting aCDase inhibits YAP/TAZ activity by potentiating its phosphorylation-mediated proteasomal degradation via the ubiquitin ligase adaptor protein, β-TrCP. In mouse models of fibrosis, pharmacologic inhibition of aCDase or genetic knockout of aCDase in HSCs reduces fibrosis, stromal stiffness, and YAP/TAZ activity. In patients with advanced fibrosis, aCDase expression in HSCs is increased. Consistently, a signature of the genes most downregulated by ceramide identifies patients with advanced fibrosis who could benefit from aCDase targeting. The findings implicate ceramide as a critical regulator of YAP/TAZ signaling and HSC activation and highlight aCDase as a therapeutic target for the treatment of fibrosis.
- Published
- 2020
19. Endobronchial Optical Coherence Tomography for In Vivo Microscopic Diagnosis of Pulmonary Fibrosis
- Author
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Colleen Keyes, Hugh Auchincloss, Mari Mino-Kenudson, Ashok Muniappan, Christopher R. Morse, Richard L. Kradin, Michael Lanuti, John C. Wain, Amita Sharma, Margit V. Szabari, Lloyd L. Liang, Sreyankar Nandy, Benjamin W. Roop, Andrew M. Tager, Eugene J. Mark, Diane L. Davies, Lida P. Hariri, Thomas V. Colby, Angela R. Shih, Harald C. Ott, Melissa J. Suter, David C. Adams, and Henning A. Gaissert
- Subjects
Pathology ,medicine.medical_specialty ,Optical coherence tomography ,medicine.diagnostic_test ,business.industry ,In vivo ,Pulmonary fibrosis ,Medicine ,business ,medicine.disease - Published
- 2020
20. Screening for YAP Inhibitors Identifies Statins as Modulators of Fibrosis
- Author
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Royale Nichols, Jillian J Spinney, Michael Bieler, Katharine E. Black, Yufei Lin, Lida P. Hariri, Clemens K. Probst, Paul Nicklin, Benjamin D. Medoff, Lorena Pantano, Paula Grasberger, Peter Seither, David Wyatt, Daniela M. Santos, Gina Pronzati, and Andrew M. Tager
- Subjects
Pulmonary and Respiratory Medicine ,Cytoplasm ,Simvastatin ,Pulmonary Fibrosis ,Clinical Biochemistry ,Mevalonic Acid ,Cell Cycle Proteins ,Bleomycin ,Small Molecule Libraries ,Idiopathic pulmonary fibrosis ,chemistry.chemical_compound ,Mice ,Fibrosis ,Pulmonary fibrosis ,medicine ,Animals ,Humans ,Fibroblast ,Molecular Biology ,Original Research ,Adaptor Proteins, Signal Transducing ,Cell Nucleus ,Hippo signaling pathway ,biology ,business.industry ,YAP-Signaling Proteins ,Cell Biology ,Fibroblasts ,medicine.disease ,Phosphoproteins ,medicine.anatomical_structure ,chemistry ,HMG-CoA reductase ,biology.protein ,Cancer research ,Mevalonate pathway ,Acyl Coenzyme A ,Hydroxymethylglutaryl-CoA Reductase Inhibitors ,business ,Biomarkers ,Signal Transduction - Abstract
Idiopathic pulmonary fibrosis is a lung disease with limited therapeutic options that is characterized by pathological fibroblast activation and aberrant lung remodeling with scar formation. YAP (Yes-associated protein) is a transcriptional coactivator that mediates mechanical and biochemical signals controlling fibroblast activation. In this study, we developed a high-throughput small-molecule screen for YAP inhibitors in primary human lung fibroblasts. Multiple HMG-CoA (hydroxymethylglutaryl-coenzyme A) reductase inhibitors (statins) were found to inhibit YAP nuclear localization via induction of YAP phosphorylation, cytoplasmic retention, and degradation. We further show that the mevalonate pathway regulates YAP activation, and that simvastatin treatment reduces fibrosis markers in activated human lung fibroblasts and in the bleomycin mouse model of pulmonary fibrosis. Finally, we show that simvastatin modulates YAP in vivo in mouse lung fibroblasts. Our results highlight the potential of small-molecule screens for YAP inhibitors and provide a mechanism for the antifibrotic activity of statins in idiopathic pulmonary fibrosis.
- Published
- 2020
21. In vivo diagnosis of idiopathic pulmonary fibrosis (IPF) using endobronchial OCT (Conference Presentation)
- Author
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Michael Lanuti, Sreyankar Nandy, Melissa J. Suter, Angela R. Shih, David C. Adams, Andrew M. Tager, Christopher R. Morse, Eugene J. Mark, Richard L. Kradin, Thomas V. Colby, Ashok Muniappan, Colleen Keyes, Mari Mino-Kenudson, Lida P. Hariri, Benjamin W. Roop, Amita Sharma, Margit V. Szabari, Lloyd L. Liang, Diane L. Davies, and John C. Wain
- Subjects
Pathology ,medicine.medical_specialty ,genetic structures ,business.industry ,Interstitial lung disease ,respiratory system ,Optical Biopsy ,medicine.disease ,humanities ,respiratory tract diseases ,Idiopathic pulmonary fibrosis ,In vivo ,Pulmonary fibrosis ,medicine ,In vivo microscopy ,Presentation (obstetrics) ,business - Abstract
Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive form of interstitial lung disease (ILD). High-resolution CT (HRCT) is currently used to identify macroscopic IPF features. However, for 50% of patients, ultimate diagnosis relies on microscopic features from invasive, high-risk surgical biopsies (SLBX). We conducted a pilot study to assess endobronchial OCT (EB-OCT) for in vivo diagnosis of IPF, enrolling ILD patients with nondiagnostic HRCT undergoing SLBX. EB-OCT visualized salient IPF features, including microscopic features not visible on HRCT, and accurately diagnosed IPF and non-IPF ILD as compared with SLBX. EB-OCT has the potential for in vivo microscopic diagnosis of IPF.
- Published
- 2020
22. Glycerol-3-phosphate is an FGF23 regulator derived from the injured kidney
- Author
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Marta Christov, Petra Simic, Andrew M. Tager, David B. Sykes, Sushrut S. Waikar, Clary B. Clish, Wenhan Chang, Robert E. Gerszten, Najihah Aziz, Jerold Chun, Eugene P. Rhee, Ian H. de Boer, Bryan Kestenbaum, Wondong Kim, David E. Leaf, Ravi Thadhani, Debby Ngo, Zhiqiang Cheng, Paola Divieti Pajevic, Harald Jüppner, Kerry A. Pierce, Nicolas Govea, Sammy Elmariah, Wen Zhou, and Marc N. Wein
- Subjects
0301 basic medicine ,Nephrology ,Fibroblast growth factor 23 ,Male ,Glycerol ,Kidney Disease ,urologic and male genital diseases ,Kidney ,Medical and Health Sciences ,chemistry.chemical_compound ,Mice ,0302 clinical medicine ,Bone disease ,Chronic kidney disease ,Receptors ,Lysophosphatidic acid ,Homeostasis ,2.1 Biological and endogenous factors ,Aetiology ,Receptors, Lysophosphatidic Acid ,Mice, Knockout ,Chemistry ,Acute kidney injury ,General Medicine ,Acute Kidney Injury ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Glycerophosphates ,Bone Biology ,Female ,Research Article ,medicine.medical_specialty ,Knockout ,Immunology ,Renal and urogenital ,Lysophosphatidic Acid ,Cell Line ,Phosphates ,03 medical and health sciences ,Internal medicine ,medicine ,Metabolomics ,Animals ,Humans ,LPAR1 ,medicine.disease ,Fibroblast Growth Factors ,stomatognathic diseases ,Fibroblast Growth Factor-23 ,030104 developmental biology ,Endocrinology ,Renal phosphate excretion - Abstract
Fibroblast growth factor 23 (FGF23) is a bone-derived hormone that controls blood phosphate levels by increasing renal phosphate excretion and reducing 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D] production. Disorders of FGF23 homeostasis are associated with significant morbidity and mortality, but a fundamental understanding of what regulates FGF23 production is lacking. Because the kidney is the major end organ of FGF23 action, we hypothesized that it releases a factor that regulates FGF23 synthesis. Using aptamer-based proteomics and liquid chromatography–mass spectrometry–based (LC-MS–based) metabolomics, we profiled more than 1600 molecules in renal venous plasma obtained from human subjects. Renal vein glycerol-3-phosphate (G-3-P) had the strongest correlation with circulating FGF23. In mice, exogenous G-3-P stimulated bone and bone marrow FGF23 production through local G-3-P acyltransferase–mediated (GPAT-mediated) lysophosphatidic acid (LPA) synthesis. Further, the stimulatory effect of G-3-P and LPA on FGF23 required LPA receptor 1 (LPAR1). Acute kidney injury (AKI), which increases FGF23 levels, rapidly increased circulating G-3-P in humans and mice, and the effect of AKI on FGF23 was abrogated by GPAT inhibition or Lpar1 deletion. Together, our findings establish a role for kidney-derived G-3-P in mineral metabolism and outline potential targets to modulate FGF23 production during kidney injury.
- Published
- 2020
23. The Rho Kinase Isoforms ROCK1 and ROCK2 Each Contribute to the Development of Experimental Pulmonary Fibrosis
- Author
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Katharine E. Black, Norihiko Sakai, Jillian J Spinney, Linlin Zhang, James K. Liao, Andrew M. Tager, David Lagares, Alicia Franklin, Benjamin D. Medoff, Rachel S. Knipe, Paula Grasberger, Patricia L. Brazee, Barry S. Shea, and Clemens K. Probst
- Subjects
0301 basic medicine ,Pulmonary and Respiratory Medicine ,Chemistry ,Clinical Biochemistry ,Cell Biology ,Lung injury ,medicine.disease ,Actin cytoskeleton ,03 medical and health sciences ,030104 developmental biology ,Fibrosis ,Pulmonary fibrosis ,medicine ,Cancer research ,ROCK1 ,ROCK2 ,Molecular Biology ,Rho-associated protein kinase ,Myofibroblast - Abstract
Pulmonary fibrosis is thought to result from dysregulated wound repair after repetitive lung injury. Many cellular responses to injury involve rearrangements of the actin cytoskeleton mediated by the two isoforms of the Rho-associated coiled-coil-forming protein kinase (ROCK), ROCK1 and ROCK2. In addition, profibrotic mediators such as transforming growth factor-β, thrombin, and lysophosphatidic acid act through receptors that activate ROCK. Inhibition of ROCK activation may be a potent therapeutic strategy for human pulmonary fibrosis. Pharmacological inhibition of ROCK using nonselective ROCK inhibitors has been shown to prevent fibrosis in animal models; however, the specific roles of each ROCK isoform are poorly understood. Furthermore, the pleiotropic effects of this kinase have raised concerns about on-target adverse effects of ROCK inhibition such as hypotension. Selective inhibition of one isoform might be a better-tolerated strategy. In the present study, we used a genetic approach to determine the roles of ROCK1 and ROCK2 in a mouse model of bleomycin-induced pulmonary fibrosis. Using ROCK1- or ROCK2-haploinsufficient mice, we found that reduced expression of either ROCK1 or ROCK2 was sufficient to protect them from bleomycin-induced pulmonary fibrosis. In addition, we found that both isoforms contribute to the profibrotic responses of epithelial cells, endothelial cells, and fibroblasts. Interestingly, ROCK1- and ROCK2-haploinsufficient mice exhibited similar protection from bleomycin-induced vascular leak, myofibroblast differentiation, and fibrosis; however, ROCK1-haploinsufficient mice demonstrated greater attenuation of epithelial cell apoptosis. These findings suggest that selective inhibition of either ROCK isoform has the potential to be an effective therapeutic strategy for pulmonary fibrosis.
- Published
- 2018
24. Endobronchial Optical Coherence Tomography for Low-Risk Microscopic Assessment and Diagnosis of Idiopathic Pulmonary Fibrosis In Vivo
- Author
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Richard L. Kradin, Andrew M. Tager, Eugene J. Mark, Amita Sharma, John C. Wain, Melissa J. Suter, Thomas V. Colby, David C. Adams, Lida P. Hariri, Hannah Goulart, Ashok Muniappan, Mari Mino-Kenudson, and Michael Lanuti
- Subjects
Pulmonary and Respiratory Medicine ,medicine.medical_specialty ,medicine.diagnostic_test ,Extramural ,business.industry ,030204 cardiovascular system & hematology ,Critical Care and Intensive Care Medicine ,medicine.disease ,03 medical and health sciences ,Idiopathic pulmonary fibrosis ,0302 clinical medicine ,030228 respiratory system ,Optical coherence tomography ,In vivo ,medicine ,Radiology ,Tomography ,business - Published
- 2018
25. Author Correction: An injectable bone marrow–like scaffold enhances T cell immunity after hematopoietic stem cell transplantation
- Author
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David T. Scadden, Matthew D Kerr, Theresa M. Raimondo, Vladimir Vrbanac, Angelo S. Mao, James C. Weaver, Azeem Sharda, Nisarg J. Shah, David J. Mooney, Maud Deruaz, Ting-Yu Shih, and Andrew M. Tager
- Subjects
Scaffold ,business.industry ,medicine.medical_treatment ,Biomedical Engineering ,Bioengineering ,Hematopoietic stem cell transplantation ,Applied Microbiology and Biotechnology ,Injectable bone ,Cancer research ,T cell immunity ,Molecular Medicine ,Medicine ,business ,Biotechnology - Published
- 2021
26. ADAM10-mediated ephrin-B2 shedding promotes myofibroblast activation and organ fibrosis
- Author
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Rajiv Gandhi, Andrew M. Tager, Moisés Selman, Barry S. Shea, Neil Ahluwalia, Alicia Franklin, Daniela M. Santos, Mohit Kapoor, Rachel S. Knipe, Caroline Tremblay, Katharine E. Black, Jean-Pierre Pelletier, Meryem Blati, Annie Pardo, Alba Santos, Clemens K. Probst, Hassan Fahmi, David Lagares, Jiangping Wu, Sydney B. Montesi, Murray Baron, Parisa Ghassemi-Kakroodi, Paula Grasberger, Brian Wu, and Johanne Martel-Pelletier
- Subjects
0301 basic medicine ,ADAM10 ,Ephrin-B2 ,Lung injury ,Skin Diseases ,Exocytosis ,General Biochemistry, Genetics and Molecular Biology ,Extracellular matrix ,ADAM10 Protein ,Mice ,03 medical and health sciences ,Idiopathic pulmonary fibrosis ,0302 clinical medicine ,Fibrosis ,Animals ,Medicine ,Myofibroblasts ,Lung ,Cells, Cultured ,Skin ,Mice, Knockout ,Fibroblast chemotaxis ,business.industry ,Membrane Proteins ,General Medicine ,Fibroblasts ,medicine.disease ,Idiopathic Pulmonary Fibrosis ,3. Good health ,Mice, Inbred C57BL ,Protein Transport ,030104 developmental biology ,030220 oncology & carcinogenesis ,Cancer research ,Amyloid Precursor Protein Secretases ,Wound healing ,business ,Myofibroblast - Abstract
Maladaptive wound healing responses to chronic tissue injury result in organ fibrosis. Fibrosis, which entails excessive extracellular matrix (ECM) deposition and tissue remodeling by activated myofibroblasts, leads to loss of proper tissue architecture and organ function; however, the molecular mediators of myofibroblast activation have yet to be fully identified. Here we identify soluble ephrin-B2 (sEphrin-B2) as a new profibrotic mediator in lung and skin fibrosis. We provide molecular, functional and translational evidence that the ectodomain of membrane-bound ephrin-B2 is shed from fibroblasts into the alveolar airspace after lung injury. Shedding of sEphrin-B2 promotes fibroblast chemotaxis and activation via EphB3 and/or EphB4 receptor signaling. We found that mice lacking ephrin-B2 in fibroblasts are protected from skin and lung fibrosis and that a disintegrin and metalloproteinase 10 (ADAM10) is the major ephrin-B2 sheddase in fibroblasts. ADAM10 expression is increased by transforming growth factor (TGF)-β1, and ADAM10-mediated sEphrin-B2 generation is required for TGF-β1-induced myofibroblast activation. Pharmacological inhibition of ADAM10 reduces sEphrin-B2 levels in bronchoalveolar lavage and prevents lung fibrosis in mice. Consistent with the mouse data, ADAM10-sEphrin-B2 signaling is upregulated in fibroblasts from human subjects with idiopathic pulmonary fibrosis. These results uncover a new molecular mechanism of tissue fibrogenesis and identify sEphrin-B2, its receptors EphB3 and EphB4 and ADAM10 as potential therapeutic targets in the treatment of fibrotic diseases.
- Published
- 2017
27. Humanized mouse models of latent HIV infection
- Author
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Andrew M. Tager and Maud Deruaz
- Subjects
0301 basic medicine ,Viral rebound ,T cell ,030106 microbiology ,Human immunodeficiency virus (HIV) ,HIV Infections ,Mice, Transgenic ,Biology ,Virus Replication ,medicine.disease_cause ,Mice ,03 medical and health sciences ,In vivo ,Virology ,medicine ,Animals ,Humans ,Viral Load ,Antiretroviral therapy ,Virus Latency ,Disease Models, Animal ,030104 developmental biology ,medicine.anatomical_structure ,Viral replication ,Humanized mouse ,Immunology ,HIV-1 ,Viral load - Abstract
Antiretroviral therapy can efficiently control HIV viral replication, resulting in low viral loads and sustained CD4+ T cell counts in HIV-infected persons. However, fast viral rebound occurs in most infected persons when therapy is interrupted. The principal component of persistent infection is a latent but replication-competent HIV reservoir. The long half-life of this reservoir is a major barrier to cure, and its elimination is the target of important research efforts. Animal models that can recapitulate this aspect of human infection are needed to examine the HIV reservoir in tissues in vivo, and to test eradication strategies. In this review, we will summarize recent studies using humanized mouse models to examine different aspects of the viral reservoir.
- Published
- 2017
28. Noninvasive Imaging of Human Immune Responses in a Human Xenograft Model of Graft-Versus-Host Disease
- Author
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Zeina El Habre, Vladimir Vrbanac, Eric Spierings, Loes Plaisier, Juanjo Cragnolini, Catharina H. M. J. Van Elssen, Hidde L. Ploegh, Kai W. Wucherpfennig, Andrew M. Tager, Mohammad Rashidian, Jana Sticht, Johanne T. Jacobsen, Christian Freund, Jessica R. Ingram, MUMC+: MA Med Staf Artsass Interne Geneeskunde (9), and RS: FHML non-thematic output
- Subjects
0301 basic medicine ,T-Lymphocytes ,POSITRON EMISSION TOMOGRAPHY ,Immunology ,Graft vs Host Disease ,medicine.disease_cause ,Major histocompatibility complex ,ImmunoPET ,Autoimmunity ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Antigen ,single domain antibodies ,Immunity ,medicine ,Animals ,Humans ,Radiology, Nuclear Medicine and imaging ,PEPTIDE ,IN-VIVO ,Mice, Knockout ,GvHD ,MAGIC BULLET ,biology ,business.industry ,Histocompatibility Antigens Class II ,CLASS-II EXPRESSION ,Single-Domain Antibodies ,medicine.disease ,CANCER ,Immunity, Innate ,MICE ,030104 developmental biology ,Graft-versus-host disease ,humanized mice ,PET ,ANTIBODY FRAGMENTS ,030220 oncology & carcinogenesis ,Positron-Emission Tomography ,Humanized mouse ,biology.protein ,Antibody ,Radiopharmaceuticals ,business ,GENERATION - Abstract
The immune system plays a crucial role in many diseases. Activation or suppression of immunity is often related to clinical outcome. Methods to explore the dynamics of immune responses are important to elucidate their role in conditions characterized by inflammation, such as infectious disease, cancer, or autoimmunity. Immuno-PET is a noninvasive method by which disease and immune cell infiltration can be explored simultaneously. Using radiolabeled antibodies or fragments derived from them, it is possible to image disease-specific antigens and immune cell subsets. Methods: We developed a method to noninvasively image human immune responses in a relevant humanized mouse model. We generated a camelid-derived single-domain antibody specific for human class II major histocompatibility complex products and used it to noninvasively image human immune cell reconstitution in nonobese diabetic severe combined immune deficiency gamma-/- mice reconstituted with human fetal thymus, liver, and liver-derived hematopoietic stem cells (BLT mice). Results: We showed imaging of infiltrating immunocytes in BLT mice that spontaneously developed a graft-versus-host-like condition, characterized by alopecia and blepharitis. In diseased animals, we showed an increased PET signal in the liver, attributable to infiltration of activated class II major histocompatibility complex(+) T cells. Conclusion: Noninvasive imaging of immune infiltration and activation could thus be of importance for diagnosis and evaluation of treatment of graft-versus-host disease and holds promise for other diseases characterized by inflammation.
- Published
- 2017
29. An Autotaxin/Lysophosphatidic Acid/Interleukin-6 Amplification Loop Drives Scleroderma Fibrosis
- Author
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Lance Goulet, Robert Lafyatis, Katharine E. Black, Leaya George, Andrew M. Tager, Gretchen Bain, Clemens K. Probst, Flavia V. Castelino, and Veronica A. Pace
- Subjects
0301 basic medicine ,Immunology ,Scleroderma ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Rheumatology ,Fibrosis ,Lysophosphatidic acid ,medicine ,Immunology and Allergy ,Interleukin 6 ,Fibroblast ,Gene knockdown ,integumentary system ,biology ,Lipid signaling ,medicine.disease ,030104 developmental biology ,medicine.anatomical_structure ,chemistry ,030220 oncology & carcinogenesis ,biology.protein ,Cancer research ,lipids (amino acids, peptides, and proteins) ,Autotaxin - Abstract
Objective We previously implicated the lipid mediator lysophosphatidic acid (LPA) as having a role in dermal fibrosis in systemic sclerosis (SSc). The aim of this study was to identify the role of the LPA-producing enzyme autotaxin (ATX), and to connect the ATX/LPA and interleukin-6 (IL-6) pathways in SSc. Methods We evaluated the effect of a novel ATX inhibitor, PAT-048, on fibrosis and IL-6 expression in the mouse model of bleomycin-induced dermal fibrosis. We used dermal fibroblasts from SSc patients and control subjects to evaluate LPA-induced expression of IL-6, and IL-6–induced expression of ATX. We next evaluated whether LPA-induced ATX expression is dependent on IL-6, and whether baseline IL-6 expression in fibroblasts from SSc patients is dependent on ATX. Finally, we compared ATX and IL-6 expression in the skin of patients with SSc and healthy control subjects. Results PAT-048 markedly attenuated bleomycin-induced dermal fibrosis when treatment was initiated before or after the development of fibrosis. LPA stimulated expression of IL-6 in human dermal fibroblasts, and IL-6 stimulated fibroblast expression of ATX, connecting the ATX/LPA and IL-6 pathways in an amplification loop. IL-6 knockdown abrogated LPA-induced ATX expression in fibroblasts, and ATX inhibition attenuated IL-6 expression in fibroblasts and the skin of bleomycin-challenged mice. Expression of both ATX and IL-6 was increased in SSc skin, and LPA-induced IL-6 levels and IL-6–induced ATX levels were increased in fibroblasts from SSc patients compared with controls. Conclusion ATX is required for the development and maintenance of dermal fibrosis in a mouse model of bleomycin-induced SSc and enables 2 major mediators of SSc fibrogenesis, LPA and IL-6, to amplify the production of each other. Our results suggest that concurrent inhibition of these 2 pathways may be an effective therapeutic strategy for dermal fibrosis in SSc.
- Published
- 2016
30. Selective YAP/TAZ Inhibition in Fibroblasts via Dopamine Receptor D1 Agonism Reverses Fibrosis
- Author
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Jeffrey A. Meridew, Kyoung Moo Choi, Rodney D. Britt, Delphine Sicard, Jessica L. Maiers, Andrew M. Tager, Vijay H. Shah, Andrew J. Haak, Daniel J. Kass, Giovanni Ligresti, Ana Espinosa, Qi Tan, Enis Kostallari, Daniel J. Tschumperlin, Xiaoyun Li, Seyed Mehdi Nouraie, Anja C. Roden, Christina M. Pabelick, Dakota L. Jones, David Lagares, Nunzia Caporarello, Xaralabos Varelas, Yingze Zhang, Y. S. Prakash, and Aja Aravamudhan
- Subjects
Liver Cirrhosis ,Male ,Pulmonary Fibrosis ,Cell Cycle Proteins ,Article ,Extracellular matrix ,03 medical and health sciences ,Idiopathic pulmonary fibrosis ,Bleomycin ,0302 clinical medicine ,Dopamine receptor D1 ,Fibrosis ,Dopamine ,Pulmonary fibrosis ,Hepatic Stellate Cells ,Medicine ,Animals ,Humans ,030212 general & internal medicine ,Receptor ,Lung ,Adaptor Proteins, Signal Transducing ,Cell Nucleus ,Aromatic L-amino acid decarboxylase ,Chemistry ,business.industry ,Regeneration (biology) ,Receptors, Dopamine D1 ,Mesenchymal stem cell ,YAP-Signaling Proteins ,General Medicine ,Lung Injury ,Fibroblasts ,medicine.disease ,Cell biology ,Extracellular Matrix ,Phenanthridines ,Mice, Inbred C57BL ,Protein Transport ,Phenotype ,030228 respiratory system ,Gene Expression Regulation ,Cancer research ,Dopa Decarboxylase ,Trans-Activators ,Female ,RNA Interference ,business ,medicine.drug - Abstract
Tissue fibrosis is characterized by uncontrolled deposition and diminished clearance of fibrous connective tissue proteins, ultimately leading to organ scarring. Yes-associated protein (YAP) and transcriptional co-activator with PDZ-binding motif (TAZ) have recently emerged as pivotal drivers of mesenchymal cell activation in human fibrosis. Therapeutic strategies inhibiting YAP and TAZ have been hindered by the critical role that these proteins play in regeneration and homeostasis in different cell types. Here we find that the Gα(s)-coupled dopamine receptor D1 (DRD1) is preferentially expressed in lung and liver mesenchymal cells relative to other resident cells of these organs. Agonism of DRD1 selectively inhibits YAP/TAZ function in mesenchymal cells and shifts their phenotype from pro-fibrotic to fibrosis-resolving, reversing in vitro extracellular matrix stiffening and in vivo tissue fibrosis in mouse models. Aromatic L-amino acid decarboxylase (DOPA decarboxylase, or DDC), the enzyme responsible for the final step in biosynthesis of dopamine, is decreased in the lungs of subjects with idiopathic pulmonary fibrosis, and its expression inversely correlates with disease severity, consistent with an endogenous protective role for dopamine signaling that is lost in pulmonary fibrosis. Together, these findings establish a pharmacologically-tractable and cell-selective approach to targeting YAP/TAZ via DRD1 that reverses fibrosis in mice.
- Published
- 2019
31. Peroxidase Sensitive Amplifiable Probe for Molecular Magnetic Resonance Imaging of Pulmonary Inflammation
- Author
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Nicholas J. Rotile, Diego dos Santos Ferreira, Jason W. Griffith, Clemens K. Probst, Peter Caravan, Alexei A. Bogdanov, Aurora Rodríguez-Rodríguez, Sergey Shuvaev, Katherine Graham-O' Regan, Chloe M. Jones, Andrew M. Tager, Rachel S. Knipe, and Eszter Boros
- Subjects
Serotonin ,Gadolinium ,chemistry.chemical_element ,Contrast Media ,Bioengineering ,Inflammation ,02 engineering and technology ,Oxidative phosphorylation ,01 natural sciences ,Article ,Mice ,Nuclear magnetic resonance ,Pulmonary fibrosis ,medicine ,Moiety ,Animals ,Instrumentation ,Fluid Flow and Transfer Processes ,chemistry.chemical_classification ,biology ,medicine.diagnostic_test ,Process Chemistry and Technology ,010401 analytical chemistry ,Magnetic resonance imaging ,Pneumonia ,021001 nanoscience & nanotechnology ,medicine.disease ,Magnetic Resonance Imaging ,0104 chemical sciences ,Enzyme ,chemistry ,Peroxidases ,biology.protein ,medicine.symptom ,0210 nano-technology ,Peroxidase - Abstract
An amplifiable magnetic resonance imaging (MRI) probe that combines the stability of the macrocyclic Gd-DOTAGA core with a peroxidase-reactive 5-hydroxytryptamide (5-HT) moiety is reported. The incubation of the complex under enzymatic oxidative conditions led to a 1.7-fold increase in r1 at 1.4 T that was attributed to an oligomerization of the probe upon oxidation. This probe, Gd-5-HT-DOTAGA, provided specific detection of lung inflammation by MRI in bleomycin-injured mice.
- Published
- 2019
32. Immunization of BLT Humanized Mice Redirects T Cell Responses to Gag and Reduces Acute HIV-1 Viremia
- Author
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Daniel T. Claiborne, Katharine Krupp, Todd M. Allen, Edward Seung, Elizabeth F. Mellors, Karen A. Power, Timothy Dudek, Vladimir Vrbanac, David M. Knipe, Christian L. Boutwell, Andrew M. Tager, Musie Ghebremichael, Abigail Bisesi, and Colby R. Maldini
- Subjects
CD4-Positive T-Lymphocytes ,T cell ,T-Lymphocytes ,Immunology ,Viremia ,HIV Infections ,Mice, Transgenic ,Biology ,CD8-Positive T-Lymphocytes ,Microbiology ,gag Gene Products, Human Immunodeficiency Virus ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Immune system ,Virology ,Vaccines and Antiviral Agents ,medicine ,Cytotoxic T cell ,Animals ,Humans ,030304 developmental biology ,0303 health sciences ,Viral Load ,medicine.disease ,Biological Evolution ,Disease Models, Animal ,medicine.anatomical_structure ,Immunization ,030220 oncology & carcinogenesis ,Insect Science ,Humanized mouse ,Acute Disease ,Host-Pathogen Interactions ,HIV-1 ,Viral load ,CD8 - Abstract
BLT (bone marrow-liver-thymus) humanized mice, which reconstitute a functional human immune system, develop prototypic human virus-specific CD8(+) T cell responses following infection with human immunodeficiency virus type 1 (HIV-1). We explored the utility of the BLT model for HIV-1 vaccine development by immunizing BLT mice against the conserved viral Gag protein, utilizing a rapid prime-boost protocol of poly(lactic-co-glycolic) acid microparticles and a replication-defective herpes simplex virus (HSV) recombinant vector. After HIV-1 challenge, the mice developed broad, proteome-wide gamma interferon-positive (IFN-γ(+)) T cell responses against HIV-1 that reached magnitudes equivalent to what is observed in HIV-1-infected individuals. The functionality of these responses was underscored by the consistent emergence of escape mutations in multiple CD8(+) T cell epitopes during the course of infection. Although prechallenge vaccine-induced responses were largely undetectable, the Gag immunization increased both the magnitude and the kinetics of anamnestic Gag-specific T cell responses following HIV-1 infection, and the magnitude of these postchallenge Gag-specific responses was inversely correlated with acute HIV-1 viremia. Indeed, Gag immunization was associated with a modest but significant 0.5-log reduction in HIV-1 viral load when analyzed across four experimental groups of BLT mice. Notably, the HSV vector induced elevated plasma concentrations of polarizing cytokines and chemotactic factors, including interleukin-12p70 (IL-12p70) and MIP-1α, which were positively correlated with the magnitude of Gag-specific responses. Overall, these results support the ability of BLT mice to recapitulate human pathogen-specific T cell responses and to respond to immunization; however, additional improvements to the model are required to develop a robust system for testing HIV-1 vaccine efficacy. IMPORTANCE Advances in the development of humanized mice have raised the possibility of a small-animal model for preclinical testing of an HIV-1 vaccine. Here, we describe the capacity of BLT humanized mice to mount broadly directed HIV-1-specific human T cell responses that are functionally active, as indicated by the rapid emergence of viral escape mutations. Although immunization of BLT mice with the conserved viral Gag protein did not result in detectable prechallenge responses, it did increase the magnitude and kinetics of postchallenge Gag-specific T cell responses, which was associated with a modest but significant reduction in acute HIV-1 viremia. Additionally, the BLT model revealed immunization-associated increases in the plasma concentrations of immunomodulatory cytokines and chemokines that correlated with more robust T cell responses. These data support the potential utility of the BLT humanized mouse for HIV-1 vaccine development but suggest that additional improvements to the model are warranted.
- Published
- 2019
33. A High-Throughput Small Molecule Screen for YAP Inhibitors Identifies Statins as Inhibitors of Fibroblast Activation and Pulmonary Fibrosis
- Author
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Jillian J Spinney, G. Pronzati, Katharine E. Black, Benjamin D. Medoff, Clemens K. Probst, Lorena Pantano, Andrew M. Tager, Daniela M. Santos, David Wyatt, Y. Lin, R. Nichols, and Paula Grasberger
- Subjects
medicine.anatomical_structure ,Chemistry ,Pulmonary fibrosis ,medicine ,Cancer research ,medicine.disease ,Fibroblast ,Throughput (business) ,Small molecule - Published
- 2019
34. Polarization Sensitive Optical Coherence Tomography for In Vivo Microscopic Detection of Pulmonary Fibrosis
- Author
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Benjamin W. Roop, Melissa J. Suter, David C. Adams, Andrew M. Tager, Richard L. Kradin, Eugene J. Mark, Colleen Keyes, John C. Wain, Lida P. Hariri, Amita Sharma, Margit V. Szabari, Lloyd L. Liang, Christopher R. Morse, Mari Mino-Kenudson, Ashok Muniappan, Michael Lanuti, Diane L. Davies, and Thomas V. Colby
- Subjects
Polarization sensitive ,Materials science ,Nuclear magnetic resonance ,Optical coherence tomography ,medicine.diagnostic_test ,In vivo ,Pulmonary fibrosis ,medicine ,medicine.disease - Published
- 2019
35. Blocking HIV-1 Infection by Chromosomal Integrative Expression of Human CD4 on the Surface of Lactobacillus acidophilus ATCC 4356
- Author
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Joseph Sodroski, Duoyi Hu, Joshua Wiggins, Dane Bowder, Andrew M. Tager, Shi Hua Xiang, Michael Mellon, Wenzhong Wei, and Vladimir Vrbanac
- Subjects
Male ,Sexual transmission ,Immunology ,HIV Infections ,Microbiology ,Cell Line ,Mice ,03 medical and health sciences ,Lactobacillus acidophilus ,In vivo ,Virology ,Lactobacillus ,Vaccines and Antiviral Agents ,Animals ,Humans ,Gene ,030304 developmental biology ,Mice, Knockout ,0303 health sciences ,biology ,030306 microbiology ,food and beverages ,biology.organism_classification ,Recombinant Proteins ,In vitro ,Insect Science ,CD4 Antigens ,Humanized mouse ,HIV-1 ,Female ,Bacteria - Abstract
Lactobacillus bacteria are potential delivery vehicles for biopharmaceutical molecules because they are well-recognized as safe microorganisms that naturally inhabit the human body. The goal of this study was to employ these lactobacilli to combat human immunodeficiency virus type 1 (HIV-1) infection and transmission. By using a chromosomal integration method, we engineered Lactobacillus acidophilus ATCC 4356 to display human CD4, the HIV-1 receptor, on the cell surface. Since human CD4 can bind to any infectious HIV-1 particles, the engineered lactobacilli can potentially capture HIV-1 of different subtypes and prevent infection. Our data demonstrate that the CD4-carrying bacteria are able to adsorb HIV-1 particles and reduce infection significantly in vitro and also block intrarectal HIV-1 infection in a humanized mouse model in preliminary tests in vivo. Our results support the potential of this approach to decrease the efficiency of HIV-1 sexual transmission. IMPORTANCE In the absence of an effective vaccine, alternative approaches to block HIV-1 infection and transmission with commensal bacteria expressing antiviral proteins are being considered. This report provides a proof-of-concept by using Lactobacillus bacteria stably expressing the HIV-1 receptor CD4 to capture and neutralize HIV-1 in vitro and in a humanized mouse model. The stable expression of antiviral proteins, such as CD4, following genomic integration of the corresponding genes into this Lactobacillus strain may contribute to the prevention of HIV-1 sexual transmission.
- Published
- 2019
36. The Fibrosis Across Organs Symposium: A Roadmap for Future Research Priorities
- Author
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Andrew M. Tager, Timothy A. McKinsey, Lynn M. Schnapp, Gregory P. Cosgrove, John D. Tosi, Stephen Crane, Kevin K. Brown, Jesse Roman, David A. Brenner, Dennis E. Doherty, Dolly Kervitsky, Tim D. Hewitson, Luca Richeldi, Bodh I. Jugdutt, Eric S. White, and Teresa Barnes
- Subjects
Research design ,medicine.medical_specialty ,Lung ,Colorado ,business.industry ,General Medicine ,Congresses as Topic ,medicine.disease ,Fibrosis ,Article ,Idiopathic pulmonary fibrosis ,medicine.anatomical_structure ,Research Design ,medicine ,Intensive care medicine ,business - Published
- 2019
37. Fibrogenic Lung Injury Induces Non–Cell-Autonomous Fibroblast Invasion
- Author
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Paula Grasberger, Moisés Selman, David Lagares, Brian Mugo, Annie Pardo, Carol Feghali-Bostwick, Neil Ahluwalia, and Andrew M. Tager
- Subjects
Lipopolysaccharides ,Male ,0301 basic medicine ,Pulmonary and Respiratory Medicine ,Pathology ,medicine.medical_specialty ,medicine.medical_treatment ,Clinical Biochemistry ,Lung injury ,Biology ,Bleomycin ,03 medical and health sciences ,Idiopathic pulmonary fibrosis ,Cell Movement ,Transforming Growth Factor beta ,Pulmonary fibrosis ,medicine ,Animals ,Humans ,Fibroblast ,Molecular Biology ,Original Research ,medicine.diagnostic_test ,Fibroblast growth factor receptor 2 ,Chemotaxis ,Growth factor ,Lung Injury ,Cell Biology ,Transforming growth factor beta ,Fibroblasts ,respiratory system ,medicine.disease ,Idiopathic Pulmonary Fibrosis ,respiratory tract diseases ,Mice, Inbred C57BL ,030104 developmental biology ,Bronchoalveolar lavage ,medicine.anatomical_structure ,Solubility ,Gene Knockdown Techniques ,Cancer research ,biology.protein ,Bronchoalveolar Lavage Fluid - Abstract
Pathologic accumulation of fibroblasts in pulmonary fibrosis appears to depend on their invasion through basement membranes and extracellular matrices. Fibroblasts from the fibrotic lungs of patients with idiopathic pulmonary fibrosis (IPF) have been demonstrated to acquire a phenotype characterized by increased cell-autonomous invasion. Here, we investigated whether fibroblast invasion is further stimulated by soluble mediators induced by lung injury. We found that bronchoalveolar lavage fluids from bleomycin-challenged mice or patients with IPF contain mediators that dramatically increase the matrix invasion of primary lung fibroblasts. Further characterization of this non–cell-autonomous fibroblast invasion suggested that the mediators driving this process are produced locally after lung injury and are preferentially produced by fibrogenic (e.g., bleomycin-induced) rather than nonfibrogenic (e.g., LPS-induced) lung injury. Comparison of invasion and migration induced by a series of fibroblast-active mediators indicated that these two forms of fibroblast movement are directed by distinct sets of stimuli. Finally, knockdown of multiple different membrane receptors, including platelet-derived growth factor receptor-β, lysophosphatidic acid 1, epidermal growth factor receptor, and fibroblast growth factor receptor 2, mitigated the non–cell-autonomous fibroblast invasion induced by bronchoalveolar lavage from bleomycin-injured mice, suggesting that multiple different mediators drive fibroblast invasion in pulmonary fibrosis. The magnitude of this mediator-driven fibroblast invasion suggests that its inhibition could be a novel therapeutic strategy for pulmonary fibrosis. Further elaboration of the molecular mechanisms that drive non–cell-autonomous fibroblast invasion consequently may provide a rich set of novel drug targets for the treatment of IPF and other fibrotic lung diseases.
- Published
- 2016
38. In vivo optical imaging for low-risk microscopic diagnosis of IPF
- Author
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Christopher R. Morse, Hannah Goulart, Melissa J. Suter, Hugh Auchincloss, John C. Wain, Hamid Pahlevaninezhad, Henning A. Gaissert, Colleen Keyes, Richard L. Kradin, Lida P. Hariri, Amita Sharma, Thomas V. Colby, Margit V. Szabari, Lloyd L. Liang, David H. Adams, Mari Mino-Kenudson, Michael Lanuti, Ashok Muniappan, Andrew M. Tager, and Eugene J. Mark
- Subjects
Pathology ,medicine.medical_specialty ,Optical imaging ,In vivo ,business.industry ,medicine ,business - Published
- 2018
39. Therapeutic Efficacy of Vectored PGT121 Gene Delivery in HIV-1-Infected Humanized Mice
- Author
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Michael R. Boyd, Marinela Kirilova, Vladimir Vrbanac, Peter Abbink, Dan H. Barouch, Po-Ting Liu, Christine A. Bricault, Lawrence J. Tartaglia, Noe B. Mercado, Andrew M. Tager, Ovini Nanayakkara, Rafael A. Larocca, Alexander Badamchi-Zadeh, and Michael S. Seaman
- Subjects
0301 basic medicine ,viruses ,Genetic Vectors ,Immunology ,HIV Infections ,adeno-associated virus ,Gene delivery ,Biology ,medicine.disease_cause ,Microbiology ,Virus ,Adenoviridae ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Transduction, Genetic ,Virology ,Vaccines and Antiviral Agents ,medicine ,Animals ,neutralizing antibodies ,Vector (molecular biology) ,Adeno-associated virus ,Mice, Knockout ,Mice, Inbred BALB C ,Genetic Therapy ,3. Good health ,030104 developmental biology ,adenoviruses ,030220 oncology & carcinogenesis ,Insect Science ,PGT121 ,HIV-1 ,biology.protein ,Female ,Antibody ,Viral load ,Antiantibody - Abstract
Broadly neutralizing antibodies (bNAbs) are being explored for HIV-1 prevention and cure strategies. However, administration of purified bNAbs poses challenges in resource-poor settings, where the HIV-1 disease burden is greatest. In vivo vector-based production of bNAbs represents an alternative strategy. We investigated adenovirus serotype 5 (Ad5) and adeno-associated virus serotype 1 (AAV1) vectors to deliver the HIV-1-specific bNAb PGT121 in wild-type and immunocompromised C57BL/6 mice as well as in HIV-1-infected bone marrow-liver-thymus (BLT) humanized mice. Ad5.PGT121 and AAV1.PGT121 produced functional antibody in vivo . Ad5.PGT121 produced PGT121 rapidly within 6 h, whereas AAV1.PGT121 produced detectable PGT121 in serum by 72 h. Serum PGT121 levels were rapidly reduced by the generation of anti-PGT121 antibodies in immunocompetent mice but were durably maintained in immunocompromised mice. In HIV-1-infected BLT humanized mice, Ad5.PGT121 resulted in a greater reduction of viral loads than did AAV1.PGT121. Ad5.PGT121 also led to more-sustained virologic control than purified PGT121 IgG. Ad5.PGT121 afforded more rapid, robust, and durable antiviral efficacy than AAV1.PGT121 and purified PGT121 IgG in HIV-1-infected humanized mice. Further evaluation of vector delivery of HIV-1 bNAbs is warranted, although approaches to prevent the generation of antiantibody responses may also be required. IMPORTANCE Broadly neutralizing antibodies (bNAbs) are being explored for HIV-1 prevention and cure strategies, but delivery of purified antibodies may prove challenging. We investigated adenovirus serotype 5 (Ad5) and adeno-associated virus serotype 1 (AAV1) vectors to deliver the HIV-1-specific bNAb PGT121. Ad5.PGT121 afforded more rapid, robust, and durable antiviral efficacy than AAV1.PGT121 and purified PGT121 IgG in HIV-1-infected humanized mice.
- Published
- 2018
40. Gadofosveset-enhanced lung magnetic resonance imaging to detect ongoing vascular leak in pulmonary fibrosis
- Author
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Hannah Goulart, Andrew M. Tager, Peter Caravan, Ravi Teja Seethamraju, Rohan Rao, Subba R. Digumarthy, Amita Sharma, Sydney B. Montesi, Lloyd L. Liang, and Barry S. Shea
- Subjects
0301 basic medicine ,Pulmonary and Respiratory Medicine ,medicine.medical_specialty ,Pulmonary Fibrosis ,Contrast Media ,Gadolinium ,Vascular leakage ,Lung pathology ,Article ,03 medical and health sciences ,0302 clinical medicine ,Pulmonary fibrosis ,Organometallic Compounds ,Medicine ,Humans ,Vascular Diseases ,Lung ,medicine.diagnostic_test ,business.industry ,Extramural ,Gadofosveset ,Magnetic resonance imaging ,respiratory system ,medicine.disease ,Magnetic Resonance Imaging ,respiratory tract diseases ,030104 developmental biology ,medicine.anatomical_structure ,030228 respiratory system ,Case-Control Studies ,Radiology ,business ,medicine.drug - Abstract
Vascular leak is increased diffusely in the lungs in pulmonary fibrosis and is not limited to radiographic areas of disease involvement http://ow.ly/Zz6Z30jb85l
- Published
- 2018
41. Targeted apoptosis of myofibroblasts with the BH3 mimetic ABT-263 reverses established fibrosis
- Author
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Jeremy Ryan, David Lagares, Joan Montero, Murray Baron, Anthony Letai, Tobias Kuehl, Patrick Bhola, Andrew M. Tager, Mohit Kapoor, Norihiko Sakai, Alba Santos, Paula Grasberger, Fei Liu, Daniel J. Tschumperlin, Clemens K. Probst, Xaralabos Varelas, and Rod A. Rahimi
- Subjects
Male ,0301 basic medicine ,Cell Survival ,bcl-X Protein ,Priming (immunology) ,Apoptosis ,Mitochondrion ,Mechanotransduction, Cellular ,03 medical and health sciences ,chemistry.chemical_compound ,Fibrosis ,Animals ,Humans ,Medicine ,Mechanotransduction ,Myofibroblasts ,Sulfonamides ,Aniline Compounds ,Scleroderma, Systemic ,Navitoclax ,business.industry ,Dermis ,General Medicine ,medicine.disease ,Biomechanical Phenomena ,Extracellular Matrix ,Mitochondria ,Mice, Inbred C57BL ,Disease Models, Animal ,030104 developmental biology ,chemistry ,Mitochondrial Membranes ,Cancer research ,biological phenomena, cell phenomena, and immunity ,business ,Wound healing ,Myofibroblast ,Signal Transduction - Abstract
Persistent myofibroblast activation distinguishes pathological fibrosis from physiological wound healing, suggesting that therapies selectively inducing myofibroblast apoptosis could prevent progression and potentially reverse established fibrosis in diseases such as scleroderma, a heterogeneous autoimmune disease characterized by multiorgan fibrosis. We demonstrate that fibroblast-to-myofibroblast differentiation driven by matrix stiffness increases the mitochondrial priming (proximity to the apoptotic threshold) of these activated cells. Mitochondria in activated myofibroblasts, but not quiescent fibroblasts, are primed by death signals such as the proapoptotic BH3-only protein BIM, which creates a requirement for tonic expression of the antiapoptotic protein BCL-XL to sequester BIM and ensure myofibroblast survival. Myofibroblasts become particularly susceptible to apoptosis induced by “BH3 mimetic” drugs inhibiting BCL-XL such as ABT-263. ABT-263 displaces BCL-XL binding to BIM, allowing BIM to activate apoptosis on stiffness-primed myofibroblasts. Therapeutic blockade of BCL-XL with ABT-263 (navitoclax) effectively treats established fibrosis in a mouse model of scleroderma dermal fibrosis by inducing myofibroblast apoptosis. Using a BH3 profiling assay to assess mitochondrial priming in dermal fibroblasts derived from patients with scleroderma, we demonstrate that the extent of apoptosis induced by BH3 mimetic drugs correlates with the extent of their mitochondrial priming, indicating that BH3 profiling could predict apoptotic responses of fibroblasts to BH3 mimetic drugs in patients with scleroderma. Together, our findings elucidate the potential efficacy of targeting myofibroblast antiapoptotic proteins with BH3 mimetic drugs in scleroderma and other fibrotic diseases.
- Published
- 2017
42. Corrigendum: ADAM10-mediated ephrin-B2 shedding promotes myofibroblast activation and organ fibrosis
- Author
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Jean-Pierre Pelletier, Parisa Ghassemi-Kakroodi, Alba Santos, Murray Baron, Mohit Kapoor, Caroline Tremblay, Rachel S. Knipe, Moisés Selman, Katharine E. Black, Hassan Fahmi, Brian Wu, Sydney B. Montesi, Neil Ahluwalia, Andrew M. Tager, Johanne Martel-Pelletier, David Lagares, Alicia Franklin, Meryem Blati, Paula Grasberger, Annie Pardo, Daniela M. Santos, Jiangping Wu, Rajiv Gandhi, Barry S. Shea, and Clemens K. Probst
- Subjects
0301 basic medicine ,animal structures ,030102 biochemistry & molecular biology ,business.industry ,ADAM10 ,macromolecular substances ,General Medicine ,medicine.disease ,General Biochemistry, Genetics and Molecular Biology ,Article ,03 medical and health sciences ,nervous system ,Fibrosis ,embryonic structures ,medicine ,Cancer research ,sense organs ,business ,Ephrin b2 ,Myofibroblast - Abstract
Maladaptive wound healing responses to chronic tissue injury result in organ fibrosis. Fibrosis, which entails excessive extracellular matrix (ECM) deposition and tissue remodelling by activated myofibroblasts, leads to loss of proper tissue architecture and organ function; however the molecular mediators of myofibroblast activation remain to be fully identified. Here we identify soluble ephrin-B2 as a novel pro-fibrotic mediator in lung and skin fibrosis. We provide molecular, functional and translational evidence that the ectodomain of membrane-bound ephrin-B2 is shed from fibroblasts into the alveolar airspace after lung injury. Shedding of soluble ephrin-B2 (sEphrin-B2) promotes fibroblast chemotaxis and activation via EphB3/EphB4 receptor signaling. We found that mice lacking ephrin-B2 in fibroblasts are protected from skin and lung fibrosis and that a distintegrin and metalloproteinase 10 (ADAM10) is the major ephrin-B2 sheddase in fibroblasts. ADAM10 is induced by transforming growth factor-β1 (TGF-β1), and ADAM10-mediated sEphrin-B2 generation is required for TGF-β1–induced myofibroblast activation. Pharmacological inhibition of ADAM10 reduces sEphrin-B2 levels in bronchoalveolar lavage and prevents lung fibrosis in mice. Consistent with the mouse data, ADAM10/sEphrin-B2 signaling is upregulated in fibroblasts from human subjects with idiopathic pulmonary fibrosis. These results uncover a new molecular mechanism of tissue fibrogenesis and identify sEphrin-B2, its receptors Eph3/Eph4, and ADAM10 as potential therapeutic targets in the treatment of fibrotic diseases.
- Published
- 2017
43. An injectable bone marrow-like scaffold enhances T cell immunity after hematopoietic stem cell transplantation
- Author
-
Nisarg J, Shah, Angelo S, Mao, Ting-Yu, Shih, Matthew D, Kerr, Azeem, Sharda, Theresa M, Raimondo, James C, Weaver, Vladimir D, Vrbanac, Maud, Deruaz, Andrew M, Tager, David J, Mooney, and David T, Scadden
- Subjects
Tissue Scaffolds ,Transplantation, Heterologous ,Hematopoietic Stem Cell Transplantation ,Graft vs Host Disease ,Hematopoietic Stem Cells ,Adoptive Transfer ,Chimerism ,T-Lymphocytes, Regulatory ,Mice ,Bone Marrow ,Animals ,Humans ,Transplantation, Homologous ,Bone Marrow Transplantation - Abstract
Allogeneic hematopoietic stem cell transplantation (HSCT) is a curative treatment for multiple disorders, but deficiency and dysregulation of T cells limit its utility. Here we report a biomaterial-based scaffold that mimics features of T cell lymphopoiesis in the bone marrow. The bone marrow cryogel (BMC) releases bone morphogenetic protein-2 to recruit stromal cells and presents the Notch ligand Delta-like ligand-4 to facilitate T cell lineage specification of mouse and human hematopoietic progenitor cells. BMCs subcutaneously injected in mice at the time of HSCT enhanced T cell progenitor seeding of the thymus, T cell neogenesis and diversification of the T cell receptor repertoire. Peripheral T cell reconstitution increased ~6-fold in mouse HSCT and ~2-fold in human xenogeneic HSCT. Furthermore, BMCs promoted donor CD4
- Published
- 2017
44. A Small-Molecule CD4-Mimetic Compound Protects Bone Marrow-Liver-Thymus Humanized Mice From HIV-1 Infection
- Author
-
Bruno Melillo, Vladimir Vrbanac, Amos B. Smith, Navid Madani, Andrew M. Tager, Amy M. Princiotto, Jongwoo Park, and Joseph Sodroski
- Subjects
0301 basic medicine ,Sexual transmission ,CD4 antigen ,030106 microbiology ,HIV Infections ,Mice, SCID ,Thymus Gland ,Virus ,03 medical and health sciences ,chemistry.chemical_compound ,Major Articles and Brief Reports ,Viral envelope ,Biomimetics ,Bone Marrow ,HIV Fusion Inhibitors ,medicine ,Immunology and Allergy ,Animals ,Receptor ,chemistry.chemical_classification ,business.industry ,Virology ,Administration, Intravaginal ,Disease Models, Animal ,030104 developmental biology ,Infectious Diseases ,medicine.anatomical_structure ,Treatment Outcome ,chemistry ,Liver ,CD4 Antigens ,Vagina ,HIV-1 ,Female ,Bone marrow ,business ,Glycoprotein - Abstract
Background Small-molecule CD4-mimetic compounds (CD4mc) inhibit human immunodeficiency virus (HIV-1) entry by blocking binding to the CD4 receptor and by premature triggering of the viral envelope glycoprotein (Env) spike. Methods The efficacy of a CD4mc in protecting bone marrow-liver-thymus (BLT) humanized mice from vaginal HIV-1 challenge was evaluated. Results Intravaginal application of the CD4mc JP-III-48, either before or simultaneously with virus challenge, protected BLT humanized mice from HIV-1JR-CSF infection in a dose- dependent manner. Conclusion The direct antiviral effects of a CD4mc prevent HIV-1 infection in a murine model of sexual transmission.
- Published
- 2017
45. Prevention vaginally of HIV-1 transmission in humanized BLT mice and mode of antiviral action of polyanionic carbosilane dendrimer G2-S16
- Author
-
María Jesús Serramía, Francisco Javier de la Mata, Rafael Gómez, Daniel Sepúlveda-Crespo, Andrew M. Tager, Ma Ángeles Muñoz-Fernández, Jose L. Jimenez, and Vladimir Vrbanac
- Subjects
Alkanesulfonates ,Dendrimers ,Sexual transmission ,Anti-HIV Agents ,Biomedical Engineering ,Pharmaceutical Science ,Medicine (miscellaneous) ,HIV Infections ,Bioengineering ,Virus ,Mice ,Microbicide ,Dendrimer ,Animals ,Humans ,Medicine ,Organosilicon Compounds ,General Materials Science ,business.industry ,Transmission (medicine) ,Vaginal microbicide ,Virology ,Hiv 1 transmission ,Viral replication ,Vagina ,HIV-1 ,Molecular Medicine ,Female ,business - Abstract
The development of a safe, effective, and low-priced topical microbicide to prevent HIV-1 sexual transmission is urgently needed. The emerging field of nanotechnology plays an important role in addressing this challenge. We demonstrate that topical vaginal administration of 3% G2-S16 prevents HIV-1 JR-CSF transmission in humanized (h)-BLT mice in 84% with no presence of HIV-1 RNA and vaginal lesions. Second-generation polyanionic carbosilane dendrimer G2-S16 with silica core and 16 sulfonate end-groups exerts anti-HIV-1 activity at an early stage of viral replication, blocking the gp120/CD4 interaction, acting on the virus, and inhibiting the cell-to-cell HIV-1 transmission, confirming its multifactorial and non-specific ability. This study represents the first demonstration that transmission of HIV-1 can be efficiently blocked by vaginally applied G2-S16 in h-BLT mice. These findings provide a step forward in the development of G2-S16-based vaginal microbicides to prevent vaginal HIV-1 transmission in humans. From the Clinical Editor HIV infections remain a significant problem worldwide and the major route of transmission is through sexual activity. In this article, the authors developed an antiviral agent containing polyanionic carbosilane dendrimer with silica core and 16 sulfonate end-groups. When applied vaginally, this was shown to exert anti-HIV protection. These positive findings may offer hope in the fight against the spread of HIV epidemic.
- Published
- 2015
46. Mechanosignaling through YAP and TAZ drives fibroblast activation and fibrosis
- Author
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Carol Feghali-Bostwick, Xaralabos Varelas, Laura E. Fredenburgh, Andrew M. Tager, Vladimir Vrbanac, Jeffrey C. Horowitz, Samantha E. Hiemer, Lauren E. Stopfer, Kyoung Moo Choi, Fei Liu, Daniel J. Tschumperlin, Clemens K. Probst, David Lagares, Aleksandar Marinkovic, Thomas H. Sisson, and Ivan O. Rosas
- Subjects
Male ,Pulmonary and Respiratory Medicine ,Physiology ,Pulmonary Fibrosis ,Cell Cycle Proteins ,WWTR1 ,Mechanotransduction, Cellular ,Extracellular matrix ,Mice ,Mice, Inbred NOD ,Transforming Growth Factor beta ,Fibrosis ,Physiology (medical) ,Plasminogen Activator Inhibitor 1 ,Serpin E2 ,medicine ,Animals ,Humans ,Mechanotransduction ,Fibroblast ,Lung ,Adaptor Proteins, Signal Transducing ,Hippo signaling pathway ,biology ,Intracellular Signaling Peptides and Proteins ,YAP-Signaling Proteins ,Cell Biology ,Transforming growth factor beta ,Fibroblasts ,Phosphoproteins ,medicine.disease ,Cell biology ,medicine.anatomical_structure ,Biochemistry ,Gene Knockdown Techniques ,Transcriptional Coactivator with PDZ-Binding Motif Proteins ,Mutation ,NIH 3T3 Cells ,Trans-Activators ,Call for Papers ,biology.protein ,Female ,Transcription Factors ,Transforming growth factor - Abstract
Pathological fibrosis is driven by a feedback loop in which the fibrotic extracellular matrix is both a cause and consequence of fibroblast activation. However, the molecular mechanisms underlying this process remain poorly understood. Here we identify yes-associated protein (YAP) (homolog of drosophila Yki) and transcriptional coactivator with PDZ-binding motif (TAZ) (also known as Wwtr1), transcriptional effectors of the Hippo pathway, as key matrix stiffness-regulated coordinators of fibroblast activation and matrix synthesis. YAP and TAZ are prominently expressed in fibrotic but not healthy lung tissue, with particularly pronounced nuclear expression of TAZ in spindle-shaped fibroblastic cells. In culture, both YAP and TAZ accumulate in the nuclei of fibroblasts grown on pathologically stiff matrices but not physiologically compliant matrices. Knockdown of YAP and TAZ together in vitro attenuates key fibroblast functions, including matrix synthesis, contraction, and proliferation, and does so exclusively on pathologically stiff matrices. Profibrotic effects of YAP and TAZ operate, in part, through their transcriptional target plasminogen activator inhibitor-1, which is regulated by matrix stiffness independent of transforming growth factor-β signaling. Immortalized fibroblasts conditionally expressing active YAP or TAZ mutant proteins overcome soft matrix limitations on growth and promote fibrosis when adoptively transferred to the murine lung, demonstrating the ability of fibroblast YAP/TAZ activation to drive a profibrotic response in vivo. Together, these results identify YAP and TAZ as mechanoactivated coordinators of the matrix-driven feedback loop that amplifies and sustains fibrosis.
- Published
- 2015
47. High sensitivity HPLC method for determination of the allysine concentration in tissue by use of a naphthol derivative
- Author
-
Andrew M. Tager, Chloe M. Jones, Peter Caravan, Philip A. Waghorn, and Bruno L. Oliveira
- Subjects
0301 basic medicine ,Analyte ,Swine ,Pulmonary Fibrosis ,Clinical Biochemistry ,Lysine ,Biochemistry ,High-performance liquid chromatography ,Article ,Analytical Chemistry ,03 medical and health sciences ,chemistry.chemical_compound ,Bleomycin ,Mice ,0302 clinical medicine ,Naphthalenesulfonates ,Fibrosis ,Limit of Detection ,medicine ,Animals ,Lung ,Aorta ,Chromatography, High Pressure Liquid ,Detection limit ,Chromatography ,biology ,Hydrolysis ,Reproducibility of Results ,Cell Biology ,General Medicine ,medicine.disease ,Mice, Inbred C57BL ,030104 developmental biology ,chemistry ,030220 oncology & carcinogenesis ,biology.protein ,Linear Models ,Acid hydrolysis ,Allysine ,Elastin ,2-Aminoadipic Acid - Abstract
Common to all fibrotic and metastatic diseases is the uncontrollable remodeling of tissue that leads to the accumulation of fibrous connective tissue components such as collagen and elastin. Build-up of fibrous tissue occurs through the cross-linking of collagen or elastin monomers, which is initiated through the oxidation of lysine residues to form α-aminoadipic-δ-semialdehyde (allysine). To provide a measure of the extent of collagen oxidation in disease models of fibrosis or metastasis, a rapid, sensitive HPLC method was developed to quantify the amount of allysine present in tissue. Allysine was reacted with sodium 2-naphthol-7-sulfonate under conditions typically applied for acid hydrolysis of tissues (6 M HCl, 110 °C, 24 h) to prepare AL-NP, a fluorescent bis-naphthol derivative of allysine. High performance liquid chromatography was applied for analysis of allysine content. Under optimal reaction and detection conditions, successful separation of AL-NP was achieved with excellent analytical performance attained. Good linear relationship (R 2 = 0.994) between peak area and concentration for AL-NP was attained for 0.35–175 pmol of analyte. A detection limit of 0.02 pmol in the standard sample with a 20 μL injection was achieved for AL-NP, with satisfactory recovery from 88 to 100% determined. The method was applied in the quantification of allysine in healthy and fibrotic mouse lung tissue, with the fibrotic tissue showing a 2.5 fold increase in the content of allysine.
- Published
- 2017
48. Molecular imaging of oxidized collagen quantifies pulmonary and hepatic fibrogenesis
- Author
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Bryan C. Fuchs, Peter Caravan, Daniel T. Schühle, Nicholas J. Rotile, Richard J. Looby, Michael Lanuti, Howard H. Chen, Clemens K. Probst, Lan Wei, Gregory Y. Lauwers, Justin Elliott, Luis F. Tapias, Kenneth K. Tanabe, Mari Mino-Kenudson, Philip A. Waghorn, Gaofeng Zhao, Chloe M. Jones, and Andrew M. Tager
- Subjects
0301 basic medicine ,medicine.medical_specialty ,Pathology ,medicine.diagnostic_test ,Chemistry ,Lysine ,Magnetic resonance imaging ,Lysyl oxidase ,General Medicine ,030204 cardiovascular system & hematology ,Hepatology ,medicine.disease ,3. Good health ,Extracellular matrix ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Fibrosis ,Internal medicine ,medicine ,Molecular imaging ,Hepatic fibrosis ,Research Article - Abstract
Fibrosis results from the dysregulation of tissue repair mechanisms affecting major organ systems, leading to chronic extracellular matrix buildup, and progressive, often fatal, organ failure. Current diagnosis relies on invasive biopsies. Noninvasive methods today cannot distinguish actively progressive fibrogenesis from stable scar, and thus are insensitive for monitoring disease activity or therapeutic responses. Collagen oxidation is a universal signature of active fibrogenesis that precedes collagen crosslinking. Biochemically targeting oxidized lysine residues formed by the action of lysyl oxidase on collagen with a small-molecule gadolinium chelate enables targeted molecular magnetic resonance imaging. This noninvasive direct biochemical elucidation of the fibrotic microenvironment specifically and robustly detected and staged pulmonary and hepatic fibrosis progression, and monitored therapeutic response in animal models. Furthermore, this paradigm is translatable and generally applicable to diverse fibroproliferative disorders.
- Published
- 2017
49. Molecular Magnetic Resonance Imaging of Lung Fibrogenesis with an Oxyamine-Based Probe
- Author
-
Nicholas J. Rotile, Howard H. Chen, Clemens K. Probst, Andrew M. Tager, Peter Caravan, Steffi K. Koerner, Philip A. Waghorn, Chloe M. Jones, and Diego dos Santos Ferreira
- Subjects
0301 basic medicine ,Pathology ,medicine.medical_specialty ,Gadolinium ,Pulmonary Fibrosis ,Molecular Conformation ,chemistry.chemical_element ,Lysyl oxidase ,01 natural sciences ,Catalysis ,Article ,Extracellular matrix ,03 medical and health sciences ,chemistry.chemical_compound ,Bleomycin ,Mice ,0302 clinical medicine ,Fibrosis ,medicine ,Animals ,Amines ,Chelating Agents ,Gadolinium-Chelate ,Lung ,medicine.diagnostic_test ,010405 organic chemistry ,Magnetic resonance imaging ,General Medicine ,General Chemistry ,medicine.disease ,Magnetic Resonance Imaging ,0104 chemical sciences ,030104 developmental biology ,medicine.anatomical_structure ,chemistry ,Molecular Probes ,030211 gastroenterology & hepatology ,Allysine ,2-Aminoadipic Acid - Abstract
Fibrogenesis is the active production of extracellular matrix in response to tissue injury. In many chronic diseases persistent fibrogenesis results in the accumulation of scar tissue, which can lead to organ failure and death. However, no non-invasive technique exists to assess this key biological process. All tissue fibrogenesis results in the formation of allysine, which enables collagen cross-linking and leads to tissue stiffening and scar formation. We report herein a novel allysine-binding gadolinium chelate (GdOA), that can non-invasively detect and quantify the extent of fibrogenesis using magnetic resonance imaging (MRI). We demonstrate that GdOA signal enhancement correlates with the extent of the disease and is sensitive to a therapeutic response.
- Published
- 2017
50. Uncoupling of the profibrotic and hemostatic effects of thrombin in lung fibrosis
- Author
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Clemens K. Probst, Barry S. Shea, Andrew M. Tager, Elizabeth M. Van Cott, Patricia L. Brazee, Katharine E. Black, Peter Caravan, Nicholas J. Rotile, Paul H. Weinreb, Francesco Blasi, David E. Sosnovik, and Shelia M. Violette
- Subjects
0301 basic medicine ,Lung ,biology ,business.industry ,General Medicine ,Lung injury ,medicine.disease ,Fibrin ,03 medical and health sciences ,Idiopathic pulmonary fibrosis ,030104 developmental biology ,Thrombin ,medicine.anatomical_structure ,Fibrosis ,Pulmonary fibrosis ,medicine ,biology.protein ,Cancer research ,business ,Research Article ,Discovery and development of direct thrombin inhibitors ,medicine.drug - Abstract
Fibrotic lung disease, most notably idiopathic pulmonary fibrosis (IPF), is thought to result from aberrant wound-healing responses to repetitive lung injury. Increased vascular permeability is a cardinal response to tissue injury, but whether it is mechanistically linked to lung fibrosis is unknown. We previously described a model in which exaggeration of vascular leak after lung injury shifts the outcome of wound-healing responses from normal repair to pathological fibrosis. Here we report that the fibrosis produced in this model is highly dependent on thrombin activity and its downstream signaling pathways. Direct thrombin inhibition with dabigatran significantly inhibited protease-activated receptor-1 (PAR1) activation, integrin αvβ6 induction, TGF-β activation, and the development of pulmonary fibrosis in this vascular leak-dependent model. We used a potentially novel imaging method - ultashort echo time (UTE) lung magnetic resonance imaging (MRI) with the gadolinium-based, fibrin-specific probe EP-2104R - to directly visualize fibrin accumulation in injured mouse lungs, and to correlate the antifibrotic effects of dabigatran with attenuation of fibrin deposition. We found that inhibition of the profibrotic effects of thrombin can be uncoupled from inhibition of hemostasis, as therapeutic anticoagulation with warfarin failed to downregulate the PAR1/αvβ6/TGF-β axis or significantly protect against fibrosis. These findings have direct and important clinical implications, given recent findings that warfarin treatment is not beneficial in IPF, and the clinical availability of direct thrombin inhibitors that our data suggest could benefit these patients.
- Published
- 2017
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